//dna/chromosomalintegration
type: Collection
persistent identity: dna chromosomalintegration
display id: dna_chromosomalintegration
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
owned by: james
owned by: myers
member: Down
member: amyE 3 IS
member: K592200
member: BBa_K510000
member: BBa_K1930003
member: BBa_K1973006
member: K592202
member: Up
member: amyE 5 IS
member: BBa_K1930002

amyE 5 IS
type: ComponentDefinition
persistent identity: BBa K143001
display id: BBa_K143001
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: 5??? Integration Sequence for the amyE locus of B. subtilis
creator: Chris Hirst
created: 2008-08-26T11:00:00Z
modified: 2015-05-08T01:10:23Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _199_
m user id: 0
owner id: 3475
owning group id: 9
part status: Released HQ 2013
sample status: In stock
status: status/Available
bookmark: true
mutable description: The 5' integration sequence can be added to the front of a Biobrick construct and the 3' integration sequence specific for this locus (Part BBa_K143002) to the rear of the Biobrick construct to allow integration of the Biobrick construct into the chromosome of the gram positive bacterium B.subtilis. The AmyE locus was the first locus used for integration into ''B.subtilis'' by Shimotsu and Henner#1 and is still commonly used in vectors such as pDR111#2, pDL#3 and their derivatives. Integration at the AmyE locus removes the ability of ''B.subtilis'' to break down starch, which can be assayed with iodine as described by Cutting and Vander-horn#4. The 5' and 3' integration sequences for the AmyE locus were used to integrate the Imperial 2008 iGEM project primary construct into the ''B.sutbilis'' chromosome. <biblio> #1 pmid=3019840 #2 pmid=14597697 #3 ''Bacillus'' Genetic Stock Center [www.bgsc.org] #4 Cutting, S M.; Vander-Horn, P B. Genetic analysis. In: Harwood C R, Cutting S M. , editors. Molecular biological methods for Bacillus. Chichester, England: John Wiley & Sons, Ltd.; 1990. pp. 27???74. </biblio>
mutable notes: The AmyE integration sequence was taken from the vector after comparison by BLAST to the ''B.subtilis'' chromosome to identify the homologous sequences. The sequence present in both the host chromosome and the plasmid at the 5' end of the gene is the 5' sequence required for integration.
mutable provenance: The 5??? integration sequence was taken from the shuttle vector pDR111 which has been used in many studies on ''B.subtilis'', in particular in the studies of transcriptional control#1 #2 #3 <biblio> #1 pmid=14597697 #2 pmid=15937167 #3 pmid=12169614 </biblio>
owned by: james
owned by: myers
star: true
type: DnaRegion
role: partType/DNA
role: SO:0000110
sequence annotation: 5' AmyE homologous sequence
sequence: https://synbiohub.org/public/igem/BBa_K143001_sequence/1

5' AmyE homologous sequence
type: SequenceAnnotation
persistent identity: annotation1974145
display id: annotation1974145
version: 1
top level: amyE 5 IS
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K143001/annotation1974145/range1974145/1

https://synbiohub.org/public/igem/BBa_K143001/annotation1974145/range1974145/1
type: Range
persistent identity: range1974145
display id: range1974145
version: 1
direction: direction/forward
top level: amyE 5 IS
start: 1
end: 522
orientation: inline

K592200
type: ComponentDefinition
persistent identity: BBa K592200
display id: BBa_K592200
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Low to medium copy BioBrick standard vector
creator: Erik Gullberg, Antonio Ascue Avalos, Erik Lundin
created: 2011-09-20T11:00:00Z
modified: 2015-05-08T01:12:49Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _763_
m user id: 0
owner id: 10133
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: true
mutable description: BBa_K592200 is a BioBrick standard vector with low to medium copy p15A replication origin (BBa_I50032) and spectinomycin antibiotic resistance marker, usable for Lambda Red recombineering in E coli. The E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
mutable notes: To make the plasmid usable for Lambda Red recombineering, all sequences with homologies to the E coli chromosome had to be removed. To accomplish this, the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.
mutable provenance: Salmonella phage P22 late terminator.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000755
role: partType/Plasmid Backbone
sequence annotation: Terminator BBa_B0054
sequence annotation: Spectinomycin resistance marker
sequence annotation: T3 terminator BBa_B0055
sequence annotation: P22 late terminator
sequence annotation: VF2 site BBa_G00100
sequence annotation: Translational stop BBa_B0042
sequence annotation: SpcR promoter
sequence annotation: VR site BBa_G00102
sequence annotation: Terminator BBa_B0062
sequence annotation: Lambda red F primer
sequence annotation: p15A low to medium copy replication origin BBa_I50032
sequence annotation: Lambda Red R primer
sequence: https://synbiohub.org/public/igem/BBa_K592200_sequence/1

Terminator BBa_B0054
type: SequenceAnnotation
persistent identity: annotation2136370
display id: annotation2136370
version: 1
top level: K592200
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K592200/annotation2136370/range2136370/1

https://synbiohub.org/public/igem/BBa_K592200/annotation2136370/range2136370/1
type: Range
persistent identity: range2136370
display id: range2136370
version: 1
direction: direction/forward
top level: K592200
start: 47
end: 115
orientation: inline

Spectinomycin resistance marker
type: SequenceAnnotation
persistent identity: annotation2136375
display id: annotation2136375
version: 1
top level: K592200
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K592200/annotation2136375/range2136375/1

https://synbiohub.org/public/igem/BBa_K592200/annotation2136375/range2136375/1
type: Range
persistent identity: range2136375
display id: range2136375
version: 1
direction: direction/reverse
top level: K592200
start: 1841
end: 2840
orientation: inline

T3 terminator BBa_B0055
type: SequenceAnnotation
persistent identity: annotation2136379
display id: annotation2136379
version: 1
top level: K592200
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K592200/annotation2136379/range2136379/1

https://synbiohub.org/public/igem/BBa_K592200/annotation2136379/range2136379/1
type: Range
persistent identity: range2136379
display id: range2136379
version: 1
direction: direction/forward
top level: K592200
start: 3013
end: 3090
orientation: inline

P22 late terminator
type: SequenceAnnotation
persistent identity: annotation2136377
display id: annotation2136377
version: 1
top level: K592200
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K592200/annotation2136377/range2136377/1

https://synbiohub.org/public/igem/BBa_K592200/annotation2136377/range2136377/1
type: Range
persistent identity: range2136377
display id: range2136377
version: 1
direction: direction/forward
top level: K592200
start: 2952
end: 2992
orientation: inline

VF2 site BBa_G00100
type: SequenceAnnotation
persistent identity: annotation2136378
display id: annotation2136378
version: 1
top level: K592200
role: feature/primer binding
role: SO:0005850
location: https://synbiohub.org/public/igem/BBa_K592200/annotation2136378/range2136378/1

https://synbiohub.org/public/igem/BBa_K592200/annotation2136378/range2136378/1
type: Range
persistent identity: range2136378
display id: range2136378
version: 1
direction: direction/forward
top level: K592200
start: 2993
end: 3012
orientation: inline

Translational stop BBa_B0042
type: SequenceAnnotation
persistent identity: annotation2136380
display id: annotation2136380
version: 1
top level: K592200
role: SO:0000319
role: feature/stop
location: https://synbiohub.org/public/igem/BBa_K592200/annotation2136380/range2136380/1

https://synbiohub.org/public/igem/BBa_K592200/annotation2136380/range2136380/1
type: Range
persistent identity: range2136380
display id: range2136380
version: 1
direction: direction/forward
top level: K592200
start: 3091
end: 3102
orientation: inline

SpcR promoter
type: SequenceAnnotation
persistent identity: annotation2136376
display id: annotation2136376
version: 1
top level: K592200
role: SO:0000167
role: feature/promoter
location: https://synbiohub.org/public/igem/BBa_K592200/annotation2136376/range2136376/1

https://synbiohub.org/public/igem/BBa_K592200/annotation2136376/range2136376/1
type: Range
persistent identity: range2136376
display id: range2136376
version: 1
direction: direction/reverse
top level: K592200
start: 2907
end: 2933
orientation: inline

VR site BBa_G00102
type: SequenceAnnotation
persistent identity: annotation2136371
display id: annotation2136371
version: 1
top level: K592200
role: feature/primer binding
role: SO:0005850
location: https://synbiohub.org/public/igem/BBa_K592200/annotation2136371/range2136371/1

https://synbiohub.org/public/igem/BBa_K592200/annotation2136371/range2136371/1
type: Range
persistent identity: range2136371
display id: range2136371
version: 1
direction: direction/reverse
top level: K592200
start: 116
end: 135
orientation: inline

Terminator BBa_B0062
type: SequenceAnnotation
persistent identity: annotation2136372
display id: annotation2136372
version: 1
top level: K592200
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K592200/annotation2136372/range2136372/1

https://synbiohub.org/public/igem/BBa_K592200/annotation2136372/range2136372/1
type: Range
persistent identity: range2136372
display id: range2136372
version: 1
direction: direction/forward
top level: K592200
start: 136
end: 176
orientation: inline

Lambda red F primer
type: SequenceAnnotation
persistent identity: annotation2136374
display id: annotation2136374
version: 1
top level: K592200
role: feature/primer binding
role: SO:0005850
location: https://synbiohub.org/public/igem/BBa_K592200/annotation2136374/range2136374/1

https://synbiohub.org/public/igem/BBa_K592200/annotation2136374/range2136374/1
type: Range
persistent identity: range2136374
display id: range2136374
version: 1
direction: direction/forward
top level: K592200
start: 1726
end: 1745
orientation: inline

p15A low to medium copy replication origin BBa_I50032
type: SequenceAnnotation
persistent identity: annotation2136373
display id: annotation2136373
version: 1
top level: K592200
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K592200/annotation2136373/range2136373/1

https://synbiohub.org/public/igem/BBa_K592200/annotation2136373/range2136373/1
type: Range
persistent identity: range2136373
display id: range2136373
version: 1
direction: direction/forward
top level: K592200
start: 184
end: 1748
orientation: inline

Lambda Red R primer
type: SequenceAnnotation
persistent identity: annotation2136398
display id: annotation2136398
version: 1
top level: K592200
role: feature/primer binding
role: SO:0005850
location: https://synbiohub.org/public/igem/BBa_K592200/annotation2136398/range2136398/1

https://synbiohub.org/public/igem/BBa_K592200/annotation2136398/range2136398/1
type: Range
persistent identity: range2136398
display id: range2136398
version: 1
direction: direction/reverse
top level: K592200
start: 20
end: 41
orientation: inline

Translational stop BBa_B0042
type: SequenceAnnotation
persistent identity: annotation2136369
display id: annotation2136369
version: 1
top level: K592200
role: SO:0000319
role: feature/stop
location: https://synbiohub.org/public/igem/BBa_K592200/annotation2136369/range2136369/1

https://synbiohub.org/public/igem/BBa_K592200/annotation2136369/range2136369/1
type: Range
persistent identity: range2136369
display id: range2136369
version: 1
direction: direction/forward
top level: K592200
start: 35
end: 46
orientation: inline

