pUbC-mNeonGreen4-tDeg vector (V016357)

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LOCUS       62056_21820        8122 bp DNA     circular SYN 10-SEP-2019
DEFINITION  Cloning vector pUbC-mNeonGreen4-tDeg, complete sequence.
ACCESSION   MN052907
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8122)
  AUTHORS   Wu J, Zaccara S, Khuperkar D, Kim H, Tanenbaum ME, Jaffrey SR.
  TITLE     Live imaging of mRNA using RNA-stabilized fluorogenic proteins
  JOURNAL   Nat. Methods 16 (9), 862-865 (2019)
  PUBMED    31471614
REFERENCE   2  (bases 1 to 8122)
  AUTHORS   Wu J, Zaccara S, Khuperkar D, Kim H, Tanenbaum ME, Jaffrey SR.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JUN-2019) Pharmacology, Weill Cornell Medicine, 1300 
            York Avenue, New York, NY 10022, United States
REFERENCE   3  (bases 1 to 8122)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Methods"; date: "2019"; volume: "16"; issue: "9"; pages: "862-865"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-JUN-2019) Pharmacology, Weill Cornell Medicine, 1300 York 
            Avenue, New York, NY 10022, United States"
FEATURES             Location/Qualifiers
     source          1..8122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        235..633
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     CDS             646..1353
                     /codon_start=1
                     /label=mNeonGreen
                     /note="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     polyA_signal    3719..3946
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3992..4420
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4434..4763
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4830..5621
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5798..5931
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5968..5984)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5992..6008)
                     /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(6016..6046)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6061..6082)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6370..6955)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7129..7986)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7987..8091)
                     /label=AmpR promoter