LOCUS Exported 3360 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gateway Donor vector containing KRAS, part of the Target Accelerator Plasmid Collection.. ACCESSION . VERSION . KEYWORDS pDONR223_KRAS_p.G13V SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3360) AUTHORS Berger AH, Brooks AN, Wu X, Shrestha Y, Chouinard C, Piccioni F, Bagul M, Kamburov A, Imielinski M, Hogstrom L, Zhu C, Yang X, Pantel S, Sakai R, Watson J, Kaplan N, Campbell JD, Singh S, Root DE, Narayan R, Natoli T, Lahr DL, Tirosh I, Tamayo P, Getz G, Wong B, Doench J, Subramanian A, Golub TR, Meyerson M, Boehm JS TITLE High-throughput Phenotyping of Lung Cancer Somatic Mutations. JOURNAL Cancer Cell. 2016 Aug 8;30(2):214-28. doi: 10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28. PUBMED 27478040 REFERENCE 2 (bases 1 to 3360) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3360 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 92..109 /label=L4440 /note="L4440 vector, forward primer" terminator 268..295 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 387..473 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 522..544 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 536..553 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 537..553 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 569..668 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 671..1237 /codon_start=1 /product="human small GTPase proto-oncoprotein" /label=K-Ras /translation="MTEYKLVVVGAGVVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI RKHKEKMSKDGKKKKKKSKTKCVIM" protein_bind complement(1239..1338) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1355..1374) /label=T7 /note="T7 promoter, forward primer" promoter complement(1356..1374) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1379..1395) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1379..1395) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 1826..2617 /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 2710..3298 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3199..3218 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc 241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta 301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc 361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa 421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg 481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa 541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac 601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgacaactt tgtacaaaaa 661 agttggcacc atgactgaat ataaacttgt ggtagttgga gctggtgttg taggcaagag 721 tgccttgacg atacagctaa ttcagaatca ttttgtggac gaatatgatc caacaataga 781 ggattcctac aggaagcaag tagtaattga tggagaaacc tgtctcttgg atattctcga 841 cacagcaggt caagaggagt acagtgcaat gagggaccag tacatgagga ctggggaggg 901 ctttctttgt gtatttgcca taaataatac taaatcattt gaagatattc accattatag 961 agaacaaatt aaaagagtta aggactctga agatgtacct atggtcctag taggaaataa 1021 atgtgatttg ccttctagaa cagtagacac aaaacaggct caggacttag caagaagtta 1081 tggaattcct tttattgaaa catcagcaaa gacaagacag ggtgttgatg atgccttcta 1141 tacattagtt cgagaaattc gaaaacataa agaaaagatg agcaaagatg gtaaaaagaa 1201 gaaaaagaag tcaaagacaa agtgtgtaat tatgtaattg ccaactttct tgtacaaagt 1261 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca 1321 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt 1381 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca 1441 agataaaaat atatcatcat gcctcctcta gaccagccag gacagaaatg cctcgacttc 1501 gctgctaccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca 1561 gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa 1621 ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc 1681 ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg 1741 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag 1801 tcgccctaaa acaaagttaa acattatgag ggaagcggtg atcgccgaag tatcgactca 1861 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca 1921 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt 1981 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga 2041 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt 2101 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg 2161 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc 2221 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga 2281 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct 2341 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg 2401 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc 2461 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct 2521 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta 2581 cgtgaaaggc gagatcacca aggtagtcgg caaataaccc tcgagccacc catgaccaaa 2641 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa 2701 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc 2761 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 2821 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg 2881 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 2941 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 3001 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 3061 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 3121 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 3181 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 3241 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 3301 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt //