Basic Vector Information
- Vector Name:
- EK0294 SFFV-mCherry-SG(s70587-W)SG-EGFP (FLP-IN)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6590 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells
- Selection Marker:
- Hyg
- Promoter:
- SFFV
- 5' Primer:
- BGH
- Growth Temperature:
- 37℃
EK0294 SFFV-mCherry-SG(s70587-W)SG-EGFP (FLP-IN) 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 ℃.
EK0294 SFFV-mCherry-SG(s70587-W)SG-EGFP (FLP-IN) vector Sequence
LOCUS 62056_681 6590 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6590) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..6590 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 90..497 /label=SFFV promoter /note="spleen focus-forming virus long terminal repeat (LTR) promoter" CDS 511..1218 /codon_start=1 /label=mCherry /note="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" CDS 1522..2235 /codon_start=1 /label=EGFP /note="enhanced GFP" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" CDS 2257..2286 /codon_start=1 /label=Myc /note="Myc (human c-Myc proto-oncogene) epitope tag" /translation="EQKLISEEDL" polyA_signal 2348..2572 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" protein_bind 2895..2942 /label=FRT /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS 2950..3969 /codon_start=1 /label=HygR /note="aminoglycoside phosphotransferase from E. coli" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 4102..4235 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4272..4288) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(4296..4312) /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(4320..4350) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(4365..4386) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4674..5262) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(5436..6293) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(6294..6398) /label=AmpR promoter promoter 6590 /label=SFFV /note="Spleen focus-forming virus promoter"
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