pCVD016 vector (V001565)

Basic Vector Information

      • Vector Name:
      • pCVD016
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 4632 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.

pCVD016 vector Vector Map

pCVD0164632 bp600120018002400300036004200M13 fwdEcoRVAgeIlpp T; lpp T; lpp terminatortrc promoterlac operatorattR1lac UV5 promoterCmRPccdB; upstream region of the ccdB ORF including promoter and RBS; derived from the destination cassette used in the Gateway Technology developed by Life TechnologiesccdBattR2rrnB T2 terminatorXbaIG5C5; GC-adaptor for seamless assembly, flanking functional devices, custom sequences or cyanobacterial replicons; 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 ℃.

pCVD016 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_49092        4632 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD016, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4632)
  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 4632)
  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 4632)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4632)
  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..4632
                     /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="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
                     /note="lpp T; lpp T; lpp terminator"
                     /regulatory_class="terminator"
     promoter        494..523
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    531..547
                     /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)."
     protein_bind    572..696
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        721..751
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             805..1461
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     regulatory      1501..1550
                     /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             1806..2108
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(2152..2276)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      2285..2312
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     misc_feature    2321..2326
                     /label=XbaI
                     /note="XbaI"
     misc_feature    2327..2347
                     /note="G5C5; GC-adaptor for seamless assembly, flanking 
                     functional devices, custom sequences or cyanobacterial 
                     replicons; synthetic"
     misc_feature    2345..2350
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    2369..2374
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(2411..2427)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2435..2451)
                     /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(2459..2489)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2504..2525)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2813..3401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3575..4432)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4433..4537)
                     /label=AmpR promoter
     misc_feature    4563..4568
                     /label=ZraI
                     /note="ZraI"

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