Basic Vector Information
pCVD033 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 ℃.
pCVD033 vector Sequence
LOCUS 40924_49177 3334 bp DNA circular SYN 18-DEC-2018 DEFINITION Broad host range vector vector pCVD033, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3334) 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 3334) 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 3334) TITLE Direct Submission REFERENCE 4 (bases 1 to 3334) 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..3334 /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 389..394 /label=EcoRV /note="EcoRV" misc_feature 392..412 /note="C2G; GC-adaptor for seamless assembly, flanking antibiotic resistance markers, functional devices or custom sequences; synthetic" misc_feature 413..418 /label=AgeI /note="AgeI" promoter 425..453 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" regulatory 466..506 /note="RBS_LT; RBS and flanking sequences found in Life Tech pENTR-SD/D Topo, start downstream of the att site and includes the T7 gene enhancer, RBS, TOPO binding site and directional TOPO overhang; Life Technologies; synthetic" /regulatory_class="ribosome_binding_site" RBS 477..497 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 507..1223 /label=ECFP /note="enhanced CFP" protein_bind complement(1243..1267) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" terminator 1277..1304 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" misc_feature 1313..1318 /label=XbaI /note="XbaI" misc_feature 1319..1339 /note="G5C5; GC-adaptor for seamless assembly, flanking functional devices, custom sequences or cyanobacterial replicons; synthetic" misc_feature 1337..1342 /label=EcoRV /note="EcoRV" rep_origin complement(1587..2175) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2349..3206) /label=AmpR /note="beta-lactamase" promoter complement(3207..3311) /label=AmpR promoter
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