Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B, Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW.
pCVD015 载体载体图谱
pCVD015 载体载体序列
LOCUS Exported 3585 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Broad host range vector vector pCVD015, complete sequence.
ACCESSION KM017870
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3585)
AUTHORS Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B,
Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW.
TITLE Broad-host-range vector system for synthetic biology and
biotechnology in cyanobacteria
JOURNAL Nucleic Acids Res. (2014) In press
PUBMED 25074377
REFERENCE 2 (bases 1 to 3585)
AUTHORS Taton A, Unglaub F, Wright N, Zeng WY, Paz Yepes J, Brahamsha B,
Palenik B, Peterson T, Haerizadeh F, Golden SS, Golden JW.
TITLE Direct Submission
JOURNAL Submitted (17-JUN-2014) Division of Biological Sciences, University
of California, San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA
REFERENCE 3 (bases 1 to 3585)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..3585
/organism="Broad host range vector vector pCVD015"
/mol_type="other DNA"
/note="ccdB-SwaI is a cloning device, harboring the ccdB
gene, flanked with SwaI sites and adaptor sequences for
seamless assembly reaction"
/db_xref="taxon:1531873"
misc_feature complement(146..404)
/note="lacZa-part2; lacZalpha fragment upstream of EcoICRI"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 405..410
/label=EcoRV
/note="EcoRV"
misc_feature 408..1300
/note="CVD-cassette; device for the GC-adaptor assembly:
expression/reporter cassette, promoter-reporter module,
custom sequence"
misc_feature 408..428
/note="C2G; GC-adaptor for seamless assembly, flanking
antibiotic resistance markers, functional devices or custom
sequences; synthetic"
misc_feature 429..434
/label=AgeI
/note="AgeI"
regulatory 454..482
/regulatory_class="terminator"
/note="lpp T; lpp T; lpp terminator"
misc_feature 516..523
/label=SwaI
/note="SwaI"
regulatory 560..609
/regulatory_class="promoter"
/note="PccdB; upstream region of the ccdB ORF including
promoter and RBS; derived from the destination cassette
used in the Gateway Technology developed by Life
Technologies"
CDS 865..1170
/codon_start=1
/product="ccdB"
/label=ccdB
/note="from a destination cassette used in the Gateway
Technology developed by Life Technologies"
/protein_id="AII99623.1"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
misc_feature 1211..1218
/label=SwaI
/note="SwaI"
misc_feature 1223..1228
/label=SacI
/note="SacI"
regulatory 1238..1266
/regulatory_class="terminator"
/note="rrnB T2; rrnB T2; rrnB T2 terminator partial
sequence"
terminator 1238..1265
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
misc_feature 1274..1279
/label=XbaI
/note="XbaI"
misc_feature 1280..1300
/note="G5C5; GC-adaptor for seamless assembly, flanking
functional devices, custom sequences or cyanobacterial
replicons; synthetic"
misc_feature 1298..1303
/label=EcoRV
/note="EcoRV"
misc_feature complement(1304..1368)
/note="lacZa-part1; lacZalpha fragment upstream of EcoICRI"
misc_feature 1322..1327
/label=XbaI
/note="XbaI"
primer_bind complement(1364..1380)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
regulatory complement(1369..1491)
/regulatory_class="promoter"
/note="PlacZa; lacI-lacZ intergenic region derived from
Escherichia coli including the promoter and RBS"
regulatory complement(1386..1406)
/regulatory_class="other"
/note="lacO; LacO; lac operator (lacO) from Invitrogen
pTrcHis2"
protein_bind 1388..1404
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(1412..1442)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1457..1478
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(1707..2380)
/direction=LEFT
/note="ori-pUC19; origin of replication of pUC19"
rep_origin complement(1766..2354)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2525..3385)
/codon_start=1
/product="bla"
/label=bla
/note="confers ampicillin resistance; beta-lactamase;
derived from pUC19"
/protein_id="AII99622.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3386..3490)
/gene="bla"
/label=AmpR promoter
regulatory complement(3386..3455)
/regulatory_class="promoter"
/note="Pbla; upstream region of the bla ORF including
promoter and RBS; derived from pUC19"
misc_feature 3516..3521
/label=ZraI
/note="ZraI"
Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B, Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW.
pCVD015 载体载体图谱
pCVD015 载体载体序列
LOCUS Exported 3585 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Broad host range vector vector pCVD015, complete sequence.
