LOCUS Exported 4609 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Gateway Donor vector containing ARAF, part of the Target Accelerator Plasmid Collection.. ACCESSION . VERSION . KEYWORDS pDONR223_ARAF_p.D429A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4609) 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 4609) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4609 /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" protein_bind complement(2009..2108) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(2125..2144) /label=T7 /note="T7 promoter, forward primer" promoter complement(2126..2144) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2149..2165) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2149..2165) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 2596..3387 /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 3480..4068 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3969..3988 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4222..4239 /label=L4440 /note="L4440 vector, forward primer" terminator 4398..4425 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 4517..4603 /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 aatgccaact ttgtacaaaa 181 aagttggcat ggagccacca cggggccccc ctgccaatgg ggccgagcca tcccgggcag 241 tgggcaccgt caaagtatac ctgcccaaca agcaacgcac ggtggtgact gtccgggatg 301 gcatgagtgt ctacgactct ctagacaagg ccctgaaggt gcggggtcta aatcaggact 361 gctgtgtggt ctaccgactc atcaagggac gaaagacggt cactgcctgg gacacagcca 421 ttgctcccct ggatggcgag gagctcattg tcgaggtcct tgaagatgtc ccgctgacca 481 tgcacaattt tgtacggaag accttcttca gcctggcgtt ctgtgacttc tgccttaagt 541 ttctgttcca tggcttccgt tgccaaacct gtggctacaa gttccaccag cattgttcct 601 ccaaggtccc cacagtctgt gttgacatga gtaccaaccg ccaacagttc taccacagtg 661 tccaggattt gtccggaggc tccagacagc atgaggctcc ctcgaaccgc cccctgaatg 721 agttgctaac cccccagggt cccagccccc gcacccagca ctgtgacccg gagcacttcc 781 ccttccctgc cccagccaat gcccccctac agcgcatccg ctccacgtcc actcccaacg 841 tccatatggt cagcaccacg gcccccatgg actccaacct catccagctc actggccaga 901 gtttcagcac tgatgctgcc ggtagtagag gaggtagtga tggaaccccc cgggggagcc 961 ccagcccagc cagcgtgtcc tcggggagga agtccccaca ttccaagtca ccagcagagc 1021 agcgcgagcg gaagtccttg gccgatgaca agaagaaagt gaagaacctg gggtaccggg 1081 actcaggcta ttactgggag gtaccaccca gtgaggtgca gctgctgaag aggatcggga 1141 cgggctcgtt tggcaccgtg tttcgagggc ggtggcatgg cgatgtggcc gtgaaggtgc 1201 tcaaggtgtc ccagcccaca gctgagcagg cccaggcttt caagaatgag atgcaggtgc 1261 tcaggaagac gcgacatgtc aacatcttgc tgtttatggg cttcatgacc cggccgggat 1321 ttgccatcat cacacagtgg tgtgagggct ccagcctcta ccatcacctg catgtggccg 1381 acacacgctt cgacatggtc cagctcatcg acgtggcccg gcagactgcc cagggcatgg 1441 actacctcca tgccaagaac atcatccacc gagctctcaa gtctaacaac atcttcctac 1501 atgaggggct cacggtgaag atcggtgact ttggcttggc cacagtgaag actcgatgga 1561 gcggggccca gcccttggag cagccctcag gatctgtgct gtggatggca gctgaggtga 1621 tccgtatgca ggacccgaac ccctacagct tccagtcaga cgtctatgcc tacggggttg 1681 tgctctacga gcttatgact ggctcactgc cttacagcca cattggctgc cgtgaccaga 1741 ttatctttat ggtgggccgt ggctatctgt ccccggacct cagcaaaatc tccagcaact 1801 gccccaaggc catgcggcgc ctgctgtctg actgcctcaa gttccagcgg gaggagcggc 1861 ccctcttccc ccagatcctg gccacaattg agctgctgca acggtcactc cccaagattg 1921 agcggagtgc ctcggaaccc tccttgcacc gcacccaggc cgatgagttg cctgcctgcc 1981 tactcagcgc agcccgcctt gtgccttagc ccaactttct tgtacaaagt tggcattata 2041 agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc 2101 attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt 2161 tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat 2221 atatcatcat gcctcctcta gaccagccag gacagaaatg cctcgacttc gctgctaccc 2281 aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca gtggacataa 2341 gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga 2401 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac 2461 tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg 2521 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa 2581 acaaagttaa acattatgag ggaagcggtg atcgccgaag tatcgactca actatcagag 2641 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc 2701 tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc 2761 gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct 2821 tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac 2881 atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat 2941 gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg 3001 acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat 3061 ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg 3121 ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac 3181 agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc 3241 ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa 3301 gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc 3361 gagatcacca aggtagtcgg caaataaccc tcgagccacc catgaccaaa atcccttaac 3421 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct 3481 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 3541 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc 3601 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc 3661 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct 3721 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag 3781 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc 3841 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg 3901 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag 3961 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt 4021 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac 4081 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg 4141 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc 4201 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata 4261 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt 4321 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa 4381 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc 4441 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca 4501 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa 4561 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcct //