LOCUS Exported 3284 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway ORF Entry clone of human NRAS [NM_002524.4 ] with stop codon (for native or N-terminal fusions), G12D mutation. ACCESSION . VERSION . KEYWORDS Hs.NRAS G12D SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3284) AUTHORS Esposito TITLE RAS mutant collection V2.0 JOURNAL unpublished REFERENCE 2 (bases 1 to 3284) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3284 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 23..45 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 37..54 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 38..54 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 84..183 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(775..874) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 1251..2042 /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 2135..2723 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2624..2643 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2877..2894 /label=L4440 /note="L4440 vector, forward primer" terminator 3053..3080 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 3172..3258 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" ORIGIN 1 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaaa 61 attcgagctc ggtccgagga tccgaaataa tgattttatt ttgactgata gtgacctgtt 121 cgttgcaaca aattgataag caatgctttt ttataatgcc aactttgtac aaaaaagttg 181 gcaggagaga tggctctgcc gccaccatga ctgagtacaa actggtggtg gttggagcag 241 atggtgttgg gaaaagcgca ctgacaatcc agctaatcca gaaccacttt gtagatgaat 301 atgatcccac catagaggat tcttacagaa aacaagtggt tatagatggt gaaacctgtt 361 tgttggacat actggataca gctggacaag aagagtacag tgccatgaga gaccaataca 421 tgaggacagg cgaaggcttc ctctgtgtat ttgccatcaa taatagcaag tcatttgcgg 481 atattaacct ctacagggag cagattaagc gagtaaaaga ctcggatgat gtacctatgg 541 tgctagtggg aaacaagtgt gatttgccaa caaggacagt tgatacaaaa caagcccacg 601 aactggccaa gagttacggg attccattca ttgaaacctc agccaagacc agacagggtg 661 ttgaagatgc tttttacaca ctggtaagag aaatacgcca gtaccgaatg aaaaaactca 721 acagcagtga tgatgggact cagggttgta tgggattgcc atgtgtggtg atgtaatcaa 781 ctttcttgta caaagttggc attataggaa agcattgctt atcaatttgt tgcaacgaac 841 aggtcactat cagtcaaaat aaaatcatta tttgcatgca agcttagatc ctctagacca 901 gccaggacag aaatgcctcg acttcgctgc tacccaaggt tgccgggtga cgcacaccgt 961 ggaaacggat gaaggcacga acccagtgga cataagcctg ttcggttcgt aagctgtaat 1021 gcaagtagcg tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta 1081 acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttggggta cagtctatgc 1141 ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca 1201 gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag 1261 cggtgatcgc cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc 1321 tcgaaccgac gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc 1381 cacacagtga tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc 1441 gagctttgat caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc 1501 gcgctgtaga agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta 1561 agcgcgaact gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc 1621 cagccacgat cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg 1681 ccttggtagg tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg 1741 aggcgctaaa tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc 1801 gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc 1861 cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca 1921 tacttgaagc tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag 1981 atcagttgga agaatttgtc cactacgtga aaggcgagat caccaaggta gtcggcaaat 2041 aaccctcgag ccacccatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag 2101 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa 2161 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag 2221 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg 2281 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat 2341 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta 2401 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg 2461 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc 2521 gtgagcattg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa 2581 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc 2641 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt 2701 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct 2761 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc 2821 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg 2881 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt 2941 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc 3001 gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc 3061 atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc 3121 ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct 3181 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg 3241 agcctttcgt tttatttgat gcctggcagt tccctactct cgcg //