Basic Vector Information
pVCC 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 ℃.
pVCC vector Sequence
LOCUS 40924_45908 5155 bp DNA circular SYN 18-DEC-2018 DEFINITION Transformation and cloning vector pVCC, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5155) AUTHORS Vannini A, Agriesti F, Mosca F, Roncarati D, Scarlato V, Danielli A. TITLE A convenient and robust in vivo reporter system to monitor gene expression in the human pathogen Helicobacter pylori JOURNAL Appl. Environ. Microbiol. 78 (18), 6524-6533 (2012) PUBMED 22773640 REFERENCE 2 (bases 1 to 5155) AUTHORS Danielli A. TITLE Direct Submission JOURNAL Submitted (01-SEP-2010) Department of Biology, University of Bologna, via Selmi, 3, Bologna, BO 40126, Italy REFERENCE 3 (bases 1 to 5155) TITLE Direct Submission REFERENCE 4 (bases 1 to 5155) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl. Environ. Microbiol."; date: "2012"; volume: "78"; issue: "18"; pages: "6524-6533" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (01-SEP-2010) Department of Biology, University of Bologna, via Selmi, 3, Bologna, BO 40126, Italy" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..5155 /mol_type="other DNA" /organism="synthetic DNA construct" misc_recomb 1..564 /note="flanking region for double homologous recombination in Helicobacter pylori G27 vac::aphA-3/luxCDABE acceptor strain" CDS 27..542 /codon_start=1 /gene="cysS" /product="CysS" /label=cysS /protein_id="ADZ38982.1" /translation="YLLGVHYRSVLNFNEEDLLVSKKRLDKIYRLKQRVLGTLGGINPN FKKEILECMQDDLNVSKALSVLESMLSSTNEKLDQNPKNKALKGEILANLKFIEELLGI GFKDPSAYFQLGVSESEKQEIENKIEERKRAKEQKNFLKADSIREELLKQKIALMDTPQ GTIWEKFF" gene 27..542 /gene="cysS" /label=cysS /note="Helicobacter pylori cysS cistron 5' end" CDS complement(683..1303) /note="Chloramphenicol acetyltransferase from Campylobacter coli. Accession#: P22782" misc_feature 1407..1448 /note="MCS; multiple cloning site; unique BamHI, KpnI, SacI, and SnaBI cloning sites" gene 1440..2467 /gene="luxC" /label=luxC /note="Photorhabdus luminescens luxC cistron 5' end" regulatory 1440..1445 /gene="luxC" /regulatory_class="ribosome_binding_site" misc_recomb 1451..2467 /note="flanking region for double homologous recombination in Helicobacter pylori G27 vac::aphA-3/luxCDABE acceptor strain" CDS 1451..2467 /codon_start=1 /gene="luxC" /product="LuxC" /label=luxC /protein_id="ADZ38983.1" /translation="MTKKISFIINGQVEIFPESDDLVQSINFGDNSVYLPILNDSHVKN IIDCNGNNELRLHNIVNFLYTVGQRWKNEEYSRRRTYIRDLKKYMGYSEEMAKLEANWI SMILCSKGGLYDVVENELGSRHIMDEWLPQDESYVRAFPKGKSVHLLAGNVPLSGIMSI LRAILTKNQCIIKTSSTDPFTANALALSFIDVDPNHPITRSLSVIYWPHQGDTSLAKEI MRHADVIVAWGGPDAINWAVEHAPSYADVIKFGSKKSLCIIDNPVDLTSAATGAAHDVC FYDQRACFSAQNIYYMGNHYEEFKLALIEKLNLYAHILPNAKKDFDEKAAYSLVQKES" promoter complement(2476..2494) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2512..2528) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(2536..2552) /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(2560..2590) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(2605..2626) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(2914..3502) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3676..4533) /label=AmpR /note="beta-lactamase" promoter complement(4534..4638) /label=AmpR promoter primer_bind 5112..5128 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 5135..5153 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase"
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