amyE 3 IS
type: ComponentDefinition
persistent identity: BBa K143002
display id: BBa_K143002
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: 3??? Integration Sequence for the amyE locus of B. subtilis
creator: Chris Hirst
created: 2008-08-27T11:00:00Z
modified: 2015-05-08T01:10:23Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _199_
m user id: 0
owner id: 3475
owning group id: 9
part status: Released HQ 2013
sample status: In stock
status: status/Available
bookmark: true
mutable description: Integration sequences allow DNA to be incorporated into the chromosome of a host cell at a specific locus using leading (5') and trailing (3') DNA sequences that are the same as those at a specific locus of the chromosome. The 5' integration sequence can be added to the front of a Biobrick construct and the 3' integration sequence specific for this locus (Part BBa_K143002) to the rear of the Biobrick construct to allow integration of the Biobrick construct into the chromosome of the gram positive bacterium B.subtilis. The AmyE locus was the first locus used for integration into B.subtilis by Shimotsu and Henner[1] and is still commonly used in vectors such as pDR111[2], pDL[3] and their derivatives. Integration at the AmyE locus removes the ability of B.subtilis to break down starch, which can be assayed with iodine as described by Cutting and Vander-horn[4]. The 5' and 3' integration sequences for the AmyE locus were used to integrate the Imperial 2008 iGEM project primary construct into the B.sutbilis chromosome. References 1. Shimotsu H and Henner DJ. Construction of a single-copy integration vector and its use in analysis of regulation of the trp operon of Bacillus subtilis. Gene 1986; 43(1-2) 85-94. pmid:3019840 2.Nakano S, Kster-Schck E, Grossman AD, and Zuber P. Spx-dependent global transcriptional control is induced by thiol-specific oxidative stress in Bacillus subtilis. Proc Natl Acad Sci U S A 2003 Nov 11; 100(23) 13603-8. doi:10.1073/pnas.2235180100 pmid:14597697 3.Bacillus Genetic Stock Center; www.bgsc.org 4.Cutting, S M.; Vander-Horn, P B. Genetic analysis. In: Harwood C R, Cutting S M. , editors. Molecular biological methods for Bacillus. Chichester, England: John Wiley & Sons, Ltd.; 1990. pp. 27???74.
mutable notes: The AmyE integration sequence was taken from the vector after comparison by BLAST to the B.subtilis chromosome to identify the homologous sequences. The sequence present in both the host chromosome and the plasmid at the 3' end of the gene is the 3' sequence required for integration
mutable provenance: The 3??? integration sequence was taken from the shuttle vector pDR111 which has been used in many studies on B.subtilis, in particular in the studies of transcriptional control[1,2,3] References 1.Shimotsu H and Henner DJ. Construction of a single-copy integration vector and its use in analysis of regulation of the trp operon of Bacillus subtilis. Gene 1986; 43(1-2) 85-94. pmid:3019840. 2.Erwin KN, Nakano S, and Zuber P. Sulfate-dependent repression of genes that function in organosulfur metabolism in Bacillus subtilis requires Spx. J Bacteriol 2005 Jun; 187(12) 4042-9. doi:10.1128/JB.187.12.4042-4049.2005 pmid:15937167 3.Britton RA, Eichenberger P, Gonzalez-Pastor JE, Fawcett P, Monson R, Losick R, and Grossman AD. Genome-wide analysis of the stationary-phase sigma factor (sigma-H) regulon of Bacillus subtilis. J Bacteriol 2002 Sep; 184(17) 4881-90. pmid:12169614
owned by: james
owned by: myers
star: true
type: DnaRegion
role: partType/DNA
role: SO:0000110
sequence annotation: 3' AmyE homologous sequence
sequence: https://synbiohub.org/public/igem/BBa_K143002_sequence/1

3' AmyE homologous sequence
type: SequenceAnnotation
persistent identity: annotation1974146
display id: annotation1974146
version: 1
top level: amyE 3 IS
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K143002/annotation1974146/range1974146/1

https://synbiohub.org/public/igem/BBa_K143002/annotation1974146/range1974146/1
type: Range
persistent identity: range1974146
display id: range1974146
version: 1
direction: direction/forward
top level: amyE 3 IS
start: 1
end: 1005
orientation: inline

BBa_K1930002
type: ComponentDefinition
persistent identity: BBa K1930002
display id: BBa_K1930002
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Encryption key sequence in integration plasmid (BBa_K823023)
creator: Eike Mahlandt
created: 2016-10-15T11:00:00Z
modified: 2016-10-17T09:00:28Z
discontinued: true
dominant: false
experience: experience/Works
group u list: _2397_
m user id: 29748
owner id: 29748
owning group id: 9
status: status/Deleted
bookmark: false
mutable description: The iGEM team Groningen 2016 worked on bioencryption in order to safely store data in DNA. In our project we were mainly working with two sequences of DNA which were the encrypted message sequence and the encryption key sequence. The key sequence was created by our software. It is therefore artificial DNA. We ordered the encrypted key sequence as gBlock from IDT (Integrated DNA technologies). This part was created in the interest of having a genomic integration plasmid with our key sequence. We used an available BioBrick BBa_K823023 from igem Munich 2012. It is an integration plasmid for Bacillus subtilis, which can be used for cloning in E. coli as well. An RFP is inserted in BBa_K823023 for more efficient screening after transformation.
mutable notes: The iGEM team Groningen 2016 worked on bioencryption in order to safely store data in DNA. In our project we were mainly working with two sequences of DNA which were the encrypted message sequence and the encryption key sequence. The key sequence was created by our software. It is therefore artificial DNA. We ordered the encrypted key sequence as gBlock from IDT (Integrated DNA technologies). This part was created in the interest of having a genomic integration plasmid with our key sequence. We used an available BioBrick BBa_K823023 from igem Munich 2012. It is an integration plasmid for Bacillus subtilis, which can be used for cloning in E. coli as well. An RFP is inserted in BBa_K823023 for more efficient screening after transformation.
mutable provenance: Key sequence: Created by our CryptoGErM software. http://2016.igem.org/Team:Groningen/Encoding BBa_K823023: Created by iGEM Munich 2012. http://parts.igem.org/Part:BBa_K823023
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000155
role: partType/Plasmid
sequence annotation: bla
sequence annotation: amyE
sequence annotation: key sequence
sequence annotation: BBa_K1930000
sequence annotation: cat
sequence annotation: BBa_K823023
sequence: https://synbiohub.org/public/igem/BBa_K1930002_sequence/1

bla
type: SequenceAnnotation
persistent identity: annotation2514193
display id: annotation2514193
version: 1
top level: BBa_K1930002
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K1930002/annotation2514193/range2514193/1

https://synbiohub.org/public/igem/BBa_K1930002/annotation2514193/range2514193/1
type: Range
persistent identity: range2514193
display id: range2514193
version: 1
direction: direction/forward
top level: BBa_K1930002
start: 4340
end: 5200
orientation: inline

amyE
type: SequenceAnnotation
persistent identity: annotation2514192
display id: annotation2514192
version: 1
top level: BBa_K1930002
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1930002/annotation2514192/range2514192/1

https://synbiohub.org/public/igem/BBa_K1930002/annotation2514192/range2514192/1
type: Range
persistent identity: range2514192
display id: range2514192
version: 1
direction: direction/forward
top level: BBa_K1930002
start: 1358
end: 2386
orientation: inline

key sequence
type: SequenceAnnotation
persistent identity: annotation2514195
display id: annotation2514195
version: 1
top level: BBa_K1930002
role: SO:0000110
role: feature/dna
location: https://synbiohub.org/public/igem/BBa_K1930002/annotation2514195/range2514195/1

https://synbiohub.org/public/igem/BBa_K1930002/annotation2514195/range2514195/1
type: Range
persistent identity: range2514195
display id: range2514195
version: 1
direction: direction/forward
top level: BBa_K1930002
start: 6063
end: 6206
orientation: inline

