LOCUS Exported 3263 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway ORF Entry clone of human HRAS [NM_005343.2 ] with stop codon (for native or N-terminal fusions), Q61K mutation. ACCESSION . VERSION . KEYWORDS Hs.HRAS Q61K SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3263) AUTHORS Esposito TITLE RAS mutant collection V2.0 JOURNAL unpublished REFERENCE 2 (bases 1 to 3263) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3263 /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" CDS 186..755 /codon_start=1 /product="human small GTPase proto-oncoprotein" /label=H-Ras /translation="MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV IDGETCLLDILDTAGKEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVK DSDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREI RQHKLRKLNPPDESGPGCMSCKCVLS" protein_bind complement(754..853) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 1230..2021 /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 2114..2702 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2603..2622 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2856..2873 /label=L4440 /note="L4440 vector, forward primer" terminator 3032..3059 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 3151..3237 /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 gcaccatgac ggaatataag ctggtggtgg tgggcgccgg cggtgtgggc aagagtgcgc 241 tgaccatcca gctgatccag aaccattttg tggacgaata cgaccccact atagaggatt 301 cctaccggaa gcaggtggtc attgatgggg agacgtgcct gttggacatc ctggataccg 361 ccggcaagga ggagtacagc gccatgcggg accagtacat gcgcaccggg gagggcttcc 421 tgtgtgtgtt tgccatcaac aacaccaagt cttttgagga catccaccag tacagggagc 481 agatcaaacg ggtgaaggac tcggatgacg tgcccatggt gctggtgggg aacaagtgtg 541 acctggctgc acgcactgtg gaatctcggc aggctcagga cctcgcccga agctacggca 601 tcccctacat cgagacctcg gccaagaccc ggcagggagt ggaggatgcc ttctacacgt 661 tggtgcgtga gatccggcag cacaagctgc ggaagctgaa ccctcctgat gagagtggcc 721 ccggctgcat gagctgcaag tgtgtgctct cctaatcaac tttcttgtac aaagttggca 781 ttataggaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata 841 aaatcattat ttgcatgcaa gcttagatcc tctagaccag ccaggacaga aatgcctcga 901 cttcgctgct acccaaggtt gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa 961 cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac 1021 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca 1081 tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag 1141 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg 1201 gcagtcgccc taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc gaagtatcga 1261 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg 1321 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc 1381 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt 1441 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg 1501 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag 1561 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc 1621 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg 1681 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa 1741 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt 1801 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact 1861 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt 1921 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc 1981 actacgtgaa aggcgagatc accaaggtag tcggcaaata accctcgagc cacccatgac 2041 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga 2101 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 2161 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 2221 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt 2281 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 2341 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 2401 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 2461 tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca 2521 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 2581 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 2641 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 2701 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 2761 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag 2821 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg 2881 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct 2941 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta 3001 ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg 3061 cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg 3121 cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga 3181 caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg 3241 cctggcagtt ccctactctc gcg //