Basic Vector Information
pCVD012 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 ℃.
pCVD012 vector Sequence
LOCUS 40924_49072 3400 bp DNA circular SYN 18-DEC-2018 DEFINITION Broad host range vector vector pCVD012, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3400) 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 3400) 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 3400) TITLE Direct Submission REFERENCE 4 (bases 1 to 3400) 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..3400 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 293..309 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 320..338 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 370..375 /label=EcoRV /note="EcoRV" misc_feature complement(373..1425) /note="CVD-ANTR; device for the GC-adaptor assembly: antibiotic resistance markers" misc_feature complement(373..393) /note="C2G; GC-adaptor for seamless assembly, flanking antibiotic resistance markers, functional devices or custom sequences; synthetic" misc_feature 394..399 /label=AgeI /note="AgeI" regulatory 406..562 /note="PnptII; upstream region of nptII ORF including promoter and RBS in INSD accession V00618; derived from pAM505" /regulatory_class="promoter" CDS 563..1354 /codon_start=1 /product="nptII_A7120" /label=nptII_A7120 /note="confers kanamycin resistance; aminoglycoside phosphotransferase" /protein_id="AII99780.1" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" regulatory 1363..1389 /note="trpA T; trpA terminator from Lorist6 INSD accession X98450, one nt shorter than reference; Lorist6" /regulatory_class="terminator" misc_feature 1399..1404 /label=NheI /note="NheI" misc_feature complement(1405..1425) /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 1423..1428 /label=EcoRV /note="EcoRV" rep_origin complement(1653..2241) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2415..3272) /label=AmpR /note="beta-lactamase" promoter complement(3273..3377) /label=AmpR promoter
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