Basic Vector Information
pUCP30T-mOrange 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 ℃.
pUCP30T-mOrange vector Sequence
LOCUS 40924_45383 5050 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector pUCP30T-mOrange, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5050) 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 5050) 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 5050) TITLE Direct Submission REFERENCE 4 (bases 1 to 5050) 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..5050 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin 72..660 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" oriT complement(732..840) /direction=LEFT /label=oriT /note="incP origin of transfer" protein_bind 1224..1245 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 1260..1290 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1298..1314 /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 1322..1338 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" CDS 1378..1395 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" CDS 1405..2112 /codon_start=1 /label=LSSmOrange /note="monomeric orange fluorescent protein with a large Stokes shift (Shcherbakova et al., 2012)" /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG FQTVKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGALK GEDKLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(2155..2171) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS complement(2335..3165) /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 3179..3530 /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)" promoter 3571..3599 /label=Pc promoter /note="class 1 integron promoter" CDS 3788..4318 /codon_start=1 /label=GmR /note="gentamycin acetyltransferase" /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT"
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