pCVD069 vector (V001521)

Basic Vector Information

Vector Name:
pCVD069
Antibiotic Resistance:
Kanamycin
Length:
3781 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.

pCVD069 vector Vector Map

pCVD0693781 bp60012001800240030003600M13 fwdEcoRVNheINeoR/KanRPnptII; upstream region of nptII ORF including promoter and RBS in INSD accession V00618; derived from pAM505AgeIC2G; GC-adaptor for seamless assembly, flanking antibiotic resistance markers, functional devices or custom sequences; syntheticXbaIM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoterZraI

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 ℃.

pCVD069 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_49312        3781 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD069, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3781)
  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 3781)
  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 3781)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3781)
  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..3781
                     /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=EcoRV
                     /note="EcoRV"
     misc_feature    408..428
                     /note="GC; GC-adaptor for seamless assembly, flanking
                     antibiotic resistance markers or Escherichia coli origins 
                     to be assembled with a cyanobacterial replicon; synthetic"
     misc_feature    429..434
                     /label=NheI
                     /note="NheI"
     CDS             complement(521..1312)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      complement(1313..1469)
                     /note="PnptII; upstream region of nptII ORF including
                     promoter and RBS in INSD accession V00618; derived from 
                     pAM505"
                     /regulatory_class="promoter"
     misc_feature    1470..1475
                     /label=AgeI
                     /note="AgeI"
     misc_feature    1476..1496
                     /note="C2G; GC-adaptor for seamless assembly, flanking 
                     antibiotic resistance markers, functional devices or custom
                     sequences; synthetic"
     misc_feature    1494..1499
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    1518..1523
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(1560..1576)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1584..1600)
                     /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(1608..1638)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1653..1674)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1962..2550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2724..3581)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3582..3686)
                     /label=AmpR promoter
     misc_feature    3712..3717
                     /label=ZraI
                     /note="ZraI"

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