Basic Vector Information
- Vector Name:
- CMV+-with-onboard-mCherry-cassette
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9314 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Yang J, Lee J, Land MA, Lai S
- Promoter:
- EF-1α
CMV+-with-onboard-mCherry-cassette 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 ℃.
CMV+-with-onboard-mCherry-cassette vector Sequence
LOCUS 62056_541 9314 bp DNA circular SYN 14-MAY-2021 DEFINITION Cloning vector CMV+ with onboard mCherry cassette, complete sequence. ACCESSION MW987530 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9314) AUTHORS Yang J, Lee J, Land MA, Lai S, Igoshin O, St-Pierre F. TITLE A synthetic circuit for buffering gene dosage variation between individual mammalian cells JOURNAL Unpublished REFERENCE 2 (bases 1 to 9314) AUTHORS Yang J, Lee J, Land MA, Lai S, Igoshin O, St-Pierre F. TITLE Direct Submission JOURNAL Submitted (19-APR-2021) Neuroscience, Baylor College of Medicine, One Baylor Plaza, Suite S636, Houston, Texas 77030, United States REFERENCE 3 (bases 1 to 9314) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (19-APR-2021) Neuroscience, Baylor College of Medicine, One Baylor Plaza, Suite S636, Houston, Texas 77030, United States" COMMENT ##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..9314 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 163..360 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" protein_bind 361..379 /label=tet operator /note="bacterial operator O2 for the tetR and tetA genes" intron 588..1160 /label=beta-globin intron /note="intron from rabbit beta-globin gene" CDS 1178..1894 /codon_start=1 /label=EGFP /note="enhanced GFP" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" polyA_signal 2338..2562 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" protein_bind 2846..2893 /label=FRT /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS 2901..3920 /codon_start=1 /label=HygR /note="aminoglycoside phosphotransferase from E. coli" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 4053..4186 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4223..4239) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(4247..4263) /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(4271..4301) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(4316..4337) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 4584..5762 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" CDS 5807..6514 /codon_start=1 /label=mCherry /note="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" polyA_signal 6674..6729 /label=beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" rep_origin complement(7106..7694) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(7868..8725) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(8726..8830) /label=AmpR promoter enhancer 9096..9314 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer"
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