pcDNA-NBid-mMaple-CBid vector (V015772)

LOCUS       62056_5385        6709 bp DNA     circular SYN 22-JUN-2021
DEFINITION  Cloning vector pcDNA-NBid-mMaple-CBid, complete sequence.
ACCESSION   MW307777
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6709)
  AUTHORS   Lu X.
  TITLE     Improved Photocleavable Proteins with Faster and More Efficient 
            Dissociation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6709)
  AUTHORS   Lu X.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-NOV-2020) Department of Chemistry, University of 
            Alberta, ccis4-140,11227 Saskatchewan Drive, Edmonton, AB T6G2G2, 
            Canada
REFERENCE   3  (bases 1 to 6709)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-NOV-2020) Department of Chemistry, University of Alberta, 
            ccis4-140,11227 Saskatchewan Drive, Edmonton, AB T6G2G2, Canada"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: synthetic gene
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1081..1788
                     /codon_start=1
                     /label=mMaple
                     /note="monomeric photoconvertible green-to-red fluorescent 
                     protein derived from mClavGR2 (McEvoy et al., 2012)"
                     /translation="VSKGEETIMSVIKPDMKIKLRMEGNVNGHAFVIEGEGSGKPFEGI
                     QTIDLEVKEGAPLPFAYDILTTAFHYGNRVFTKYPEDIPDYFKQSFPEGYSWERSMTYE
                     DGGICIATNDITMEEDSFINKIHFKGTNFPPNGPVMQKRTVGWEVSTEKMYVRDGVLKG
                     DVKMKLLLKGGSHYRCDFRTTYKVKQKAVKLPDYHFVDHRIEILSHDKDYNKVKLYEHA
                     VARNSTDSMDELYK"
     polyA_signal    2309..2533
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2579..3007
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3021..3350
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3417..4208
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4385..4518
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4555..4571)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4579..4595)
                     /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(4603..4633)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4648..4669)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4957..5542)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5716..6573)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6574..6678)
                     /label=AmpR promoter