LOCUS Exported 5854 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS HA-VHL W117R-pBabe-puro SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5854) AUTHORS Li L, Zhang L, Zhang X, Yan Q, Minamishima YA, Olumi AF, Mao M, Bartz S, Kaelin WG TITLE Hypoxia-inducible factor linked to differential kidney cancer risk seen with type 2A and type 2B VHL mutations. JOURNAL Mol Cell Biol. 2007 Aug . 27(15):5381-92. PUBMED 17526729 REFERENCE 2 (bases 1 to 5854) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5854 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 8..477 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 542..741 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 934..1350 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1318..1334 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" CDS 1379..1405 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(2093..2113) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2098..2427 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2278..2413 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2340..2359 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2437..3036 /codon_start=1 /gene="pac from Streptomyces" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA" primer_bind complement(2437..2456) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 2933..2953 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 3135..3728 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(3929..3946) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4100..4688) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4180..4199) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4866..5726) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 5489..5508 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5727..5831) /gene="bla" /label=AmpR promoter ORIGIN 1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca 61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt 121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag 181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc 241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga 301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc 361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc 421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt 481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg 541 taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta 601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa 661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc 721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg 781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt 841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt 901 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt 961 ttgaccttag atcactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc 1021 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg 1081 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca 1141 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct 1201 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttctt 1261 ccatccgcgc cgtctctccc ccttgaacct cctctttcga ccccgcctca atcctccctt 1321 tatccagccc tcactccttc tctaggcgcc ggccatcgag actaagctta ccatggccta 1381 cccctacgac gtgcccgact acgcctccct cggatccgcc accatgcccc ggagggcgga 1441 gaactgggac gaggccgagg taggcgcgga ggaggcaggc gtcgaagagt acggccctga 1501 agaagacggc ggggaggagt cgggcgccga ggagtccggc ccggaagagt ccggcccgga 1561 ggaactgggc gccgaggagg agatggaggc cgggcggccg cggcccgtgc tgcgctcggt 1621 gaactcgcgc gagccctccc aggtcatctt ctgcaatcgc agtccgcgcg tcgtgctgcc 1681 cgtatggctc aacttcgacg gcgagccgca gccctaccca acgctgccgc ctggcacggg 1741 ccgccgcatc cacagctacc gaggtcacct tcggctcttc agagatgcag ggacacacga 1801 tgggcttctg gttaaccaaa ctgaattatt tgtgccatct ctcaatgttg acggacagcc 1861 tatttttgcc aatatcacac tgccagtgta tactctgaaa gagcgatgcc tccaggttgt 1921 ccggagccta gtcaagcctg agaattacag gagactggac atcgtcaggt cgctctacga 1981 agatctggaa gaccacccaa atgtgcagaa agacctggag cggctgacac aggagcgcat 2041 tgcacatcaa cggatgggag attgaattcg ccagcacagt ggtcgaccct gtggaatgtg 2101 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 2161 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 2221 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 2281 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 2341 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc 2401 ttttttggag gcctaggctt ttgcaaaaag cttaccatga ccgagtacaa gcccacggtg 2461 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc 2521 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag 2581 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac 2641 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc 2701 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag 2761 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc 2821 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg 2881 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc 2941 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgagtgccc gaaggaccgc 3001 gcgacctggt gcatgacccg caagcccggt gcctgacgcc cgccccacga cccgcagcgc 3061 ccgaccgaaa ggagcgcacg accccatgca tcgataaaat aaaagatttt atttagtctc 3121 cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg 3181 ccattttgca aggcatggaa aaatacataa ctgagaatag agaagttcag atcaaggtca 3241 ggaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc 3301 cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg 3361 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc 3421 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga 3481 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct 3541 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt 3601 gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg 3661 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt 3721 ctttcatttc cgacttgtgg tctcgctgcc ttgggagggt ctcctctgag tgattgacta 3781 cccgtcagcg ggggtcttca catgcagcat gtatcaaaat taatttggtt ttttttctta 3841 agtatttaca ttaaatggcc atagttgcat taatgaatcg gccaacgcgc ggggagaggc 3901 ggtttgcgta ttggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 3961 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 4021 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 4081 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 4141 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 4201 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 4261 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 4321 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 4381 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 4441 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 4501 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg 4561 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 4621 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 4681 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 4741 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 4801 attaaaaatg aagtttgcgg ccgcaaatca atctaaagta tatatgagta aacttggtct 4861 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 4921 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 4981 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 5041 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 5101 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 5161 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 5221 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 5281 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 5341 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 5401 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 5461 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa 5521 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 5581 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 5641 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 5701 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 5761 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 5821 ggggttccgc gcacatttcc ccgaaaagtg ccac //