pCVD050 vector (V001537)

Basic Vector Information

      • Vector Name:
      • pCVD050
      • 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.

pCVD050 vector Vector Map

pCVD0504989 bp6001200180024003000360042004800M13 fwdEcoRVXbaIrepAPrepA; 682 nts upstream of repA-pDC1; derived from pSUN202; Nostoc sp. PCC8009SacICAP binding sitelac promoterlac operatorlacZ-alphaSacIC3G3; GC-adaptor for seamless assembly, flanking cyanobacterial replicons or Escherichia coli origins to be assembled with a cyanobacterial replicon; 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 ℃.

pCVD050 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_49232        4989 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD050, 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    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"
     CDS             complement(600..1634)
                     /codon_start=1
                     /product="repA"
                     /label=repA
                     /note="repA-pDC1; derived from pSUN202; Nostoc sp. PCC8009"
                     /protein_id="AII99670.1"
                     /translation="MANQNRAATQVEGLNHCPLTSQEDRSALSSNVPLPLVLILVKAEI
                     ILQPFMVRLHSGISRQPWARLWYLLRGFDDGSGRVKEIPLVLVEEILGVSQATVYRWLD
                     QGKAVGAFLSGYKVRIGLLTVYLGGLTKVAWHLNLKDWGVVAECPLWEVNANIRALTTG
                     IVTQRLQQRSRYAANRKLKPEYRKSYGAPHPNELLGDEGQSSFKLGAGEVPFVLHVSPS
                     RVFVSKSFIHYGTSQNAVSCKLGIHTRTVRRHQRTLGMEKRQLCQAKYEYAQLDRAVSN
                     EASECWAWSSEGTQSDIGYQSLGISHWVKGRSRFLMASCQEPESSSRTLTSCQRANCLG
                     VSSK"
     misc_feature    1221..1228
                     /label=SwaI
                     /note="SwaI"
     regulatory      complement(1635..2316)
                     /note="PrepA; 682 nts upstream of repA-pDC1; derived from 
                     pSUN202; Nostoc sp. PCC8009"
                     /regulatory_class="promoter"
     misc_feature    2113..2118
                     /label=MfeI
                     /note="MfeI"
     misc_feature    2317..2322
                     /label=SacI
                     /note="SacI"
     protein_bind    2336..2357
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2372..2402
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2410..2426
                     /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)."
     CDS             2447..2620
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
     misc_feature    2678..2683
                     /label=SacI
                     /note="SacI"
     misc_feature    2684..2704
                     /note="C3G3; GC-adaptor for seamless assembly, flanking 
                     cyanobacterial replicons or Escherichia coli origins to be 
                     assembled with a cyanobacterial replicon; 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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