Basic Vector Information
pUCP20T-mPlum 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 ℃.
pUCP20T-mPlum vector Sequence
LOCUS 40924_45343 4854 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector pUCP20T-mPlum, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4854) AUTHORS Barbier M, Damron FH. TITLE Rainbow Vectors for Broad-Range Bacterial Fluorescence Labeling JOURNAL PLoS ONE 11 (3), E0146827 (2016) PUBMED 26937640 REFERENCE 2 (bases 1 to 4854) AUTHORS Barbier M, Damron FH. TITLE Direct Submission JOURNAL Submitted (06-OCT-2015) Microbiology, Immunology and Cell Biology, West Virginia University, School of Medicine, One Medical Center Drive, Morgantown, WV 26506, USA REFERENCE 3 (bases 1 to 4854) TITLE Direct Submission REFERENCE 4 (bases 1 to 4854) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; date: "2016"; volume: "11"; issue: "3"; pages: "E0146827" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (06-OCT-2015) Microbiology, Immunology and Cell Biology, West Virginia University, School of Medicine, One Medical Center Drive, Morgantown, WV 26506, USA" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..4854 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 96..200 /label=AmpR promoter CDS 201..1058 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 1232..1820 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" oriT complement(1892..2000) /direction=LEFT /label=oriT /note="incP origin of transfer" protein_bind 2384..2405 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 2420..2450 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2458..2474 /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 2482..2498 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" CDS 2528..3205 /codon_start=1 /label=mPlum /note="far-red monomeric derivative of DsRed fluorescent protein (Wang et al., 2004)" /translation="MVSKGEEVIKEFMRFKEHMEGSVNGHEFEIEGEGEGRPYEGTQTA RLKVTKGGPLPFAWDILSPQIMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGG VVTVTQDSSLQDGEFIYKVKVRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEMK MRLRLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRH STGA" primer_bind complement(3248..3264) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS complement(3428..4258) /codon_start=1 /label=pRO1600 Rep /note="replication protein for the broad-host-range plasmid pRO1600 from Pseudomonas aeruginosa" /translation="VASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA" rep_origin 4272..4623 /label=pRO1600 oriV /note="broad-host-range origin of replication from Pseudomonas aeruginosa plasmid pRO1600; requires the pRO1600 Rep protein for replication (West et al., 1994)"
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