LOCUS Exported 8070 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS HA-HIF2alpha-pcDNA3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8070) 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 8070) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8070 /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..4708 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4559..4694 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4621..4640 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4775..5569 /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(4829..4848) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5439..5458 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5743..5864 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5780..5799) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5834..5853 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5913..5929) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5913..5929) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5926..5948) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5937..5953 /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(5961..5991) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6006..6027 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6144..6161) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6315..6903) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6395..6414) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7074..7934) /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 7697..7716 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7935..8039) /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 gcttaccatg 901 gcctacccnt acgacgtgcc cgactacgcc tccctcggat ccgccaccat gacagctgac 961 aaggagaaga aaaggagtag ctcggagagg aggaaggaga agtcccggga tgctgcgcgg 1021 tgccggcgga gcaaggagac ggaggtgttc tatgagctgg cccatgagct gcctctgccc 1081 cacagtgtga gctcccatct ggacaaggcc tccatcatgc gactggaaat 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 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 4141 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 4201 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 4261 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta 4321 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg 4381 tgtcagttag ggtgtggaaa gtccccaggc tccccaggca ggcagaagta tgcaaagcat 4441 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag 4501 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat 4561 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt 4621 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg 4681 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg 4741 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc 4801 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 4861 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 4921 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg 4981 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 5041 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 5101 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 5161 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 5221 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 5281 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg 5341 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 5401 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 5461 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 5521 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg 5581 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 5641 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 5701 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct 5761 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 5821 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg 5881 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt 5941 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt 6001 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg 6061 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg 6121 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 6181 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 6241 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 6301 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 6361 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 6421 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 6481 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg 6541 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 6601 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 6661 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 6721 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg 6781 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 6841 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 6901 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 6961 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 7021 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 7081 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 7141 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 7201 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 7261 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 7321 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 7381 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 7441 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 7501 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 7561 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 7621 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 7681 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 7741 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 7801 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 7861 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 7921 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 7981 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 8041 acatttcccc gaaaagtgcc acctgacgtc //