BBa_K1930000
type: SequenceAnnotation
persistent identity: annotation2514196
display id: annotation2514196
version: 1
top level: BBa_K1930002
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K1930002/annotation2514196/range2514196/1

https://synbiohub.org/public/igem/BBa_K1930002/annotation2514196/range2514196/1
type: Range
persistent identity: range2514196
display id: range2514196
version: 1
direction: direction/forward
top level: BBa_K1930002
start: 6063
end: 6206
orientation: inline

cat
type: SequenceAnnotation
persistent identity: annotation2514191
display id: annotation2514191
version: 1
top level: BBa_K1930002
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K1930002/annotation2514191/range2514191/1

https://synbiohub.org/public/igem/BBa_K1930002/annotation2514191/range2514191/1
type: Range
persistent identity: range2514191
display id: range2514191
version: 1
direction: direction/forward
top level: BBa_K1930002
start: 337
end: 987
orientation: inline

BBa_K823023
type: SequenceAnnotation
persistent identity: annotation2514190
display id: annotation2514190
version: 1
top level: BBa_K1930002
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K1930002/annotation2514190/range2514190/1

https://synbiohub.org/public/igem/BBa_K1930002/annotation2514190/range2514190/1
type: Range
persistent identity: range2514190
display id: range2514190
version: 1
direction: direction/forward
top level: BBa_K1930002
start: 1
end: 6062
orientation: inline

amyE
type: SequenceAnnotation
persistent identity: annotation2514194
display id: annotation2514194
version: 1
top level: BBa_K1930002
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1930002/annotation2514194/range2514194/1

https://synbiohub.org/public/igem/BBa_K1930002/annotation2514194/range2514194/1
type: Range
persistent identity: range2514194
display id: range2514194
version: 1
direction: direction/forward
top level: BBa_K1930002
start: 5525
end: 6045
orientation: inline

Up
type: ComponentDefinition
persistent identity: BBa K1642005
display id: BBa_K1642005
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Upstream sequence from slr0168
creator: Can Yin
created: 2015-08-14T11:00:00Z
modified: 2015-09-16T07:27:55Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2059_
m user id: 24405
owner id: 24405
owning group id: 9
status: status/Planning
bookmark: false
mutable description: Slr
mutable notes: Slr
mutable provenance: Slr
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/DNA
role: SO:0000110
sequence: https://synbiohub.org/public/igem/BBa_K1642005_sequence/1

BBa_K1973006
type: ComponentDefinition
persistent identity: BBa K1973006
display id: BBa_K1973006
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: miniTn7BB-Gm-nahR-Psal-lapG
creator: Gabriel Ruiz Romero
created: 2016-10-12T11:00:00Z
modified: 2016-10-13T09:24:18Z
discontinued: false
dominant: false
group u list: _2440_
m user id: 29968
owner id: 29968
owning group id: 9
status: status/Planning
bookmark: false
mutable description: nahR (Pseudomonas putida, see BBa_K1031610) is expressed under the constitutive Pr promoter. It encodes a regulatory protein that activates the Psal promoter (see BBa_J61051) in the presence of salicylate. lapG (see BBa_K1973000) is expressed from the Psal promoter in the presence of this molecule. The miniTn7 tool allows the stable integration of this module in the chromosome of a wide range of bacteria (see http://parts.igem.org/Genome_Integration).
mutable notes: Designs are described in basic parts pages
mutable provenance: Basic parts were cloned into the miniTn7BB-Gm Backbone
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000804
role: partType/Composite
component: BBa_K510000
component: BBa_K1973033
component: BBa_K1973000
sequence annotation: BBa_K1973033
sequence annotation: BBa_K510000
sequence annotation: BBa_K1973000
sequence: https://synbiohub.org/public/igem/BBa_K1973006_sequence/1

BBa_K510000
type: Component
persistent identity: component2498468
display id: component2498468
version: 1
top level: BBa_K1973006
access: public
definition: BBa_K510000

BBa_K1973033
type: Component
persistent identity: component2498471
display id: component2498471
version: 1
top level: BBa_K1973006
access: public
definition: https://synbiohub.org/public/igem/BBa_K1973033/1

BBa_K1973000
type: Component
persistent identity: component2498475
display id: component2498475
version: 1
top level: BBa_K1973006
access: public
definition: https://synbiohub.org/public/igem/BBa_K1973000/1

BBa_K1973033
type: SequenceAnnotation
persistent identity: annotation2498471
display id: annotation2498471
version: 1
top level: BBa_K1973006
location: https://synbiohub.org/public/igem/BBa_K1973006/annotation2498471/range2498471/1
component: BBa_K1973033

https://synbiohub.org/public/igem/BBa_K1973006/annotation2498471/range2498471/1
type: Range
persistent identity: range2498471
display id: range2498471
version: 1
direction: direction/reverse
top level: BBa_K1973006
start: 4397
end: 5435
orientation: inline

BBa_K510000
type: SequenceAnnotation
persistent identity: annotation2498468
display id: annotation2498468
version: 1
top level: BBa_K1973006
location: https://synbiohub.org/public/igem/BBa_K1973006/annotation2498468/range2498468/1
component: BBa_K510000

https://synbiohub.org/public/igem/BBa_K1973006/annotation2498468/range2498468/1
type: Range
persistent identity: range2498468
display id: range2498468
version: 1
direction: direction/reverse
top level: BBa_K1973006
start: 1
end: 4388
orientation: inline

BBa_K1973000
type: SequenceAnnotation
persistent identity: annotation2498475
display id: annotation2498475
version: 1
top level: BBa_K1973006
location: https://synbiohub.org/public/igem/BBa_K1973006/annotation2498475/range2498475/1
component: BBa_K1973000

https://synbiohub.org/public/igem/BBa_K1973006/annotation2498475/range2498475/1
type: Range
persistent identity: range2498475
display id: range2498475
version: 1
direction: direction/reverse
top level: BBa_K1973006
start: 5444
end: 6106
orientation: inline

