LOCUS Exported 7623 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS HA-HIF1alpha P402A/P564A-pBabe-puro SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7623) 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 7623) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7623 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 13..482 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 547..746 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 947..1363 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1293..1315 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1331..1347 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" CDS 1395..1421 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" regulatory 1434..1443 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(3958..3978) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3963..4292 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4143..4278 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4205..4224 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4302..4901 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(4302..4321) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4798..4818 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 5122..5591 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(5721..5738) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5892..6480) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5972..5991) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6640..7500) /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 7263..7282 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7501..7605) /gene="bla" /label=AmpR promoter ORIGIN 1 gccacctgca gcctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg 61 gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa 121 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc 181 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct 241 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc 301 cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact 361 gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt 421 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt 481 catttggggg ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg 541 ggaggcaagc tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgactga 601 ttttatgcgc ctgcgtcggt actagttagc taactagctc tgtatctggc ggacccgtgg 661 tggaactgac gagttctgaa cacccggccg caaccctggg agacgtccca gggactttgg 721 gggccgtttt tgtggcccga cctgaggaag ggagtcgatg tggaatccga ccccgtcagg 781 atatgtggtt ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt 841 gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct 901 gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc agactgttac 961 cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg ctcacaacca 1021 gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat ggccaacctt 1081 taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc aggttaagat 1141 caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca tcgtgacctg 1201 ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc ctaagcctcc 1261 gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc 1321 tcgatcctcc ctttatccag ccctcactcc ttctctaggc gccggccgga tcgaagacta 1381 agcttaccat ggcctacccc tacgacgtgc ccgactacgc ctccctcgga tccgccacca 1441 tggagggcgc cggcggcgcg aacgacaaga aaaagataag ttctgaacgt cgaaaagaaa 1501 agtctcgaga tgcagccaga tctcggcgaa gtaaagaatc tgaagttttt tatgagcttg 1561 ctcatcagtt gccacttcca cataatgtga gttcgcatct tgataaggcc tctgtgatga 1621 ggcttaccat cagctatttg cgtgtgagga aacttctgga tgctggtgat ttggatattg 1681 aagatgacat gaaagcacag atgaattgct tttatttgaa agccttggat ggttttgtta 1741 tggttctcac agatgatggt gacatgattt acatttctga taatgtgaac aaatacatgg 1801 gattaactca gtttgaacta actggacaca gtgtgtttga ttttactcat ccatgtgacc 1861 atgaggaaat gagagaaatg cttacacaca gaaatggcct tgtgaaaaag ggtaaagaac 1921 aaaacacaca gcgaagcttt tttctcagaa tgaagtgtac cctaactagc cgaggaagaa 1981 ctatgaacat aaagtctgca acatggaagg tattgcactg cacaggccac attcacgtat 2041 atgataccaa cagtaaccaa cctcagtgtg ggtataagaa accacctatg acctgcttgg 2101 tgctgatttg tgaacccatt cctcacccat caaatattga aattccttta gatagcaaga 2161 ctttcctcag tcgacacagc ctggatatga aattttctta ttgtgatgaa agaattaccg 2221 aattgatggg atatgagcca gaagaacttt taggccgctc aatttatgaa tattatcatg 2281 ctttggactc tgatcatctg accaaaactc atcatgatat gtttactaaa ggacaagtca 2341 ccacaggaca gtacaggatg cttgccaaaa gaggtggata tgtctgggtt gaaactcaag 2401 caactgtcat atataacacc aagaattctc aaccacagtg cattgtatgt gtgaattacg 2461 ttgtgagtgg tattattcag cacgacttga ttttctccct tcaacaaaca gaatgtgtcc 2521 ttaaaccggt tgaatcttca gatatgaaaa tgactcagct attcaccaaa gttgaatcag 2581 aagatacaag tagcctcttt gacaaactta agaaggaacc tgatgcttta actttgctgg 2641 ccgcagccgc tggagacaca atcatatctt tagattttgg cagcaacgac acagaaactg 2701 atgaccagca acttgaggaa gtaccattat ataatgatgt aatgctcccc tcacccaacg 2761 aaaaattaca gaatataaat ttggcaatgt ctccattacc caccgctgaa acgccaaagc 2821 cacttcgaag tagtgctgac cctgcactca atcaagaagt tgcattaaaa ttagaaccaa 2881 atccagagtc actggaactt tcttttacca tgccccagat tcaggatcag acacctagtc 2941 cttccgatgg aagcactaga caaagttcac ctgagcctaa tagtcccagt gaatattgtt 3001 tttatgtgga tagtgatatg gtcaatgaat tcaagttgga attggtagaa aaactttttg 3061 ctgaagacac agaagcaaag aacccatttt ctactcagga cacagattta gacttggaga 3121 tgttagctgc ttatatccca atggatgatg acttccagtt acgttccttc gatcagttgt 3181 caccattaga aagcagttcc gcaagccctg aaagcgcaag tcctcaaagc acagttacag 3241 tattccagca gactcaaata caagaaccta ctgctaatgc caccactacc actgccacca 3301 ctgatgaatt aaaaacagtg acaaaagacc gtatggaaga cattaaaata ttgattgcat 3361 ctccatctcc tacccacata cataaagaaa ctactagtgc cacatcatca ccatatagag 3421 atactcaaag tcggacagcc tcaccaaaca gagcaggaaa aggagtcata gaacagacag 3481 aaaaatctca tccaagaagc cctaacgtgt tatctgtcgc tttgagtcaa agaactacag 3541 ttcctgagga agaactaaat ccaaagatac tagctttgca gaatgctcag agaaagcgaa 3601 aaatggaaca tgatggttca ctttttcaag cagtaggaat tggaacatta ttacagcagc 3661 cagacgatca tgcagctact acatcacttt cttggaaacg tgtaaaagga tgcaaatcta 3721 gtgaacagaa tggaatggag caaaagacaa ttattttaat accctctgat ttagcatgta 3781 gactgctggg gcaatcaatg gatgaaagtg gattaccaca gctgaccagt tatgattgtg 3841 aagttaatgc tcctatacaa ggcagcagaa acctactgca gggtgaagaa ttactcagag 3901 ctttggatca agttaactga caattctgca gatatccatc acagtggtcg accctgtgga 3961 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 4021 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 4081 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 4141 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 4201 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag 4261 gaggcttttt tggaggccta ggcttttgca aaaagcttac catgaccgag tacaagccca 4321 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt 4381 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca 4441 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg 4501 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg 4561 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc 4621 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca 4681 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg 4741 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca 4801 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag 4861 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc cacgacccgc 4921 agcgcccgac cgaaaggagc gcacgacccc atgcatcgat aaaataaaag attttattta 4981 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag 5041 taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa 5101 ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt 5161 cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat 5221 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt 5281 ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga 5341 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg 5401 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc gccagtcctc 5461 cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg 5521 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg 5581 ggggtctttc acatgcagca tgtatcaaaa ttaatttggt tttttttctt aagtatttac 5641 attaaatggc catagttgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 5701 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 5761 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 5821 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 5881 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 5941 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 6001 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 6061 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 6121 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 6181 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 6241 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 6301 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg 6361 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 6421 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 6481 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 6541 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt gcggccgcaa 6601 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 6661 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 6721 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 6781 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 6841 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 6901 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 6961 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7021 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7081 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 7141 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 7201 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 7261 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 7321 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 7381 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 7441 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 7501 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 7561 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 7621 agt //