CMV+-mScarlet-I-with-onboard-mCitrine-cassette vector (V015778)

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  JOURNAL   Unpublished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            "Unpublished"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"