LOCUS       Exported                8043 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    HA-HIF2alpha-P405A/P531A-pcDNA3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8043)
  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 8043)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8043
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     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"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             904..930
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3595..3613)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3596..3613)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3633..3650)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3639..3863
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3909..4337
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3996..4015)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4206..4227
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4346..4366)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4351..4681
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4532..4667
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4594..4613
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4748..5542
                     /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(4802..4821)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5412..5431
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5716..5837
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5753..5772)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5807..5826
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5886..5902)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5886..5902)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5899..5921)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5910..5926
                     /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(5934..5964)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5979..6000
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6117..6134)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6288..6876)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6368..6387)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7047..7907)
                     /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     7670..7689
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7908..8012)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
      901 atgtatccat atgatgttcc agattatgct ggatccgcca ccatgacagc tgacaaggag
      961 aagaaaagga gtagctcgga gaggaggaag gagaagtccc gggatgctgc gcggtgccgg
     1021 cggagcaagg agacggaggt gttctatgag ctggcccatg agctgcctct gccccacagt
     1081 gtgagctccc atctggacaa ggcctccatc atgcgactgg caatcagctt cctgcgaaca
     1141 cacaagctcc tctcctcagt ttgctctgaa aacgagtccg aagccgaagc tgaccagcag
     1201 atggacaact tgtacctgaa agccttggag ggtttcattg ccgtggtgac ccaagatggc
     1261 gacatgatct ttctgtcaga aaacatcagc aagttcatgg gacttacaca ggtggagcta
     1321 acaggacata gtatctttga cttcactcat ccctgcgacc atgaggagat tcgtgagaac
     1381 ctgagtctca aaaatggctc tggttttggg aaaaaaagca aagacatgtc cacagagcgg
     1441 gacttcttca tgaggatgaa gtgcacggtc accaacagag gccgtactgt caacctcaag
     1501 tcagccacct ggaaggtctt gcactgcacg ggccaggtga aagtctacaa caactgccct
     1561 cctcacaata gtctgtgtgg ctacaaggag cccctgctgt cctgcctcat catcatgtgt
     1621 gaaccaatcc agcacccatc ccacatggac atccccctgg atagcaagac cttcctgagc
     1681 cgccacagca tggacatgaa gttcacctac tgtgatgaca gaatcacaga actgattggt
     1741 taccaccctg aggagctgct tggccgctca gcctatgaat tctaccatgc gctagactcc
     1801 gagaacatga ccaagagtca ccagaacttg tgcaccaagg gtcaggtagt aagtggccag
     1861 taccggatgc tcgcaaagca tgggggctac gtgtggctgg agacccaggg gacggtcatc
     1921 tacaaccctc gcaacctgca gccccagtgc atcatgtgtg tcaactacgt cctgagtgag
     1981 attgagaaga atgacgtggt gttctccatg gaccagactg aatccctgtt caagccccac
     2041 ctgatggcca tgaacagcat ctttgatagc agtggcaagg gggctgtgtc tgagaagagt
     2101 aacttcctat tcaccaagct aaaggaggag cccgaggagc tggcccagct ggctgctacc
     2161 ccaggagacg ccatcatctc tctggatttc gggaatcaga acttcgagga gtcctcagcc
     2221 tatggcaagg ccatcctgcc cccgagccag ccatgggcca cggagttgag gagccacagc
     2281 acccagagcg aggctgggag cctgcctgcc ttcaccgtgc cccaggcagc tgccccgggc
     2341 agcaccaccc ccagtgccac cagcagcagc agcagctgct ccacgcccaa tagccctgaa
     2401 gactattaca catctttgga taacgacctg aagattgaag tgattgagaa gctcttcgcc
     2461 atggacacag aggccaagga ccaatgcagt acccagacgg atttcaatga gctggacttg
     2521 gagacactgg cagcttatat ccccatggac ggggaagact tccagctaag ccccatctgc
     2581 cccgaggagc ggctcttggc ggagaaccca cagtccaccc cccagcactg cttcagtgcc
     2641 atgacaaaca tcttccagcc actggcccct gtagccccgc acagtccctt cctcctggac
     2701 aagtttcagc agcagctgga gagcaagaag acagagcccg agcaccggcc catgtcctcc
     2761 atcttctttg atgccggaag caaagcatcc ctgccaccgt gctgtggcca ggccagcacc
     2821 cctctctctt ccatgggggg cagatccaat acccagtggc ccccagatcc accattacat
     2881 tttgggccca caaagtgggc cgtcggggat cagcgcacag agttcttggg agcagcgccg
     2941 ttggggcccc ctgtctctcc accccatgtc tccaccttca agacaaggtc tgcaaagggt
     3001 tttggggctc gaggcccaga cgtgctgagt ccggccatgg tagccctctc caacaagctg
     3061 aagctgaagc gacagctgga gtatgaagag caagccttcc aggacctgag cgggggggac
     3121 ccacctggtg gcagcacctc acatttgatg tggaaacgga tgaagaacct caggggtggg
     3181 agctgccctt tgatgccgga caagccactg agcgcaaatg tacccaatga taagttcacc
     3241 caaaacccca tgaggggcct gggccatccc ctgagacatc tgccgctgcc acagcctcca
     3301 tctgccatca gtcccgggga gaacagcaag agcaggttcc ccccacagtg ctacgccacc
     3361 cagtaccagg actacagcct gtcgtcagcc cacaaggtgt caggcatggc aagccggctg
     3421 ctcgggccct catttgagtc ctacctgctg cccgaactga ccagatatga ctgtgaggtg
     3481 aacgtgcccg tgctgggaag ctccacgctc ctgcaaggag gggacctcct cagagccctg
     3541 gaccaggcca cctgacaatt gcggccgctc gagcatgcat ctagagggcc ctattctata
     3601 gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     3661 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3721 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3781 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3841 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3901 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3961 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4021 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     4081 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4141 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4201 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4261 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     4321 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4381 ggctccccag gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4441 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4501 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4561 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4621 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4681 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     4741 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4801 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4861 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4921 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4981 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     5041 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     5101 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5161 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5221 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     5281 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5341 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5401 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5461 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5521 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5581 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5641 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5701 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5761 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5821 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5881 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5941 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6001 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6061 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6121 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6181 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6241 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6301 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6361 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6421 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6481 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6541 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6601 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6661 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6721 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6781 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6841 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6901 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6961 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7021 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7081 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7141 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7201 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7261 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7321 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7381 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7441 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7501 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7561 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7621 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7681 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7741 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7801 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7861 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7921 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7981 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8041 gtc
//