BBa_K1930003
type: ComponentDefinition
persistent identity: BBa K1930003
display id: BBa_K1930003
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Encrypted message sequence in integration plasmid (BBa_K823023)
creator: Barbora Waclawikova
created: 2016-10-15T11:00:00Z
modified: 2016-10-17T09:01:43Z
discontinued: true
dominant: false
experience: experience/Works
group u list: _2397_
m user id: 29748
owner id: 29748
owning group id: 9
status: status/Deleted
bookmark: false
mutable description: The iGEM team Groningen 2016 worked on bioencryption in order to safely store data in DNA. In our project we were mainly working with two sequences of DNA which were the encrypted message sequence and the encryption key sequence. The message sequence was created by our software. It is therefore artificial DNA. We ordered the encrypted message sequence as gBlock from IDT (Integrated DNA technologies). This part was created in the interest of having a genomic integration plasmid with our message sequence. We used an available BioBrick BBa_K823023 from igem Munich 2012. It is an integration plasmid for Bacillus subtilis, which can be used for cloning in E. coli as well. An RFP is inserted in BBa_K823023 for more efficient screening after transformation.
mutable notes: The iGEM team Groningen 2016 worked on bioencryption in order to safely store data in DNA. In our project we were mainly working with two sequences of DNA which were the encrypted message sequence and the encryption key sequence. The message sequence was created by our software. It is therefore artificial DNA. We ordered the encrypted message sequence as gBlock from IDT (Integrated DNA technologies). This part was created in the interest of having a genomic integration plasmid with our message sequence. We used an available BioBrick BBa_K823023 from igem Munich 2012. It is an integration plasmid for Bacillus subtilis, which can be used for cloning in E. coli as well. An RFP is inserted in BBa_K823023 for more efficient screening after transformation. For more information see plasmid construction under our lab journal: http://2016.igem.org/Team:Groningen/Experiments
mutable provenance: Message sequence: Created by our CryptoGErM software. http://2016.igem.org/Team:Groningen/Encoding BBa_K823023: Created by iGEM Munich 2012. http://parts.igem.org/Part:BBa_K823023
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000155
role: partType/Plasmid
sequence annotation: bla
sequence annotation: BBa_K823023
sequence annotation: BBa_K1930001
sequence annotation: cat
sequence annotation: message sequence
sequence annotation: amyE
sequence: https://synbiohub.org/public/igem/BBa_K1930003_sequence/1

bla
type: SequenceAnnotation
persistent identity: annotation2513448
display id: annotation2513448
version: 1
top level: BBa_K1930003
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K1930003/annotation2513448/range2513448/1

https://synbiohub.org/public/igem/BBa_K1930003/annotation2513448/range2513448/1
type: Range
persistent identity: range2513448
display id: range2513448
version: 1
direction: direction/forward
top level: BBa_K1930003
start: 4896
end: 5756
orientation: inline

BBa_K823023
type: SequenceAnnotation
persistent identity: annotation2513444
display id: annotation2513444
version: 1
top level: BBa_K1930003
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K1930003/annotation2513444/range2513444/1

https://synbiohub.org/public/igem/BBa_K1930003/annotation2513444/range2513444/1
type: Range
persistent identity: range2513444
display id: range2513444
version: 1
direction: direction/forward
top level: BBa_K1930003
start: 557
end: 6618
orientation: inline

BBa_K1930001
type: SequenceAnnotation
persistent identity: annotation2514197
display id: annotation2514197
version: 1
top level: BBa_K1930003
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K1930003/annotation2514197/range2514197/1

https://synbiohub.org/public/igem/BBa_K1930003/annotation2514197/range2514197/1
type: Range
persistent identity: range2514197
display id: range2514197
version: 1
direction: direction/forward
top level: BBa_K1930003
start: 1
end: 556
orientation: inline

cat
type: SequenceAnnotation
persistent identity: annotation2513446
display id: annotation2513446
version: 1
top level: BBa_K1930003
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K1930003/annotation2513446/range2513446/1

https://synbiohub.org/public/igem/BBa_K1930003/annotation2513446/range2513446/1
type: Range
persistent identity: range2513446
display id: range2513446
version: 1
direction: direction/forward
top level: BBa_K1930003
start: 893
end: 1543
orientation: inline

message sequence
type: SequenceAnnotation
persistent identity: annotation2513445
display id: annotation2513445
version: 1
top level: BBa_K1930003
role: SO:0000110
role: feature/dna
location: https://synbiohub.org/public/igem/BBa_K1930003/annotation2513445/range2513445/1

https://synbiohub.org/public/igem/BBa_K1930003/annotation2513445/range2513445/1
type: Range
persistent identity: range2513445
display id: range2513445
version: 1
direction: direction/forward
top level: BBa_K1930003
start: 1
end: 556
orientation: inline

amyE
type: SequenceAnnotation
persistent identity: annotation2513447
display id: annotation2513447
version: 1
top level: BBa_K1930003
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1930003/annotation2513447/range2513447/1

https://synbiohub.org/public/igem/BBa_K1930003/annotation2513447/range2513447/1
type: Range
persistent identity: range2513447
display id: range2513447
version: 1
direction: direction/forward
top level: BBa_K1930003
start: 1914
end: 2942
orientation: inline

amyE
type: SequenceAnnotation
persistent identity: annotation2513449
display id: annotation2513449
version: 1
top level: BBa_K1930003
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K1930003/annotation2513449/range2513449/1

https://synbiohub.org/public/igem/BBa_K1930003/annotation2513449/range2513449/1
type: Range
persistent identity: range2513449
display id: range2513449
version: 1
direction: direction/forward
top level: BBa_K1930003
start: 6081
end: 6601
orientation: inline

BBa_K510000
type: ComponentDefinition
persistent identity: BBa K510000
display id: BBa_K510000
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: pUC18Sfi-miniTn7BB-Gm
creator: David Caballero, Fernando Govantes
created: 2011-09-15T11:00:00Z
modified: 2015-05-08T01:12:30Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _673_
m user id: 0
owner id: 6157
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: true
mutable description: The miniTn7BB-Gm synthetic minitransposon was digested at the flanking SfiI sites and cloned into SfiI-digested pUC18-Sfi. This results in the elimination of the complete pUC18-SfiI multi-cloning site, except for the duplicated SfiI sites that remain on both sides of the transposon, thus facilitating its transfer to other vectors. This plasmid can be used to keep the mini-Tn7-Gm in E. coli and other enterics, and may be used for transposition into the genomes of non-enteric bacteria, in which it is non-replicative. The structure of the mini-Tn7BB-Gm transposon is shown in the figure below. Tn7R and Tn7L are the right and left ends of Tn7 transposon, respectively, required for recognition of the transposase machinery and transposition. FRT is the Flp recombinase target site for excision of the resistance once the transposon is inserted in the genome of a living organism. The gentamycin resistance cassette (Gm) is flanked by restriction sites in order to facilitate the generation of variants with different antibiotic resistance markers. The BioBrick cloning site (BCS) includes the prefix and suffix restrictions sites used in assembly standard 10, and it is the place where BioBricks are inserted for genome integration. The complete construction is flanked by SfiI restriction sites to facilitate cloning.
mutable notes: In order to make this vector compatible with assembly standard 10, several restriction sites were removed from the synthetic sequence.
mutable provenance: The pUC18-SfiI cloning vector was kindly provided by the Microbiology laboratory of Centro Andaluz de Biolog??a del Desarrollo (CABD) in Sevilla. The miniTn7BB-Gm synthetic transposon was designed by the iGEM 2011 team UPO-Sevilla based on the set of miniTn7 derivatives developed by Choi et al. (2005) and commercially synthesized by Mr. Gene (Regensburg, Germany). K.-H. Choi, J. B. Gaynor, K. G. White, C. Lopez, C. M. Bosio, R. R. Karkhoff-Schweizer & H. P. Schweizer (2005). A Tn7-based broad-range bacterial cloning and expression system. Nature Methods, vol.2 NO.6, June 2005, 443.
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000155
role: partType/Plasmid
sequence annotation: ColEI (pUC18) Replication origin
sequence annotation: Flipase recombination sequence (FRT)
sequence annotation: Prefix
sequence annotation: Terminator
sequence annotation: VR (BBa_G00101) primer binding site
sequence annotation: NcoI restrictino site
sequence annotation: SfiI restriction site
sequence annotation: Tn7-R end
sequence annotation: Suffix
sequence annotation: SphI restriction site
sequence annotation: E. coli his operon terminator
sequence annotation: Tn7-L end
sequence annotation: Gentamycin resistance
sequence annotation: NcoI restriction site
sequence annotation: double terminator (BBa_B0014)
sequence annotation: Ampicillin resistance (bla)
sequence annotation: VF2 (G00100) primer binding site
sequence: https://synbiohub.org/public/igem/BBa_K510000_sequence/1

