LOCUS Exported 8069 bp ds-DNA circular SYN 16-AUG-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS HA-HIF2alpha-pcDNA3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8069) AUTHORS Kondo K, Klco J, Nakamura E, Lechpammer M, Kaelin WG TITLE Inhibition of HIF is necessary for tumor suppression by the von Hippel-Lindau protein. JOURNAL Cancer Cell. 2002 Apr . 1(3):237-46. PUBMED 12086860 REFERENCE 2 (bases 1 to 8069) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 16, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8069 /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(3622..3640) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(3623..3640) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(3660..3677) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3666..3890 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3936..4364 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4023..4042) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4233..4254 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4373..4393) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4378..4707 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4558..4693 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4620..4639 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4774..5568 /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(4828..4847) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5438..5457 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5742..5863 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5779..5798) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5833..5852 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5912..5928) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5912..5928) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5925..5947) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5936..5952 /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(5960..5990) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6005..6026 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6143..6160) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6314..6902) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6394..6413) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7073..7933) /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 7696..7715 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7934..8038) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gcttaccatg 901 gcctacccct acgacgtgcc cgactacgcc tccctcggat ccgccaccat gacagctgac 961 aaggagaaga aaaggagtag ctcggagagg aggaaggaga agtcccggga tgctgcgcgg 1021 tgccggcgga gcaaggagac ggaggtgttc tatgagctgg cccatgagct gcctctgccc 1081 cacagtgtga gctcccatct ggacaaggcc tccatcatgc gactggcaat cagcttcctg 1141 cgaacacaca agctcctctc ctcagtttgc tctgaaaacg agtccgaagc cgaagctgac 1201 cagcagatgg acaacttgta cctgaaagcc ttggagggtt tcattgccgt ggtgacccaa 1261 gatggcgaca tgatctttct gtcagaaaac atcagcaagt tcatgggact tacacaggtg 1321 gagctaacag gacatagtat ctttgacttc actcatccct gcgaccatga ggagattcgt 1381 gagaacctga gtctcaaaaa tggctctggt tttgggaaaa aaagcaaaga catgtccaca 1441 gagcgggact tcttcatgag gatgaagtgc acggtcacca acagaggccg tactgtcaac 1501 ctcaagtcag ccacctggaa ggtcttgcac tgcacgggcc aggtgaaagt ctacaacaac 1561 tgccctcctc acaatagtct gtgtggctac aaggagcccc tgctgtcctg cctcatcatc 1621 atgtgtgaac caatccagca cccatcccac atggacatcc ccctggatag caagaccttc 1681 ctgagccgcc acagcatgga catgaagttc acctactgtg atgacagaat cacagaactg 1741 attggttacc accctgagga gctgcttggc cgctcagcct atgaattcta ccatgcgcta 1801 gactccgaga acatgaccaa gagtcaccag aacttgtgca ccaagggtca ggtagtaagt 1861 ggccagtacc ggatgctcgc aaagcatggg ggctacgtgt ggctggagac ccaggggacg 1921 gtcatctaca accctcgcaa cctgcagccc cagtgcatca tgtgtgtcaa ctacgtcctg 1981 agtgagattg agaagaatga cgtggtgttc tccatggacc agactgaatc cctgttcaag 2041 ccccacctga tggccatgaa cagcatcttt gatagcagtg gcaagggggc tgtgtctgag 2101 aagagtaact tcctattcac caagctaaag gaggagcccg aggagctggc ccagctggct 2161 cccaccccag gagacgccat catctctctg gatttcggga atcagaactt cgaggagtcc 2221 tcagcctatg gcaaggccat cctgcccccg agccagccat gggccacgga gttgaggagc 2281 cacagcaccc agagcgaggc tgggagcctg cctgccttca ccgtgcccca ggcagctgcc 2341 ccgggcagca ccacccccag tgccaccagc agcagcagca gctgctccac gcccaatagc 2401 cctgaagact attacacatc tttggataac gacctgaaga ttgaagtgat tgagaagctc 2461 ttcgccatgg acacagaggc caaggaccaa tgcagtaccc agacggattt caatgagctg 2521 gacttggaga cactggcacc ctatatcccc atggacgggg aagacttcca gctaagcccc 2581 atctgccccg aggagcggct cttggcggag aacccacagt ccacccccca gcactgcttc 2641 agtgccatga caaacatctt ccagccactg gcccctgtag ccccgcacag tcccttcctc 2701 ctggacaagt ttcagcagca gctggagagc aagaagacag agcccgagca ccggcccatg 2761 tcctccatct tctttgatgc cggaagcaaa gcatccctgc caccgtgctg tggccaggcc 2821 agcacccctc tctcttccat ggggggcaga tccaataccc agtggccccc agatccacca 2881 ttacattttg ggcccacaaa gtgggccgtc ggggatcagc gcacagagtt cttgggagca 2941 gcgccgttgg ggccccctgt ctctccaccc catgtctcca ccttcaagac aaggtctgca 3001 aagggttttg gggctcgagg cccagacgtg ctgagtccgg ccatggtagc cctctccaac 3061 aagctgaagc tgaagcgaca gctggagtat gaagagcaag ccttccagga cctgagcggg 3121 ggggacccac ctggtggcag cacctcacat ttgatgtgga aacggatgaa gaacctcagg 3181 ggtgggagct gccctttgat gccggacaag ccactgagcg caaatgtacc caatgataag 3241 ttcacccaaa accccatgag gggcctgggc catcccctga gacatctgcc gctgccacag 3301 cctccatctg ccatcagtcc cggggagaac agcaagagca ggttcccccc acagtgctac 3361 gccacccagt accaggacta cagcctgtcg tcagcccaca aggtgtcagg catggcaagc 3421 cggctgctcg ggccctcatt tgagtcctac ctgctgcccg aactgaccag atatgactgt 3481 gaggtgaacg tgcccgtgct gggaagctcc acgctcctgc aaggagggga cctcctcaga 3541 gccctggacc aggccacctg acaattctgc agatatccat cacactggcg gccgctcgag 3601 catgcatcta gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc 3661 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 3721 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 3781 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 3841 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 3901 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 3961 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 4021 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 4081 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 4141 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 4201 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 4261 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta 4321 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg 4381 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 4441 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 4501 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 4561 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 4621 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc 4681 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga 4741 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca 4801 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc 4861 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc 4921 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg 4981 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg 5041 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct 5101 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct 5161 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa 5221 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa 5281 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc 5341 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt 5401 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct 5461 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc 5521 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg 5581 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg 5641 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc 5701 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt 5761 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 5821 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt 5881 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 5941 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 6001 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 6061 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 6121 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 6181 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 6241 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 6301 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 6361 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 6421 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 6481 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 6541 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 6601 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 6661 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 6721 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 6781 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 6841 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 6901 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 6961 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 7021 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 7081 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 7141 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 7201 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 7261 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 7321 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 7381 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 7441 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 7501 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 7561 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 7621 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 7681 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 7741 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 7801 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 7861 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 7921 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 7981 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 8041 catttccccg aaaagtgcca cctgacgtc //