Basic Vector Information
pUCTCAPR2-GW 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 ℃.
pUCTCAPR2-GW vector Sequence
LOCUS 40924_45398 8382 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector pUCTCAPR2-GW, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8382) AUTHORS Kvitko BH, McMillan IA, Schweizer HP. TITLE An Improved Method for oriT-Directed Cloning and Functionalization of Large Bacterial Genomic Regions JOURNAL Appl. Environ. Microbiol. 79 (16), 4869-4878 (2013) PUBMED 23747708 REFERENCE 2 (bases 1 to 8382) AUTHORS Kvitko BH, McMillan IA, Schweizer HP. TITLE Direct Submission JOURNAL Submitted (03-SEP-2012) Microbiology Immunology and Pathology, Colorado State University, IDRC at Foothills Campus, Campus Delivery 0922, Fort Collins, CO 80523-0922, USA REFERENCE 3 (bases 1 to 8382) TITLE Direct Submission REFERENCE 4 (bases 1 to 8382) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl. Environ. Microbiol."; date: "2013"; volume: "79"; issue: "16"; pages: "4869-4878" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (03-SEP-2012) Microbiology Immunology and Pathology, Colorado State University, IDRC at Foothills Campus, Campus Delivery 0922, Fort Collins, CO 80523-0922, USA" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..8382 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 1..372 /codon_start=1 /label=BleoR /note="antibiotic-binding protein" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" misc_recomb 486..533 /label=FRT /note="FRT" protein_bind complement(486..533) /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." terminator complement(586..672) /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator complement(775..869) /label=lambda t0 terminator /note="transcription terminator from phage lambda" primer_bind complement(897..913) /label=KS primer /note="common sequencing primer, one of multiple similar variants" repeat_region 918..1116 /label=Tn7R /note="Tn7R" misc_feature 1264..1281 /label=I-SceI restriction site /note="I-SceI restriction site" primer_bind complement(1291..1307) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" terminator 1657..1743 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 1835..1862 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" oriT 2309..2418 /label=oriT /note="incP origin of transfer" terminator complement(3117..3160) /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 3292..3319 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 3338..3429 /label=AmpR promoter CDS complement(3895..4425) /codon_start=1 /label=GmR /note="gentamycin acetyltransferase" /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" promoter complement(4614..4642) /label=Pc promoter /note="class 1 integron promoter" rep_origin 5365..5953 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 6241..6262 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 6277..6307 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6315..6331 /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)." primer_bind 6339..6355 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" misc_feature 6374..6391 /label=I-SceI restriction site /note="I-SceI restriction site" protein_bind 6414..6538 /label=attR1 /note="recombination site for the Gateway(R) LR reaction" promoter 6563..6593 /label=lac UV5 promoter /note="E. coli lac promoter with an 'up' mutation" CDS 6647..7303 /codon_start=1 /label=CmR /note="chloramphenicol acetyltransferase" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" CDS 7648..7950 /codon_start=1 /label=ccdB /note="CcdB, a bacterial toxin that poisons DNA gyrase" /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" protein_bind complement(7994..8118) /label=attR2 /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(8180..8227) /label=FRT /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." promoter 8317..8364 /label=EM7 promoter /note="synthetic bacterial promoter"
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