ColEI (pUC18) Replication origin
type: SequenceAnnotation
persistent identity: annotation2137417
display id: annotation2137417
version: 1
top level: BBa_K510000
role: SO:0000110
role: feature/dna
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137417/range2137417/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137417/range2137417/1
type: Range
persistent identity: range2137417
display id: range2137417
version: 1
direction: direction/forward
top level: BBa_K510000
start: 1965
end: 2644
orientation: inline

Flipase recombination sequence (FRT)
type: SequenceAnnotation
persistent identity: annotation2137420
display id: annotation2137420
version: 1
top level: BBa_K510000
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137420/range2137420/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137420/range2137420/1
type: Range
persistent identity: range2137420
display id: range2137420
version: 1
direction: direction/forward
top level: BBa_K510000
start: 3329
end: 3376
orientation: inline

Prefix
type: SequenceAnnotation
persistent identity: annotation2137423
display id: annotation2137423
version: 1
top level: BBa_K510000
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137423/range2137423/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137423/range2137423/1
type: Range
persistent identity: range2137423
display id: range2137423
version: 1
direction: direction/forward
top level: BBa_K510000
start: 4367
end: 4387
orientation: inline

Terminator
type: SequenceAnnotation
persistent identity: annotation2137449
display id: annotation2137449
version: 1
top level: BBa_K510000
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137449/range2137449/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137449/range2137449/1
type: Range
persistent identity: range2137449
display id: range2137449
version: 1
direction: direction/forward
top level: BBa_K510000
start: 4326
end: 4359
orientation: inline

VR (BBa_G00101) primer binding site
type: SequenceAnnotation
persistent identity: annotation2137413
display id: annotation2137413
version: 1
top level: BBa_K510000
role: feature/primer binding
role: SO:0005850
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137413/range2137413/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137413/range2137413/1
type: Range
persistent identity: range2137413
display id: range2137413
version: 1
direction: direction/forward
top level: BBa_K510000
start: 157
end: 176
orientation: inline

NcoI restrictino site
type: SequenceAnnotation
persistent identity: annotation2137436
display id: annotation2137436
version: 1
top level: BBa_K510000
role: SO:0000110
role: feature/dna
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137436/range2137436/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137436/range2137436/1
type: Range
persistent identity: range2137436
display id: range2137436
version: 1
direction: direction/forward
top level: BBa_K510000
start: 3387
end: 3392
orientation: inline

SfiI restriction site
type: SequenceAnnotation
persistent identity: annotation2137418
display id: annotation2137418
version: 1
top level: BBa_K510000
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137418/range2137418/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137418/range2137418/1
type: Range
persistent identity: range2137418
display id: range2137418
version: 1
direction: direction/forward
top level: BBa_K510000
start: 3002
end: 3014
orientation: inline

Tn7-R end
type: SequenceAnnotation
persistent identity: annotation2137419
display id: annotation2137419
version: 1
top level: BBa_K510000
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137419/range2137419/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137419/range2137419/1
type: Range
persistent identity: range2137419
display id: range2137419
version: 1
direction: direction/forward
top level: BBa_K510000
start: 3014
end: 3213
orientation: inline

Suffix
type: SequenceAnnotation
persistent identity: annotation2137411
display id: annotation2137411
version: 1
top level: BBa_K510000
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137411/range2137411/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137411/range2137411/1
type: Range
persistent identity: range2137411
display id: range2137411
version: 1
direction: direction/forward
top level: BBa_K510000
start: 2
end: 21
orientation: inline

SphI restriction site
type: SequenceAnnotation
persistent identity: annotation2137425
display id: annotation2137425
version: 1
top level: BBa_K510000
role: SO:0000110
role: feature/dna
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137425/range2137425/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137425/range2137425/1
type: Range
persistent identity: range2137425
display id: range2137425
version: 1
direction: direction/forward
top level: BBa_K510000
start: 3381
end: 3386
orientation: inline

SphI restriction site
type: SequenceAnnotation
persistent identity: annotation2137440
display id: annotation2137440
version: 1
top level: BBa_K510000
role: SO:0000110
role: feature/dna
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137440/range2137440/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137440/range2137440/1
type: Range
persistent identity: range2137440
display id: range2137440
version: 1
direction: direction/forward
top level: BBa_K510000
start: 4190
end: 4195
orientation: inline

E. coli his operon terminator
type: SequenceAnnotation
persistent identity: annotation2137412
display id: annotation2137412
version: 1
top level: BBa_K510000
role: feature/stem loop
role: SO:0000313
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137412/range2137412/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137412/range2137412/1
type: Range
persistent identity: range2137412
display id: range2137412
version: 1
direction: direction/forward
top level: BBa_K510000
start: 22
end: 93
orientation: inline

SfiI restriction site
type: SequenceAnnotation
persistent identity: annotation2137415
display id: annotation2137415
version: 1
top level: BBa_K510000
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137415/range2137415/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137415/range2137415/1
type: Range
persistent identity: range2137415
display id: range2137415
version: 1
direction: direction/forward
top level: BBa_K510000
start: 348
end: 360
orientation: inline

