LOCUS Exported 5851 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS HA-VHL Y98N-pBabe-puro SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5851) 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 5851) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5851 /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(2090..2110) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2095..2424 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2275..2410 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2337..2356 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2434..3033 /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(2434..2453) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 2930..2950 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 3132..3725 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(3926..3943) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4097..4685) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4177..4196) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4863..5723) /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 5486..5505 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5724..5828) /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 cggatccacc atgccccgga gggcggagaa 1441 ctgggacgag gccgaggtag gcgcggagga ggcaggcgtc gaagagtacg gccctgaaga 1501 agacggcggg gaggagtcgg gcgccgagga gtccggcccg gaagagtccg gcccggagga 1561 actgggcgcc gaggaggaga tggaggccgg gcggccgcgg cccgtgctgc gctcggtgaa 1621 ctcgcgcgag ccctcccagg tcatcttctg caatcgcagt ccgcgcgtcg tgctgcccgt 1681 atggctcaac ttcgacggcg agccgcagcc caacccaacg ctgccgcctg gcacgggccg 1741 ccgcatccac agctaccgag gtcacctttg gctcttcaga gatgcaggga cacacgatgg 1801 gcttctggtt aaccaaactg aattatttgt gccatctctc aatgttgacg gacagcctat 1861 ttttgccaat atcacactgc cagtgtatac tctgaaagag cgatgcctcc aggttgtccg 1921 gagcctagtc aagcctgaga attacaggag actggacatc gtcaggtcgc tctacgaaga 1981 tctggaagac cacccaaatg tgcagaaaga cctggagcgg ctgacacagg agcgcattgc 2041 acatcaacgg atgggagatt gaattcgcca gcacagtggt cgaccctgtg gaatgtgtgt 2101 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 2161 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg 2221 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg 2281 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt 2341 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt 2401 tttggaggcc taggcttttg caaaaagctt accatgaccg agtacaagcc cacggtgcgc 2461 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac 2521 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg 2581 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac 2641 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc 2701 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg 2761 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc 2821 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag 2881 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc 2941 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg agtgcccgaa ggaccgcgcg 3001 acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg 3061 accgaaagga gcgcacgacc ccatgcatcg ataaaataaa agattttatt tagtctccag 3121 aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca 3181 ttttgcaagg catggaaaaa tacataactg agaatagaga agttcagatc aaggtcagga 3241 acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc 3301 ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat atctgtggta 3361 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtccagccct 3421 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc 3481 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct 3541 ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc tccgattgac 3601 tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg 3661 tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt 3721 tcatttccga cttgtggtct cgctgccttg ggagggtctc ctctgagtga ttgactaccc 3781 gtcagcgggg gtcttcacat gcagcatgta tcaaaattaa tttggttttt tttcttaagt 3841 atttacatta aatggccata gttgcattaa tgaatcggcc aacgcgcggg gagaggcggt 3901 ttgcgtattg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 3961 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 4021 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 4081 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 4141 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 4201 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 4261 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 4321 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 4381 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 4441 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 4501 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc 4561 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 4621 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 4681 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 4741 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 4801 aaaaatgaag tttgcggccg caaatcaatc taaagtatat atgagtaaac ttggtctgac 4861 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 4921 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 4981 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 5041 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 5101 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 5161 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 5221 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 5281 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 5341 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 5401 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 5461 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg 5521 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 5581 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 5641 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 5701 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 5761 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 5821 gttccgcgca catttccccg aaaagtgcca c //