Basic Vector Information
- Vector Name:
- PCY
- Antibiotic Resistance:
- Kanamycin
- Length:
- 10347 bp
- Type:
- Shuttle vector
- Replication origin:
- ori
- Source/Author:
- Cui T, Song Z.
- Promoter:
- TRP1
PCY 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 ℃.
PCY vector Sequence
LOCUS 40924_14065 10347 bp DNA circular SYN 17-DEC-2018 DEFINITION Shuttle vector PCY, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10347) AUTHORS Cui T, Song Z. TITLE Direct Submission JOURNAL Submitted (05-DEC-2017) Citrus Research Institute, Southwest University, Xie ma, Beibei, Chongqing 400712, China REFERENCE 2 (bases 1 to 10347) TITLE Direct Submission REFERENCE 3 (bases 1 to 10347) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted (05-DEC-2017) Citrus Research Institute, Southwest University, Xie ma, Beibei, Chongqing 400712, China" COMMENT SGRef: number: 2; type: "Journal Article" COMMENT ##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..10347 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 95..767 /label=CaMV 35S promoter (enhanced) /note="cauliflower mosaic virus 35S promoter with a duplicated enhancer region" CDS 771..1484 /codon_start=1 /label=mgfp5 /note="GFP with folding enhancement mutations" /translation="MSKGEELFTGVVPILVELDGDVNGYKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNFHNVYIMADKQKNGIKA NFKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" terminator 1589..1841 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" misc_feature 1863..1887 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 3187..3813 /codon_start=1 /label=pVS1 StaA /note="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" CDS 4250..5314 /codon_start=1 /label=pVS1 RepA /note="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV MRYRNLIEGEASAGS" rep_origin 5383..5577 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" misc_feature 5921..6061 /label=bom /note="basis of mobility region from pBR322" rep_origin complement(6247..6835) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6925..7716) /codon_start=1 /label=KanR /note="aminoglycoside phosphotransferase" /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF" CDS complement(8058..8729) /codon_start=1 /label=TRP1 /note="phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis" /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQLKEDVLALVNDYGIDIVQLHGDES WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA KK" promoter complement(8730..9011) /label=TRP1 promoter misc_feature complement(9269..9772) /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" promoter 9809..9913 /label=AmpR promoter misc_feature 10258..10282 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA"
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