LOCUS Exported 6426 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Gateway Donor vector containing EGFR, part of the Target Accelerator Plasmid Collection.. ACCESSION . VERSION . KEYWORDS pDONR223_EGFR_p.Q1020H SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6426) 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 6426) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6426 /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..4303 /codon_start=1 /product="precursor of human epidermal growth factor receptor" /label=EGFR /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP DNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL PSPTDSNFYRALMDEEDMDDVVDADEYLIPHQGFFSSPSTSRTPLLSSLSATSNNSTVA CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA" protein_bind complement(4305..4404) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(4421..4440) /label=T7 /note="T7 promoter, forward primer" promoter complement(4422..4440) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4445..4461) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4445..4461) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 4892..5683 /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 5776..6364 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6265..6284 /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 atgccaactt tgtacaaaaa 661 agttggcacc atgcgaccct ccgggacggc cggggcagcg ctcctggcgc tgctggctgc 721 gctctgcccg gcgagtcggg ctctggagga aaagaaagtt tgccaaggca cgagtaacaa 781 gctcacgcag ttgggcactt ttgaagatca ttttctcagc ctccagagga tgttcaataa 841 ctgtgaggtg gtccttggga atttggaaat tacctatgtg cagaggaatt atgatctttc 901 cttcttaaag accatccagg aggtggctgg ttatgtcctc attgccctca acacagtgga 961 gcgaattcct ttggaaaacc tgcagatcat cagaggaaat atgtactacg aaaattccta 1021 tgccttagca gtcttatcta actatgatgc aaataaaacc ggactgaagg agctgcccat 1081 gagaaattta caggaaatcc tgcatggcgc cgtgcggttc agcaacaacc ctgccctgtg 1141 caacgtggag agcatccagt ggcgggacat agtcagcagt gactttctca gcaacatgtc 1201 gatggacttc cagaaccacc tgggcagctg ccaaaagtgt gatccaagct gtcccaatgg 1261 gagctgctgg ggtgcaggag aggagaactg ccagaaactg accaaaatca tctgtgccca 1321 gcagtgctcc gggcgctgcc gtggcaagtc ccccagtgac tgctgccaca accagtgtgc 1381 tgcaggctgc acaggccccc gggagagcga ctgcctggtc tgccgcaaat tccgagacga 1441 agccacgtgc aaggacacct gccccccact catgctctac aaccccacca cgtaccagat 1501 ggatgtgaac cccgagggca aatacagctt tggtgccacc tgcgtgaaga agtgtccccg 1561 taattatgtg gtgacagatc acggctcgtg cgtccgagcc tgtggggccg acagctatga 1621 gatggaggaa gacggcgtcc gcaagtgtaa gaagtgcgaa gggccttgcc gcaaagtgtg 1681 taacggaata ggtattggtg aatttaaaga ctcactctcc ataaatgcta cgaatattaa 1741 acacttcaaa aactgcacct ccatcagtgg cgatctccac atcctgccgg tggcatttag 1801 gggtgactcc ttcacacata ctcctcctct ggatccacag gaactggata ttctgaaaac 1861 cgtaaaggaa atcacagggt ttttgctgat tcaggcttgg cctgaaaaca ggacggacct 1921 ccatgccttt gagaacctag aaatcatacg cggcaggacc aagcaacatg gtcagttttc 1981 tcttgcagtc gtcagcctga acataacatc cttgggatta cgctccctca aggagataag 2041 tgatggagat gtgataattt caggaaacaa aaatttgtgc tatgcaaata caataaactg 2101 gaaaaaactg tttgggacct ccggtcagaa aaccaaaatt ataagcaaca gaggtgaaaa 2161 cagctgcaag gccacaggcc aggtctgcca tgccttgtgc tcccccgagg gctgctgggg 2221 cccggagccc agggactgcg tctcttgccg gaatgtcagc cgaggcaggg aatgcgtgga 2281 caagtgcaac cttctggagg gtgagccaag ggagtttgtg gagaactctg agtgcataca 2341 gtgccaccca gagtgcctgc ctcaggccat gaacatcacc tgcacaggac ggggaccaga 2401 caactgtatc cagtgtgccc actacattga cggcccccac tgcgtcaaga cctgcccggc 2461 aggagtcatg ggagaaaaca acaccctggt ctggaagtac gcagacgccg gccatgtgtg 2521 ccacctgtgc catccaaact gcacctacgg atgcactggg ccaggtcttg aaggctgtcc 2581 aacgaatggg cctaagatcc cgtccatcgc cactgggatg gtgggggccc tcctcttgct 2641 gctggtggtg gccctgggga tcggcctctt catgcgaagg cgccacatcg ttcggaagcg 2701 cacgctgcgg aggctgctgc aggagaggga gcttgtggag cctcttacac ccagtggaga 2761 agctcccaac caagctctct tgaggatctt gaaggaaact gaattcaaaa agatcaaagt 2821 gctgggctcc ggtgcgttcg gcacggtgta