ACCESSION KM017870
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3585)
AUTHORS Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B,
Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW.
TITLE Broad-host-range vector system for synthetic biology and
biotechnology in cyanobacteria
JOURNAL Nucleic Acids Res. (2014) In press
PUBMED 25074377
REFERENCE 2 (bases 1 to 3585)
AUTHORS Taton A, Unglaub F, Wright N, Zeng WY, Paz Yepes J, Brahamsha B,
Palenik B, Peterson T, Haerizadeh F, Golden SS, Golden JW.
TITLE Direct Submission
JOURNAL Submitted (17-JUN-2014) Division of Biological Sciences, University
of California, San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA
REFERENCE 3 (bases 1 to 3585)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..3585
/organism="Broad host range vector vector pCVD015"
/mol_type="other DNA"
/note="ccdB-SwaI is a cloning device, harboring the ccdB
gene, flanked with SwaI sites and adaptor sequences for
seamless assembly reaction"
/db_xref="taxon:1531873"
misc_feature complement(146..404)
/note="lacZa-part2; lacZalpha fragment upstream of EcoICRI"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 405..410
/label=EcoRV
/note="EcoRV"
misc_feature 408..1300
/note="CVD-cassette; device for the GC-adaptor assembly:
expression/reporter cassette, promoter-reporter module,
custom sequence"
misc_feature 408..428
/note="C2G; GC-adaptor for seamless assembly, flanking
antibiotic resistance markers, functional devices or custom
sequences; synthetic"
misc_feature 429..434
/label=AgeI
/note="AgeI"
regulatory 454..482
/regulatory_class="terminator"
/note="lpp T; lpp T; lpp terminator"
misc_feature 516..523
/label=SwaI
/note="SwaI"
regulatory 560..609
/regulatory_class="promoter"
/note="PccdB; upstream region of the ccdB ORF including
promoter and RBS; derived from the destination cassette
used in the Gateway Technology developed by Life
Technologies"
CDS 865..1170
/codon_start=1
/product="ccdB"
/label=ccdB
/note="from a destination cassette used in the Gateway
Technology developed by Life Technologies"
/protein_id="AII99623.1"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
misc_feature 1211..1218
/label=SwaI
/note="SwaI"
misc_feature 1223..1228
/label=SacI
/note="SacI"
regulatory 1238..1266
/regulatory_class="terminator"
/note="rrnB T2; rrnB T2; rrnB T2 terminator partial
sequence"
terminator 1238..1265
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
misc_feature 1274..1279
/label=XbaI
/note="XbaI"
misc_feature 1280..1300
/note="G5C5; GC-adaptor for seamless assembly, flanking
functional devices, custom sequences or cyanobacterial
replicons; synthetic"
misc_feature 1298..1303
/label=EcoRV
/note="EcoRV"
misc_feature complement(1304..1368)
/note="lacZa-part1; lacZalpha fragment upstream of EcoICRI"
misc_feature 1322..1327
/label=XbaI
/note="XbaI"
primer_bind complement(1364..1380)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
regulatory complement(1369..1491)
/regulatory_class="promoter"
/note="PlacZa; lacI-lacZ intergenic region derived from
Escherichia coli including the promoter and RBS"
regulatory complement(1386..1406)
/regulatory_class="other"
/note="lacO; LacO; lac operator (lacO) from Invitrogen
pTrcHis2"
protein_bind 1388..1404
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(1412..1442)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1457..1478
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(1707..2380)
/direction=LEFT
/note="ori-pUC19; origin of replication of pUC19"
rep_origin complement(1766..2354)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2525..3385)
/codon_start=1
/product="bla"
/label=bla
/note="confers ampicillin resistance; beta-lactamase;
derived from pUC19"
/protein_id="AII99622.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3386..3490)
/gene="bla"
/label=AmpR promoter
regulatory complement(3386..3455)
/regulatory_class="promoter"
/note="Pbla; upstream region of the bla ORF including
promoter and RBS; derived from pUC19"
misc_feature 3516..3521
/label=ZraI
/note="ZraI"