pCVD059 vector (V001528)

Basic Vector Information

      • Vector Name:
      • pCVD059
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 4989 bp
      • Type:
      • Broad host range vector
      • 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.

pCVD059 vector Vector Map

pCVD0594989 bp6001200180024003000360042004800M13 fwdEcoRVXbaIrrnB T2 terminatorattB2mEGFPccdBPccdB; upstream region of the ccdB ORF including promoter and RBS; derived from the destination cassette used in the Gateway Technology developed by Life TechnologiesCmRlac UV5 promoterC2G; 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 ℃.

pCVD059 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_49277        4989 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD059, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4989)
  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 4989)
  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 4989)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4989)
  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..4989
                     /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    complement(408..428)
                     /note="G5C5; GC-adaptor for seamless assembly, flanking 
                     functional devices, custom sequences or cyanobacterial 
                     replicons; synthetic"
     misc_feature    429..434
                     /label=XbaI
                     /note="XbaI"
     terminator      complement(442..469)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     protein_bind    478..502
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(522..1238)
                     /label=mEGFP
                     /note="enhanced GFP with monomerizing A206K mutation
                     (Zacharias et al., 2002)"
     CDS             complement(1285..1587)
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     regulatory      complement(1843..1892)
                     /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"
                     /regulatory_class="promoter"
     CDS             complement(1932..2588)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(2642..2672)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     misc_feature    complement(2684..2704)
                     /note="C2G; GC-adaptor for seamless assembly, flanking 
                     antibiotic resistance markers, functional devices or custom
                     sequences; synthetic"
     misc_feature    2702..2707
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    2726..2731
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(2768..2784)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2792..2808)
                     /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(2816..2846)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2861..2882)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3170..3758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3932..4789)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4790..4894)
                     /label=AmpR promoter
     misc_feature    4920..4925
                     /label=ZraI
                     /note="ZraI"

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