LOCUS       Exported                7858 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCMV-HIF-1a-FLAG, complete sequence.
ACCESSION   GQ340758
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7858)
  AUTHORS   Cook KM, Hilton ST, Mecinovic J, Motherwell WB, Figg WD, Schofield 
            CJ.
  TITLE     Epidithiodiketopiperazines block the interaction between 
            hypoxia-inducible factor-1alpha (HIF-1alpha) and p300 by a zinc 
            ejection mechanism
  JOURNAL   J. Biol. Chem. 284 (39), 26831-26838 (2009)
  PUBMED    19589782
REFERENCE   2  (bases 1 to 7858)
  AUTHORS   Figg WD, Chau CH, Cook KM.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-JUL-2009) NCI, NIH, 10 Center Drive, Bldg 10, Rm 5A01,
            Bethesda, MD 20892, USA
REFERENCE   3  (bases 1 to 7858)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7858
                     /organism="Cloning vector pCMV-HIF-1a-FLAG"
                     /mol_type="other DNA"
                     /db_xref="taxon:663330"
     regulatory      1..523
                     /regulatory_class="promoter"
                     /note="CMV"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     634..650
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      673..691
                     /regulatory_class="promoter"
                     /note="SP6"
     promoter        673..691
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     protein_bind    716..740
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    743..3289
                     /gene="HIF1A"
                     /label=HIF-1a-flag
                     /note="HIF-1a-flag"
     CDS             743..3289
                     /codon_start=1
                     /gene="HIF1A"
                     /product="hypoxia inducible factor 1 subunit alpha"
                     /label=HIF1A
                     /note="transcription factor; HIF-1a; derived from Homo 
                     sapiens"
                     /protein_id="ACT98611.1"
                     /translation="MEGAGGANDKKKISSERRKEKSRDAARSRRSKESEVFYELAHQLP
                     LPHNVSSHLDKASVMRLTISYLRVRKLLDAGDLDIEDDMKAQMNCFYLKALDGFVMVLT
                     DDGDMIYISDNVNKYMGLTQFELTGHSVFDFTHPCDHEEMREMLTHRNGLVKKGKEQNT
                     QRSFFLRMKCTLTSRGRTMNIKSATWKVLHCTGHIHVYDTNSNQPQCGYKKPPMTCLVL
                     ICEPIPHPSNIEIPLDSKTFLSRHSLDMKFSYCDERITELMGYEPEELLGRSIYEYYHA
                     LDSDHLTKTHHDMFTKGQVTTGQYRMLAKRGGYVWVETQATVIYNTKNSQPQCIVCVNY
                     VVSGIIQHDLIFSLQQTECVLKPVESSDMKMTQLFTKVESEDTSSLFDKLKKEPDALTL
                     LAPAAGDTIISLDFGSNDTETDDQQLEEVPLYNDVMLPSPNEKLQNINLAMSPLPTAET
                     PKPLRSSADPALNQEVALKLEPNPESLELSFTMPQIQDQTPSPSDGSTRQSSPEPNSPS
                     EYCFYVDSDMVNEFKLELVEKLFAEDTEAKNPFSTQDTDLDLEMLAPYIPMDDDFQLRS
                     FDQLSPLESSSASPESASPQSTVTVFQQTQIQEPTANATTTTATTDELKTVTKDRMEDI
                     KILIASPSPTHIHKETTSATSSPYRDTQSRTASPNRAGKGVIEQTEKSHPRSPNVLSVA
                     LSQRTTVPEEELNPKILALQNAQRKRKMEHDGSLFQAVGIGTLLQQPDDHAATTSLSWK
                     RVKGCKSSEQNGMEQKTIILIPSDLACRLLGQSMDESGLPQLTSYDCEVNAPIQGSRNL
                     LQGEELLRALDQVNPTFLYKVVRSRMASDYKDDDDK"
     gene            743..3289
                     /gene="HIF1A"
                     /label=HIF1A
     protein_bind    complement(3219..3243)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3263..3286
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3616..3840
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3886..4314
                     /label=f1
                     /note="f1"
     promoter        4328..4657
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4341..4650
                     /label=SV40
                     /note="SV40"
     rep_origin      4508..4643
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4724..5518
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    4725..5518
                     /label=Neo(R)
                     /note="Neo(R)"
     polyA_signal    5582..5630
                     /note="synthetic polyadenylation signal"
     promoter        5936..6040
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6041..6921
                     /label=bla(R)
                     /note="bla(R)"
     CDS             6041..6901
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7050..7689
     rep_origin      7072..7660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"