LOCUS       sequence-275563-        7931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HIF1a P402A/P564A/N803A mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7931)
  AUTHORS   Yan Q, Bartz S, Mao M, Li L, Kaelin WG
  TITLE     The hypoxia-inducible factor 2alpha N-terminal and C-terminal 
            transactivation domains cooperate to promote renal tumorigenesis in 
            vivo.
  JOURNAL   Mol Cell Biol. 2007 Mar . 27(6):2092-102.
  PUBMED    17220275
REFERENCE   2  (bases 1 to 7931)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell 
            Biol. 2007 Mar . 27(6):2092-102."
COMMENT     Sequence Label: HA-HIF1-alpha P402A/P564A/N803A-pcDNA3
FEATURES             Location/Qualifiers
     source          1..7931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        151..530
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        531..734
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     685..705
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     779..798
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        779..797
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             820..846
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      853..862
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3400..3418)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3401..3418)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3438..3455)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3444..3668
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3714..4142
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3801..3820)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4011..4032
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4151..4171)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4156..4485
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4336..4471
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4398..4417
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4552..5346
                     /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"
     primer_bind     complement(4606..4625)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5216..5235
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5520..5641
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5557..5576)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5611..5630
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5690..5706)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5690..5706)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5703..5725)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5714..5730
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(join(5738..5744,5745..5762,5763..5768))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5783..5804
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5921..5938)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6092..6680)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6172..6191)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6851..7642,7643..7711))
                     /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"
     primer_bind     7474..7493
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7712..7816)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7891..7910)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
       61 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
      121 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
      181 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      241 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      301 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      361 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      421 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      481 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      541 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      601 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      661 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      721 atataagcag agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta
      781 atacgactca ctatagggag acccaagctt gccaccatgt atccatatga tgttccagat
      841 tatgctggat ccgccaccat ggagggcgcc ggcggcgcga acgacaagaa aaagataagt
      901 tctgaacgtc gaaaagaaaa gtctcgagat gcagccagat ctcggcgaag taaagaatct
      961 gaagtttttt atgagcttgc tcatcagttg ccacttccac ataatgtgag ttcgcatctt
     1021 gataaggcct ctgtgatgag gcttaccatc agctatttgc gtgtgaggaa acttctggat
     1081 gctggtgatt tggatattga agatgacatg aaagcacaga tgaattgctt ttatttgaaa
     1141 gccttggatg gttttgttat ggttctcaca gatgatggtg acatgattta catttctgat
     1201 aatgtgaaca aatacatggg attaactcag tttgaactaa ctggacacag tgtgtttgat
     1261 tttactcatc catgtgacca tgaggaaatg agagaaatgc ttacacacag aaatggcctt
     1321 gtgaaaaagg gtaaagaaca aaacacacag cgaagctttt ttctcagaat gaagtgtacc
     1381 ctaactagcc gaggaagaac tatgaacata aagtctgcaa catggaaggt attgcactgc
     1441 acaggccaca ttcacgtata tgataccaac agtaaccaac ctcagtgtgg gtataagaaa
     1501 ccacctatga cctgcttggt gctgatttgt gaacccattc ctcacccatc aaatattgaa
     1561 attcctttag atagcaagac tttcctcagt cgacacagcc tggatatgaa attttcttat
     1621 tgtgatgaaa gaattaccga attgatggga tatgagccag aagaactttt aggccgctca
     1681 atttatgaat attatcatgc tttggactct gatcatctga ccaaaactca tcatgatatg
     1741 tttactaaag gacaagtcac cacaggacag tacaggatgc ttgccaaaag aggtggatat
     1801 gtctgggttg aaactcaagc aactgtcata tataacacca agaattctca accacagtgc
     1861 attgtatgtg tgaattacgt tgtgagtggt attattcagc acgacttgat tttctccctt
     1921 caacaaacag aatgtgtcct taaaccggtt gaatcttcag atatgaaaat gactcagcta
     1981 ttcaccaaag ttgaatcaga agatacaagt agcctctttg acaaacttaa gaaggaacct
     2041 gatgctttaa ctttgctggc cgcagccgct ggagacacaa tcatatcttt agattttggc
     2101 agcaacgaca cagaaactga tgaccagcaa cttgaggaag taccattata taatgatgta
     2161 atgctcccct cacccaacga aaaattacag aatataaatt tggcaatgtc tccattaccc
     2221 accgctgaaa cgccaaagcc acttcgaagt agtgctgacc ctgcactcaa tcaagaagtt
     2281 gcattaaaat tagaaccaaa tccagagtca ctggaacttt cttttaccat gccccagatt
     2341 caggatcaga cacctagtcc ttccgatgga agcactagac aaagttcacc tgagcctaat
     2401 agtcccagtg aatattgttt ttatgtggat agtgatatgg tcaatgaatt caagttggaa
     2461 ttggtagaaa aactttttgc tgaagacaca gaagcaaaga acccattttc tactcaggac
     2521 acagatttag acttggagat gttagctgct tatatcccaa tggatgatga cttccagtta
     2581 cgttccttcg atcagttgtc accattagaa agcagttccg caagccctga aagcgcaagt
     2641 cctcaaagca cagttacagt attccagcag actcaaatac aagaacctac tgctaatgcc
     2701 accactacca ctgccaccac tgatgaatta aaaacagtga caaaagaccg tatggaagac
     2761 attaaaatat tgattgcatc tccatctcct acccacatac ataaagaaac tactagtgcc
     2821 acatcatcac catatagaga tactcaaagt cggacagcct caccaaacag agcaggaaaa
     2881 ggagtcatag aacagacaga aaaatctcat ccaagaagcc ctaacgtgtt atctgtcgct
     2941 ttgagtcaaa gaactacagt tcctgaggaa gaactaaatc caaagatact agctttgcag
     3001 aatgctcaga gaaagcgaaa aatggaacat gatggttcac tttttcaagc agtaggaatt
     3061 ggaacattat tacagcagcc agacgatcat gcagctacta catcactttc ttggaaacgt
     3121 gtaaaaggat gcaaatctag tgaacagaat ggaatggagc aaaagacaat tattttaata
     3181 ccctctgatt tagcatgtag actgctgggg caatcaatgg atgaaagtgg attaccacag
     3241 ctgaccagtt atgattgtga agttgctgct cctatacaag gcagcagaaa cctactgcag
     3301 ggtgaagaat tactcagagc tttggatcaa gttaactgac aattctgcag atatccatca
     3361 cactggcggc cgctcgagca tgcatctaga gggccctatt ctatagtgtc acctaaatgc
     3421 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3481 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3541 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3601 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3661 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3721 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3781 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3841 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3901 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3961 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4021 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4081 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4141 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4201 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4261 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4321 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4381 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4441 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4501 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4561 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4621 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4681 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4741 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4801 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4861 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4921 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4981 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5041 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5101 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5161 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5221 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5281 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5341 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5401 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5461 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5521 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5581 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5641 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5701 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5761 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5821 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5881 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5941 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6001 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6061 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6121 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6181 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6241 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6301 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6361 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6421 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6481 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6541 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6601 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6661 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6721 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6781 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6841 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6901 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6961 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7021 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7081 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7141 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7201 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7261 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7321 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7381 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7441 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7501 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7561 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7621 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7681 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7741 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7801 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     7861 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     7921 ccagtatctg c
//
