Basic Vector Information
2C::tdTomato Reporter 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 ℃.
2C::tdTomato Reporter vector Sequence
LOCUS 40924_31 7132 bp DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7132) AUTHORS Macfarlan TS, Gifford WD, Driscoll S, Lettieri K, Rowe HM, Bonanomi D, Firth A, Singer O, Trono D, Pfaff SL TITLE Embryonic stem cell potency fluctuates with endogenous retrovirus activity. JOURNAL Nature. 2012 Jul 5;487(7405):57-63. PUBMED 22722858 REFERENCE 2 (bases 1 to 7132) TITLE Direct Submission REFERENCE 3 (bases 1 to 7132) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature."; date: "2012-07-5"; volume: "487(7405)"; pages: "57-63" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..7132 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind complement(75..94) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" CDS 1067..2494 /codon_start=1 /label=tdTomato /note="tandem dimeric (pseudo-monomeric) derivative of DsRed (Shaner et al., 2004)" /translation="MVSKGEDVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE RSEGRHHLFLYGMDELYK" polyA_signal 2591..2815 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2861..3289 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3303..3632 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 3681..4703 /codon_start=1 /label=HygR /note="aminoglycoside phosphotransferase from E. coli" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" polyA_signal 4836..4969 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5006..5022) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(5030..5046) /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(5054..5084) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(5099..5120) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5237..5254) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5408..5996) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6170..7027) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(7028..7132) /label=AmpR promoter
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