Basic Vector Information
pCVD050 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 ℃.
pCVD050 vector Sequence
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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