LOCUS Exported 3372 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway Donor vector containing RHOA , part of the Target Accelerator Plasmid Collection.. ACCESSION . VERSION . KEYWORDS pDONR223_RHOA_p.Y42C SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3372) 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 3372) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3372 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 43..65 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 57..74 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 58..74 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 90..189 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 192..770 /codon_start=1 /product="human Rho family small GTPase RhoA" /label=RhoA /translation="MAAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENCVAD IEVDGKQVELALWDTAGQEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVK HFCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKT KDGVREVFEMATRAALQARRGKKKSGCLVL" protein_bind complement(772..871) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(888..907) /label=T7 /note="T7 promoter, forward primer" promoter complement(889..907) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(912..928) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(912..928) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 1359..2150 /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" rep_origin 2243..2831 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2732..2751 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2985..3002 /label=L4440 /note="L4440 vector, forward primer" terminator 3161..3188 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 3280..3366 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" ORIGIN 1 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta 61 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga 121 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgacaact ttgtacaaaa 181 aagttggcac catggctgcc atccggaaga aactggtgat tgttggtgat ggagcctgtg 241 gaaagacatg cttgctcata gtcttcagca aggaccagtt cccagaggtg tatgtgccca 301 cagtgtttga gaactgtgtg gcagatatcg aggtggatgg aaagcaggta gagttggctt 361 tgtgggacac agctgggcag gaagattatg atcgcctgag gcccctctcc tacccagata 421 ccgatgttat actgatgtgt ttttccatcg acagccctga tagtttagaa aacatcccag 481 aaaagtggac cccagaagtc aagcatttct gtcccaacgt gcccatcatc ctggttggga 541 ataagaagga tcttcggaat gatgagcaca caaggcggga gctagccaag atgaagcagg 601 agccggtgaa acctgaagaa ggcagagata tggcaaacag gattggcgct tttgggtaca 661 tggagtgttc agcaaagacc aaagatggag tgagagaggt ttttgaaatg gctacgagag 721 ctgctctgca agctagacgt gggaagaaaa aatctgggtg ccttgtcttg tgcccaactt 781 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg 841 tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt 901 cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat 961 gttacattgc acaagataaa aatatatcat catgcctcct ctagaccagc caggacagaa 1021 atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga 1081 aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta 1141 tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg 1201 cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca 1261 agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta 1321 cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg 1381 aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt 1441 tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata 1501 ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca 1561 acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag 1621 tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc 1681 aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg 1741 acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc 1801 cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg 1861 aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc 1921 ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg 1981 ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta 2041 gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag 2101 aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc 2161 acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg 2221 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 2281 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 2341 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt 2401 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 2461 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 2521 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 2581 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 2641 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 2701 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 2761 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 2821 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 2881 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 2941 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 3001 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 3061 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta 3121 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat 3181 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc 3241 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc 3301 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt 3361 tatttgatgc ct //