Basic Vector Information
- Vector Name:
- CMV+-mScarlet-I-with-onboard-mCitrine-cassette
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9294 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Yang J, Lee J, Land MA, Lai S
- Promoter:
- EF-1α
CMV+-mScarlet-I-with-onboard-mCitrine-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+-mScarlet-I-with-onboard-mCitrine-cassette vector Sequence
LOCUS 62056_536 9294 bp DNA circular SYN 18-OCT-2021
DEFINITION Cloning vector CMV+ mScarlet-I with onboard mCitrine cassette,
complete sequence.
ACCESSION MW987532
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9294)
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JOURNAL Unpublished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 9294)
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 9294)
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"Unpublished"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##
On Oct 18, 2021 this sequence version replaced MW987532.1.
FEATURES Location/Qualifiers
source 1..9294
/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..1873
/codon_start=1
/label=mScarlet-I
/note="bright monomeric red fluorescent protein evolved
from a synthetic template (Bindels et al., 2016)"
/translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
STGGMDELYK"
polyA_signal 2317..2541
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
protein_bind 2825..2872
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS 2880..3899
/codon_start=1
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
/translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
"
polyA_signal 4032..4165
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4202..4218)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4226..4242)
/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(4250..4280)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4295..4316)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4431..5612
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 5634..6350
/codon_start=1
/label=mEYFP
/note="enhanced YFP with monomerizing A206K mutation
(Zacharias et al., 2002)"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
promoter complement(6372..6390)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
rep_origin complement(7086..7674)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7848..8705)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(8706..8810)
/label=AmpR promoter
enhancer 9076..9294
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
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