taagggactc tggatcccag aaggtgagaa 2881 agttaaaatt cccgtcgcta tcaaggaatt aagagaagca acatctccga aagccaacaa 2941 ggaaatcctc gatgaagcct acgtgatggc cagcgtggac aacccccacg tgtgccgcct 3001 gctgggcatc tgcctcacct ccaccgtgca actcatcacg cagctcatgc ccttcggctg 3061 cctcctggac tatgtccggg aacacaaaga caatattggc tcccagtacc tgctcaactg 3121 gtgtgtgcag atcgcaaagg gcatgaacta cttggaggac cgtcgcttgg tgcaccgcga 3181 cctggcagcc aggaacgtac tggtgaaaac accgcagcat gtcaagatca cagattttgg 3241 gctggccaaa ctgctgggtg cggaagagaa agaataccat gcagaaggag gcaaagtgcc 3301 tatcaagtgg atggcattgg aatcaatttt acacagaatc tatacccacc agagtgatgt 3361 ctggagctac ggggtgaccg tttgggagtt gatgaccttt ggatccaagc catatgacgg 3421 aatccctgcc agcgagatct cctccatcct ggagaaagga gaacgcctcc ctcagccacc 3481 catatgtacc atcgatgtct acatgatcat ggtcaagtgc tggatgatag acgcagatag 3541 tcgcccaaag ttccgtgagt tgatcatcga attctccaaa atggcccgag acccccagcg 3601 ctaccttgtc attcaggggg atgaaagaat gcatttgcca agtcctacag actccaactt 3661 ctaccgtgcc ctgatggatg aagaagacat ggacgacgtg gtggatgccg acgagtacct 3721 catcccacat cagggcttct tcagcagccc ctccacgtca cggactcccc tcctgagctc 3781 tctgagtgca accagcaaca attccaccgt ggcttgcatt gatagaaatg ggctgcaaag 3841 ctgtcccatc aaggaagaca gcttcttgca gcgatacagc tcagacccca caggcgcctt 3901 gactgaggac agcatagacg acaccttcct cccagtgcct gaatacataa accagtccgt 3961 tcccaaaagg cccgctggct ctgtgcagaa tcctgtctat cacaatcagc ctctgaaccc 4021 cgcgcccagc agagacccac actaccagga cccccacagc actgcagtgg gcaaccccga 4081 gtatctcaac actgtccagc ccacctgtgt caacagcaca ttcgacagcc ctgcccactg 4141 ggcccagaaa ggcagccacc aaattagcct ggacaaccct gactaccagc aggacttctt 4201 tcccaaggaa gccaagccaa atggcatctt taagggctcc acagctgaaa atgcagaata 4261 cctaagggtc gcgccacaaa gcagtgaatt tattggagca tgattgccaa ctttcttgta 4321 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat 4381 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta 4441 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat 4501 tgcacaagat aaaaatatat catcatgcct cctctagacc agccaggaca gaaatgcctc 4561 gacttcgctg ctacccaagg ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg 4621 aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc 4681 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt 4741 catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca 4801 agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca 4861 gggcagtcgc cctaaaacaa agttaaacat tatgagggaa gcggtgatcg ccgaagtatc 4921 gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc 4981 cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt 5041 gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct 5101 tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat 5161 tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg 5221 agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga 5281 tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc 5341 ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt 5401 aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt 5461 gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga 5521 ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc 5581 ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt 5641 ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga gccacccatg 5701 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa 5761 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa 5821 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc 5881 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta 5941 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct 6001 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg 6061 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag 6121 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc 6181 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg 6241 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt 6301 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg 6361 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca 6421 catgtt //