Basic Vector Information
pCVD060 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 ℃.
pCVD060 vector Sequence
LOCUS 40924_49282 3982 bp DNA circular SYN 18-DEC-2018 DEFINITION Broad host range vector vector pCVD060, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3982) 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 3982) 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 3982) TITLE Direct Submission REFERENCE 4 (bases 1 to 3982) 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..3982 /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" misc_feature 516..523 /label=SwaI /note="SwaI" regulatory 560..609 /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 865..1167 /label=ccdB /note="CcdB, a bacterial toxin that poisons DNA gyrase" misc_feature 1211..1218 /label=SwaI /note="SwaI" misc_feature 1223..1228 /label=SacI /note="SacI" CDS 1238..1555 /label=miniSOG /note="mini singlet oxygen generator, a fluorescent flavoprotein suitable for correlated light and electron microscopy (Shu et al., 2011)" CDS 1586..1609 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" misc_feature 1620..1625 /label=SacI /note="SacI" terminator 1635..1662 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" misc_feature 1671..1676 /label=XbaI /note="XbaI" misc_feature 1677..1697 /note="G5C5; GC-adaptor for seamless assembly, flanking functional devices, custom sequences or cyanobacterial replicons; synthetic" misc_feature 1695..1700 /label=EcoRV /note="EcoRV" misc_feature 1719..1724 /label=XbaI /note="XbaI" primer_bind complement(1761..1777) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(1785..1801) /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(1809..1839) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(1854..1875) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(2163..2751) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2925..3782) /label=AmpR /note="beta-lactamase" promoter complement(3783..3887) /label=AmpR promoter misc_feature 3913..3918 /label=ZraI /note="ZraI"
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