Flipase recombination sequence (FRT)
type: SequenceAnnotation
persistent identity: annotation2137422
display id: annotation2137422
version: 1
top level: BBa_K510000
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137422/range2137422/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137422/range2137422/1
type: Range
persistent identity: range2137422
display id: range2137422
version: 1
direction: direction/forward
top level: BBa_K510000
start: 4196
end: 4243
orientation: inline

Tn7-L end
type: SequenceAnnotation
persistent identity: annotation2137414
display id: annotation2137414
version: 1
top level: BBa_K510000
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137414/range2137414/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137414/range2137414/1
type: Range
persistent identity: range2137414
display id: range2137414
version: 1
direction: direction/forward
top level: BBa_K510000
start: 182
end: 347
orientation: inline

Gentamycin resistance
type: SequenceAnnotation
persistent identity: annotation2137421
display id: annotation2137421
version: 1
top level: BBa_K510000
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137421/range2137421/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137421/range2137421/1
type: Range
persistent identity: range2137421
display id: range2137421
version: 1
direction: direction/reverse
top level: BBa_K510000
start: 3393
end: 3926
orientation: inline

NcoI restriction site
type: SequenceAnnotation
persistent identity: annotation2137439
display id: annotation2137439
version: 1
top level: BBa_K510000
role: SO:0000110
role: feature/dna
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137439/range2137439/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137439/range2137439/1
type: Range
persistent identity: range2137439
display id: range2137439
version: 1
direction: direction/forward
top level: BBa_K510000
start: 4184
end: 4189
orientation: inline

double terminator (BBa_B0014)
type: SequenceAnnotation
persistent identity: annotation2137424
display id: annotation2137424
version: 1
top level: BBa_K510000
role: SO:0000804
role: feature/BioBrick
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137424/range2137424/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137424/range2137424/1
type: Range
persistent identity: range2137424
display id: range2137424
version: 1
direction: direction/forward
top level: BBa_K510000
start: 3224
end: 3318
orientation: inline

Ampicillin resistance (bla)
type: SequenceAnnotation
persistent identity: annotation2137416
display id: annotation2137416
version: 1
top level: BBa_K510000
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137416/range2137416/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137416/range2137416/1
type: Range
persistent identity: range2137416
display id: range2137416
version: 1
direction: direction/forward
top level: BBa_K510000
start: 966
end: 1826
orientation: inline

VF2 (G00100) primer binding site
type: SequenceAnnotation
persistent identity: annotation2137442
display id: annotation2137442
version: 1
top level: BBa_K510000
role: feature/primer binding
role: SO:0005850
location: https://synbiohub.org/public/igem/BBa_K510000/annotation2137442/range2137442/1

https://synbiohub.org/public/igem/BBa_K510000/annotation2137442/range2137442/1
type: Range
persistent identity: range2137442
display id: range2137442
version: 1
direction: direction/forward
top level: BBa_K510000
start: 4249
end: 4268
orientation: inline

K592202
type: ComponentDefinition
persistent identity: BBa K592202
display id: BBa_K592202
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Low to medium copy Lambda Red recombineering compatible plasmid
creator: Antonio Ascue Avalos
created: 2011-09-20T11:00:00Z
modified: 2015-05-08T01:12:49Z
discontinued: false
dominant: false
experience: experience/Works
group u list: _763_
m user id: 0
owner id: 10133
owning group id: 9
sample status: It's complicated
status: status/Planning
bookmark: false
mutable description: BBa_K592202 is a BioBrick friendly vector with low to medium copy p15A replication origin (BBa_I50032) and a kanamycin antibiotic resistance marker, usable for Lambda Red recombineering in E coli. The E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22, and the kanamycin resistance marker is flanked by FRT sites so that the resistance marker can be removed after chromosomal integration using the FLP recombinase.
mutable notes: To make the plasmid usable for Lambda Red recombineering, all sequences with homologies to the E coli chromosome had to be removed. To accomplish this, the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22. To be able to remove the antibiotic resistance marker after chromosomal integration, the kanamycin marker from the pKD4 plasmid (Datsenko and Wanner, 2000) was used. The resistance marker can be removed after chromosomal integration using the FLP recombinase, leaving only the plasmid insert and a scar behind.
mutable provenance: The P22 late terminator come from the genome of the Salmonella phage P22 (GenBank: AF217253.1, 19662..19702), and the kanamycin resistance marker flanked by FRT sites comes from the pKD4 plasmid (Datsenko and Wanner, 2000).
owned by: james
owned by: myers
star: false
type: DnaRegion
role: SO:0000755
role: partType/Plasmid Backbone
sequence annotation: L-red_F_primer
sequence annotation: VF2 site BBa_G00100
sequence annotation: FRT site
sequence annotation: Translational stop BBa_B0042
sequence annotation: Terminator BBa_B0062
sequence annotation: L-red_R_primer
sequence annotation: VR site BBa_G00102
sequence annotation: P22 late terminator
sequence annotation: Kanamycin resistance marker
sequence annotation: T3 terminator BBa_B0055
sequence annotation: p15A ori BBa_I50032
sequence annotation: Terminator BBa_B0054
sequence: https://synbiohub.org/public/igem/BBa_K592202_sequence/1

L-red_F_primer
type: SequenceAnnotation
persistent identity: annotation2137824
display id: annotation2137824
version: 1
top level: K592202
role: feature/primer binding
role: SO:0005850
location: https://synbiohub.org/public/igem/BBa_K592202/annotation2137824/range2137824/1

https://synbiohub.org/public/igem/BBa_K592202/annotation2137824/range2137824/1
type: Range
persistent identity: range2137824
display id: range2137824
version: 1
direction: direction/forward
top level: K592202
start: 1726
end: 1745
orientation: inline

VF2 site BBa_G00100
type: SequenceAnnotation
persistent identity: annotation2137829
display id: annotation2137829
version: 1
top level: K592202
role: feature/primer binding
role: SO:0005850
location: https://synbiohub.org/public/igem/BBa_K592202/annotation2137829/range2137829/1

https://synbiohub.org/public/igem/BBa_K592202/annotation2137829/range2137829/1
type: Range
persistent identity: range2137829
display id: range2137829
version: 1
direction: direction/forward
top level: K592202
start: 3286
end: 3305
orientation: inline

