LOCUS Exported 3435 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway Donor vector containing VHL, part of the Target Accelerator Plasmid Collection.. ACCESSION . VERSION . KEYWORDS pDONR223_VHL_p.S111N SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3435) AUTHORS Kim E, Ilic N, Shrestha Y, Zou L, Kamburov A, Zhu C, Yang X, Lubonja R, Tran N, Nguyen C, Lawrence MS, Piccioni F, Bagul M, Doench JG, Chouinard CR, Wu X, Hogstrom L, Natoli T, Tamayo P, Horn H, Corsello SM, Lage K, Root DE, Subramanian A, Golub TR, Getz G, Boehm JS, Hahn WC TITLE Systematic Functional Interrogation of Rare Cancer Variants Identifies Oncogenic Alleles. JOURNAL Cancer Discov. 2016 Jul;6(7):714-26. doi: 10.1158/2159-8290.CD-16-0160. Epub 2016 May 4. PUBMED 27147599 REFERENCE 2 (bases 1 to 3435) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3435 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 12..600 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 501..520 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 754..771 /label=L4440 /note="L4440 vector, forward primer" terminator 930..957 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 1049..1135 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 1184..1206 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 1198..1215 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1199..1215 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 1231..1330 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(1976..2075) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(2092..2111) /label=T7 /note="T7 promoter, forward primer" promoter complement(2093..2111) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2116..2132) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2116..2132) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 2563..3354 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" ORIGIN 1 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 61 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 121 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta 181 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 241 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 301 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 361 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 421 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 481 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 541 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 601 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 661 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 721 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa 781 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg 841 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc 901 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct 961 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct 1021 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca 1081 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc 1141 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt 1201 aaaacgacgg ccagtcttaa gctcgggccc caaataatga ttttattttg actgatagtg 1261 acctgttcgt tgcaacaaat tgatgagcaa tgctttttta taatgccaac tttgtacaaa 1321 aaagttggca ccatgccccg gagggcggag aactgggacg aggccgaggt aggcgcggag 1381 gaggcaggcg tcgaagagta cggccctgaa gaagacggcg gggaggagtc gggcgccgag 1441 gagtccggcc cggaagagtc cggcccggag gaactgggcg ccgaggagga gatggaggcc 1501 gggcggccgc ggcccgtgct gcgctcggtg aactcgcgcg agccctccca ggtcatcttc 1561 tgcaatcgca gtccgcgcgt cgtgctgccc gtatggctca acttcgacgg cgagccgcag 1621 ccctacccaa cgctgccgcc tggcacgggc cgccgcatcc acaactaccg aggtcacctt 1681 tggctcttca gagatgcagg gacacacgat gggcttctgg ttaaccaaac tgaattattt 1741 gtgccatctc tcaatgttga cggacagcct atttttgcca atatcacact gccagtgtat 1801 actctgaaag agcgatgcct ccaggttgtc cggagcctag tcaagcctga gaattacagg 1861 agactggaca tcgtcaggtc gctctacgaa gatctggaag accacccaaa tgtgcagaaa 1921 gacctggagc ggctgacaca ggagcgcatt gcacatcaac ggatgggaga ttgattgcca 1981 actttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa 2041 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt 2101 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc 2161 tgatgttaca ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac 2221 agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg 2281 atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag 2341 cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca 2401 gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca 2461 tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat 2521 gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc 2581 gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg 2641 acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt 2701 gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg 2761 atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta 2821 gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa 2881 ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg 2941 atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta 3001 ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta 3061 aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta 3121 gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat 3181 gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa 3241 gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg 3301 gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg 3361 agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac 3421 cccgtagaaa agatc //