LOCUS Exported 7937 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS HA-HIF1alpha P402A/P564A-pcDNA3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7937) 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 7937) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7937 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(46..65) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 237..616 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 617..820 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 771..791 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 865..884 /label=T7 /note="T7 promoter, forward primer" promoter 865..883 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 906..932 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" regulatory 945..954 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter complement(3492..3510) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(3493..3510) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(3530..3547) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3536..3760 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3806..4234 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3893..3912) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4103..4124 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4243..4263) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4248..4577 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4428..4563 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4490..4509 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4644..5438 /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(4698..4717) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5308..5327 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5612..5733 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5649..5668) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5703..5722 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5782..5798) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5782..5798) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5795..5817) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5806..5822 /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(5830..5860) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5875..5896 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6013..6030) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6184..6772) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6264..6283) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6943..7803) /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 7566..7585 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7804..7908) /gene="bla" /label=AmpR promoter ORIGIN 1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga 61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct 181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca 241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata 301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc 841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttacca 901 tggcctaccc ctacgacgtg cccgactacg cctccctcgg atccgccacc atggagggcg 961 ccggcggcgc gaacgacaag aaaaagataa gttctgaacg tcgaaaagaa aagtctcgag 1021 atgcagccag atctcggcga agtaaagaat ctgaagtttt ttatgagctt gctcatcagt 1081 tgccacttcc acataatgtg agttcgcatc ttgataaggc ctctgtgatg aggcttacca 1141 tcagctattt gcgtgtgagg aaacttctgg atgctggtga tttggatatt gaagatgaca 1201 tgaaagcaca gatgaattgc ttttatttga aagccttgga tggttttgtt atggttctca 1261 cagatgatgg tgacatgatt tacatttctg ataatgtgaa caaatacatg ggattaactc 1321 agtttgaact aactggacac agtgtgtttg attttactca tccatgtgac catgaggaaa 1381 tgagagaaat gcttacacac agaaatggcc ttgtgaaaaa gggtaaagaa caaaacacac 1441 agcgaagctt ttttctcaga atgaagtgta ccctaactag ccgaggaaga actatgaaca 1501 taaagtctgc aacatggaag gtattgcact gcacaggcca cattcacgta tatgatacca 1561 acagtaacca acctcagtgt gggtataaga aaccacctat gacctgcttg gtgctgattt 1621 gtgaacccat tcctcaccca tcaaatattg aaattccttt agatagcaag actttcctca 1681 gtcgacacag cctggatatg aaattttctt attgtgatga aagaattacc gaattgatgg 1741 gatatgagcc agaagaactt ttaggccgct caatttatga atattatcat gctttggact 1801 ctgatcatct gaccaaaact catcatgata tgtttactaa aggacaagtc accacaggac 1861 agtacaggat gcttgccaaa agaggtggat atgtctgggt tgaaactcaa gcaactgtca 1921 tatataacac caagaattct caaccacagt gcattgtatg tgtgaattac gttgtgagtg 1981 gtattattca gcacgacttg attttctccc ttcaacaaac agaatgtgtc cttaaaccgg 2041 ttgaatcttc agatatgaaa atgactcagc tattcaccaa agttgaatca gaagatacaa 2101 gtagcctctt tgacaaactt aagaaggaac ctgatgcttt aactttgctg gccgcagccg 2161 ctggagacac aatcatatct ttagattttg gcagcaacga cacagaaact gatgaccagc 2221 aacttgagga agtaccatta tataatgatg taatgctccc ctcacccaac gaaaaattac 2281 agaatataaa tttggcaatg tctccattac ccaccgctga aacgccaaag ccacttcgaa 2341 gtagtgctga ccctgcactc aatcaagaag ttgcattaaa attagaacca aatccagagt 2401 cactggaact ttcttttacc atgccccaga ttcaggatca gacacctagt ccttccgatg 2461 gaagcactag acaaagttca cctgagccta atagtcccag tgaatattgt ttttatgtgg 2521 atagtgatat ggtcaatgaa ttcaagttgg aattggtaga aaaacttttt gctgaagaca 2581 cagaagcaaa gaacccattt tctactcagg acacagattt agacttggag atgttagctg 2641 cttatatccc aatggatgat gacttccagt tacgttcctt cgatcagttg tcaccattag 2701 aaagcagttc cgcaagccct gaaagcgcaa gtcctcaaag cacagttaca gtattccagc 2761 agactcaaat acaagaacct actgctaatg ccaccactac cactgccacc actgatgaat 2821 taaaaacagt gacaaaagac cgtatggaag acattaaaat attgattgca tctccatctc 2881 ctacccacat acataaagaa actactagtg ccacatcatc accatataga gatactcaaa 2941 gtcggacagc ctcaccaaac agagcaggaa aaggagtcat agaacagaca gaaaaatctc 3001 atccaagaag ccctaacgtg ttatctgtcg ctttgagtca aagaactaca gttcctgagg 3061 aagaactaaa tccaaagata ctagctttgc agaatgctca gagaaagcga aaaatggaac 3121 atgatggttc actttttcaa gcagtaggaa ttggaacatt attacagcag ccagacgatc 3181 atgcagctac tacatcactt tcttggaaac gtgtaaaagg atgcaaatct agtgaacaga 3241 atggaatgga gcaaaagaca attattttaa taccctctga tttagcatgt agactgctgg 3301 ggcaatcaat ggatgaaagt ggattaccac agctgaccag ttatgattgt gaagttaatg 3361 ctcctataca aggcagcaga aacctactgc agggtgaaga attactcaga gctttggatc 3421 aagttaactg acaattctgc agatatccat cacactggcg gccgctcgag catgcatcta 3481 gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg 3541 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa 3601 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt 3661 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa 3721 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc 3781 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt 3841 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 3901 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 3961 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 4021 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 4081 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 4141 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa 4201 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag 4261 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 4321 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 4381 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 4441 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 4501 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 4561 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag 4621 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg 4681 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg 4741 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc 4801 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga 4861 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc 4921 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag 4981 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat 5041 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg 5101 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca 5161 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct 5221 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg 5281 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg 5341 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc 5401 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat 5461 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta 5521 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg 5581 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta 5641 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 5701 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag 5761 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 5821 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 5881 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 5941 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6001 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6061 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6121 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6181 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 6241 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 6301 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 6361 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 6421 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 6481 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 6541 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 6601 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 6661 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 6721 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 6781 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 6841 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 6901 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 6961 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7021 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7081 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7141 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7201 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 7261 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 7321 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 7381 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 7441 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 7501 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 7561 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 7621 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 7681 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 7741 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 7801 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 7861 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 7921 aaaagtgcca cctgacg //