FRT site
type: SequenceAnnotation
persistent identity: annotation2137827
display id: annotation2137827
version: 1
top level: K592202
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K592202/annotation2137827/range2137827/1

https://synbiohub.org/public/igem/BBa_K592202/annotation2137827/range2137827/1
type: Range
persistent identity: range2137827
display id: range2137827
version: 1
direction: direction/forward
top level: K592202
start: 3172
end: 3219
orientation: inline

Translational stop BBa_B0042
type: SequenceAnnotation
persistent identity: annotation2137804
display id: annotation2137804
version: 1
top level: K592202
role: SO:0000319
role: feature/stop
location: https://synbiohub.org/public/igem/BBa_K592202/annotation2137804/range2137804/1

https://synbiohub.org/public/igem/BBa_K592202/annotation2137804/range2137804/1
type: Range
persistent identity: range2137804
display id: range2137804
version: 1
direction: direction/forward
top level: K592202
start: 35
end: 46
orientation: inline

Terminator BBa_B0062
type: SequenceAnnotation
persistent identity: annotation2137822
display id: annotation2137822
version: 1
top level: K592202
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K592202/annotation2137822/range2137822/1

https://synbiohub.org/public/igem/BBa_K592202/annotation2137822/range2137822/1
type: Range
persistent identity: range2137822
display id: range2137822
version: 1
direction: direction/forward
top level: K592202
start: 136
end: 176
orientation: inline

Translational stop BBa_B0042
type: SequenceAnnotation
persistent identity: annotation2137846
display id: annotation2137846
version: 1
top level: K592202
role: SO:0000319
role: feature/stop
location: https://synbiohub.org/public/igem/BBa_K592202/annotation2137846/range2137846/1

https://synbiohub.org/public/igem/BBa_K592202/annotation2137846/range2137846/1
type: Range
persistent identity: range2137846
display id: range2137846
version: 1
direction: direction/forward
top level: K592202
start: 3384
end: 3395
orientation: inline

L-red_R_primer
type: SequenceAnnotation
persistent identity: annotation2137792
display id: annotation2137792
version: 1
top level: K592202
role: feature/primer binding
role: SO:0005850
location: https://synbiohub.org/public/igem/BBa_K592202/annotation2137792/range2137792/1

https://synbiohub.org/public/igem/BBa_K592202/annotation2137792/range2137792/1
type: Range
persistent identity: range2137792
display id: range2137792
version: 1
direction: direction/reverse
top level: K592202
start: 20
end: 41
orientation: inline

VR site BBa_G00102
type: SequenceAnnotation
persistent identity: annotation2137821
display id: annotation2137821
version: 1
top level: K592202
role: feature/primer binding
role: SO:0005850
location: https://synbiohub.org/public/igem/BBa_K592202/annotation2137821/range2137821/1

https://synbiohub.org/public/igem/BBa_K592202/annotation2137821/range2137821/1
type: Range
persistent identity: range2137821
display id: range2137821
version: 1
direction: direction/reverse
top level: K592202
start: 116
end: 135
orientation: inline

P22 late terminator
type: SequenceAnnotation
persistent identity: annotation2137828
display id: annotation2137828
version: 1
top level: K592202
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K592202/annotation2137828/range2137828/1

https://synbiohub.org/public/igem/BBa_K592202/annotation2137828/range2137828/1
type: Range
persistent identity: range2137828
display id: range2137828
version: 1
direction: direction/forward
top level: K592202
start: 3245
end: 3285
orientation: inline

Kanamycin resistance marker
type: SequenceAnnotation
persistent identity: annotation2137826
display id: annotation2137826
version: 1
top level: K592202
role: feature/cds
role: SO:0000316
location: https://synbiohub.org/public/igem/BBa_K592202/annotation2137826/range2137826/1

https://synbiohub.org/public/igem/BBa_K592202/annotation2137826/range2137826/1
type: Range
persistent identity: range2137826
display id: range2137826
version: 1
direction: direction/reverse
top level: K592202
start: 2017
end: 2811
orientation: inline

FRT site
type: SequenceAnnotation
persistent identity: annotation2137825
display id: annotation2137825
version: 1
top level: K592202
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K592202/annotation2137825/range2137825/1

https://synbiohub.org/public/igem/BBa_K592202/annotation2137825/range2137825/1
type: Range
persistent identity: range2137825
display id: range2137825
version: 1
direction: direction/forward
top level: K592202
start: 1781
end: 1826
orientation: inline

T3 terminator BBa_B0055
type: SequenceAnnotation
persistent identity: annotation2137830
display id: annotation2137830
version: 1
top level: K592202
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K592202/annotation2137830/range2137830/1

https://synbiohub.org/public/igem/BBa_K592202/annotation2137830/range2137830/1
type: Range
persistent identity: range2137830
display id: range2137830
version: 1
direction: direction/forward
top level: K592202
start: 3306
end: 3383
orientation: inline

p15A ori BBa_I50032
type: SequenceAnnotation
persistent identity: annotation2137823
display id: annotation2137823
version: 1
top level: K592202
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K592202/annotation2137823/range2137823/1

https://synbiohub.org/public/igem/BBa_K592202/annotation2137823/range2137823/1
type: Range
persistent identity: range2137823
display id: range2137823
version: 1
direction: direction/forward
top level: K592202
start: 184
end: 1748
orientation: inline

Terminator BBa_B0054
type: SequenceAnnotation
persistent identity: annotation2137820
display id: annotation2137820
version: 1
top level: K592202
role: SO:0000110
role: feature/misc
location: https://synbiohub.org/public/igem/BBa_K592202/annotation2137820/range2137820/1

https://synbiohub.org/public/igem/BBa_K592202/annotation2137820/range2137820/1
type: Range
persistent identity: range2137820
display id: range2137820
version: 1
direction: direction/forward
top level: K592202
start: 47
end: 115
orientation: inline

Down
type: ComponentDefinition
persistent identity: BBa K1642006
display id: BBa_K1642006
version: 1
was generated by: https://synbiohub.org/public/igem/igem2sbol/1
description: Downstream sequence from slr0168
creator: Can Yin
created: 2015-08-14T11:00:00Z
modified: 2015-09-16T01:32:30Z
discontinued: false
dominant: false
experience: experience/None
group u list: _2059_
m user id: 24423
owner id: 24405
owning group id: 9
status: status/Planning
bookmark: false
mutable description: Slr
mutable notes: Slr
mutable provenance: Slr
owned by: james
owned by: myers
star: false
type: DnaRegion
role: partType/DNA
role: SO:0000110
sequence: https://synbiohub.org/public/igem/BBa_K1642006_sequence/1