Basic Vector Information
- Vector Name:
- pCVD052
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6852 bp
- Type:
- Broad host range vector
- Replication origin:
- ori
- Source/Author:
- Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B, Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW.
pCVD052 vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pCVD052 vector Sequence
LOCUS 40924_49242 6852 bp DNA circular SYN 18-DEC-2018
DEFINITION Broad host range vector vector pCVD052, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6852)
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 6852)
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 6852)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6852)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res. (2014) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(17-JUN-2014) Division of Biological Sciences, University of
California, San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6852
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 405..410
/label=ZraI
/note="ZraI"
misc_feature complement(408..428)
/note="C3G3; GC-adaptor for seamless assembly, flanking
cyanobacterial replicons; synthetic"
misc_feature 429..434
/label=SacI
/note="SacI"
CDS complement(492..665)
/label=lacZ-alpha
/note="LacZ-alpha fragment of beta-galactosidase"
regulatory complement(667..789)
/note="PlacZa; lacI-lacZ intergenic region derived from
Escherichia coli including the promoter and RBS"
/regulatory_class="promoter"
regulatory complement(684..704)
/note="lacO; LacO; lac operator (lacO) from Invitrogen
pTrcHis2"
/regulatory_class="other"
protein_bind 686..702
/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(710..740)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 755..776
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
misc_feature 790..795
/label=SacI
/note="SacI"
regulatory 796..1445
/note="PrepA; PrepA; upstream region of repA of pDU1
including promoter and ribosomal binding site,derived from
pAM505; Nostoc sp. PCC7524"
/regulatory_class="promoter"
misc_feature 1195..1200
/label=MfeI
/note="MfeI"
CDS 1446..2567
/codon_start=1
/product="repA"
/label=repA
/note="repA-pDU1; derived from pAM0505; Nostoc sp. PCC7524"
/protein_id="AII99679.1"
/translation="MISPESLNTNVSLTLLTRELQTEIILGTYTVRVHTRIGREPCARL
WYLCRALDKDGSGHLTLPLPVVQTFLDCSDKSVYRWLQDGKKIGAFRRYKIKAGMITVY
LGGMFQVCYNLNLKRWGDVAVVPLVQVLSDLRSLTTGIVTQSFQQKSRYAANRQLKPEY
RKLFGAPHPNELVKDTRQSSLKSPEGEVPCVLHISSSRIFVSKSFIHYGTSQKAVSCEL
GIHKRTVRRHQKQLGMNRRQLCQAKIEYNQLRHARNNDASEFWAFTGTKTDIGYQVMGD
AVVFSDGIAPGAKKRQPNTYQIDATEFDGRLFKVGDKVFMNRCNIYREQFTLTTMSAAR
RKYHFKLSQCHFSENRAGRVGNRFVIGCHSGEI"
misc_feature 2262..2267
/label=EcoRV
/note="EcoRV"
regulatory 2646..2673
/note="rho-ITTS; rho-ITTS; rho-independent transcriptional
termination signal, downstream of repA in pDU1, from
pAM505; Nostoc sp. PCC7524"
/regulatory_class="terminator"
misc_feature 3084..3089
/label=XbaI
/note="XbaI"
CDS complement(3188..3739)
/codon_start=1
/product="orfC"
/label=orfC
/note="orfC-pDU1; derived from pAM0505;
integrase/recombinase plasmid stability; similar to INSD
accession AAM76170; Nostoc sp. PCC7524"
/protein_id="AII99678.1"
/translation="MKVNGNGRAKILTSDELRRLFSDGFTTPRDRVLFGICLFTGCRVS
EALALQTTDIKGETLTFRKSTTKGKLKTRVVDIQPGLAALMADYHPKPGTLFPGMRGVS
DRLTRYAADKILRDAAKRIGLEGISTHSFRRTALNQMSSAGIPLRHIQEISGHNDLGTL
QRYLEVTPEQRRKAVSVIGF"
misc_feature 3276..3281
/label=EcoRV
/note="EcoRV"
misc_feature 3617..3622
/label=AgeI
/note="AgeI"
misc_feature 4044..4049
/label=XbaI
/note="XbaI"
CDS complement(4111..4464)
/codon_start=1
/product="hyp1"
/label=hyp1
/note="hypothetical protein; derived from pAM505; Nostoc
sp. PCC7524"
/protein_id="AII99677.1"
/translation="MADKTLATFRIDSEEWESFKNLASSESSNASALLTEFVRWYLAGN
RFNTPTSHTPTHLDTSLEQRIDNIEQRLDKVTTNNLDNIDEFIDKRIEDNLATRLDKLQ
SQLEELRGKSKAR"
misc_feature 4163..4168
/label=XbaI
/note="XbaI"
misc_feature 4220..4225
/label=XbaI
/note="XbaI"
misc_feature 4244..4249
/label=XbaI
/note="XbaI"
misc_feature 4289..4294
/label=XbaI
/note="XbaI"
misc_feature 4533..4538
/label=XbaI
/note="XbaI"
misc_feature 4537..4542
/label=EcoRV
/note="EcoRV"
misc_feature 4541..4546
/label=XbaI
/note="XbaI"
misc_feature complement(4547..4567)
/note="G5C5; GC-adaptor for seamless assembly, flanking a
BHR replicons, neutral sites, or cyanobacterial replicons;
synthetic"
misc_feature 4565..4570
/label=ZraI
/note="ZraI"
misc_feature 4589..4594
/label=XbaI
/note="XbaI"
primer_bind complement(4631..4647)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4655..4671)
/label=lac operator
/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(4679..4709)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4724..4745)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5033..5621)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5795..6652)
/label=AmpR
/note="beta-lactamase"
promoter complement(6653..6757)
/label=AmpR promoter
misc_feature 6783..6788
/label=ZraI
/note="ZraI"
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