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.K754E 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 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" CDS 192..3824 /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 EANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA" protein_bind complement(3826..3925) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(3942..3961) /label=T7 /note="T7 promoter, forward primer" promoter complement(3943..3961) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3966..3982) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3966..3982) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 4413..5204 /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 5297..5885 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5786..5805 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6039..6056 /label=L4440 /note="L4440 vector, forward primer" terminator 6215..6242 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 6334..6420 /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 aagttggcac catgcgaccc tccgggacgg ccggggcagc gctcctggcg ctgctggctg 241 cgctctgccc ggcgagtcgg gctctggagg aaaagaaagt ttgccaaggc acgagtaaca 301 agctcacgca gttgggcact tttgaagatc attttctcag cctccagagg atgttcaata 361 actgtgaggt ggtccttggg aatttggaaa ttacctatgt gcagaggaat tatgatcttt 421 ccttcttaaa gaccatccag gaggtggctg gttatgtcct cattgccctc aacacagtgg 481 agcgaattcc tttggaaaac ctgcagatca tcagaggaaa tatgtactac gaaaattcct 541 atgccttagc agtcttatct aactatgatg caaataaaac cggactgaag gagctgccca 601 tgagaaattt acaggaaatc ctgcatggcg ccgtgcggtt cagcaacaac cctgccctgt 661 gcaacgtgga gagcatccag tggcgggaca tagtcagcag tgactttctc agcaacatgt 721 cgatggactt ccagaaccac ctgggcagct gccaaaagtg tgatccaagc tgtcccaatg 781 ggagctgctg gggtgcagga gaggagaact gccagaaact gaccaaaatc atctgtgccc 841 agcagtgctc cgggcgctgc cgtggcaagt cccccagtga ctgctgccac aaccagtgtg 901 ctgcaggctg cacaggcccc cgggagagcg actgcctggt ctgccgcaaa ttccgagacg 961 aagccacgtg caaggacacc tgccccccac tcatgctcta caaccccacc acgtaccaga 1021 tggatgtgaa ccccgagggc aaatacagct ttggtgccac ctgcgtgaag aagtgtcccc 1081 gtaattatgt ggtgacagat cacggctcgt gcgtccgagc ctgtggggcc gacagctatg 1141 agatggagga agacggcgtc cgcaagtgta agaagtgcga agggccttgc cgcaaagtgt 1201 gtaacggaat aggtattggt gaatttaaag actcactctc cataaatgct acgaatatta 1261 aacacttcaa aaactgcacc tccatcagtg gcgatctcca catcctgccg gtggcattta 1321 ggggtgactc cttcacacat actcctcctc tggatccaca ggaactggat attctgaaaa 1381 ccgtaaagga aatcacaggg tttttgctga ttcaggcttg gcctgaaaac aggacggacc 1441 tccatgcctt tgagaaccta gaaatcatac gcggcaggac caagcaacat ggtcagtttt 1501 ctcttgcagt cgtcagcctg aacataacat ccttgggatt acgctccctc aaggagataa 1561 gtgatggaga tgtgataatt tcaggaaaca aaaatttgtg ctatgcaaat acaataaact 1621 ggaaaaaact gtttgggacc tccggtcaga aaaccaaaat tataagcaac agaggtgaaa 1681 acagctgcaa ggccacaggc caggtctgcc atgccttgtg ctcccccgag ggctgctggg 1741 gcccggagcc cagggactgc gtctcttgcc ggaatgtcag ccgaggcagg gaatgcgtgg 1801 acaagtgcaa ccttctggag ggtgagccaa gggagtttgt ggagaactct gagtgcatac 1861 agtgccaccc agagtgcctg cctcaggcca tgaacatcac ctgcacagga cggggaccag 1921 acaactgtat ccagtgtgcc cactacattg acggccccca ctgcgtcaag acctgcccgg 1981 caggagtcat gggagaaaac aacaccctgg tctggaagta cgcagacgcc ggccatgtgt 2041 gccacctgtg ccatccaaac tgcacctacg gatgcactgg gccaggtctt gaaggctgtc 2101 caacgaatgg gcctaagatc ccgtccatcg ccactgggat ggtgggggcc ctcctcttgc 2161 tgctggtggt ggccctgggg atcggcctct tcatgcgaag gcgccacatc gttcggaagc 2221 gcacgctgcg gaggctgctg caggagaggg agcttgtgga gcctcttaca cccagtggag 2281 aagctcccaa ccaagctctc ttgaggatct tgaaggaaac tgaattcaaa aagatcaaag 2341 tgctgggctc cggtgcgttc ggcacggtgt ataagggact ctggatccca gaaggtgaga 2401 aagttaaaat tcccgtcgct atcaaggaat taagagaagc aacatctccg gaagccaaca 2461 aggaaatcct cgatgaagcc tacgtgatgg ccagcgtgga caacccccac gtgtgccgcc 2521 tgctgggcat ctgcctcacc tccaccgtgc aactcatcac gcagctcatg cccttcggct 2581 gcctcctgga ctatgtccgg gaacacaaag acaatattgg ctcccagtac ctgctcaact 2641 ggtgtgtgca gatcgcaaag ggcatgaact acttggagga ccgtcgcttg gtgcaccgcg 2701 acctggcagc caggaacgta ctggtgaaaa caccgcagca tgtcaagatc acagattttg 2761 ggctggccaa actgctgggt gcggaagaga aagaatacca tgcagaagga ggcaaagtgc 2821 ctatcaagtg gatggcattg gaatcaattt tacacagaat ctatacccac cagagtgatg 2881 tctggagcta cggggtgacc gtttgggagt tgatgacctt tggatccaag ccatatgacg 2941 gaatccctgc cagcgagatc tcctccatcc tggagaaagg agaacgcctc cctcagccac 3001 ccatatgtac catcgatgtc tacatgatca tggtcaagtg ctggatgata gacgcagata 3061 gtcgcccaaa gttccgtgag ttgatcatcg aattctccaa aatggcccga gacccccagc 3121 gctaccttgt cattcagggg gatgaaagaa tgcatttgcc aagtcctaca gactccaact 3181 tctaccgtgc cctgatggat gaagaagaca tggacgacgt ggtggatgcc gacgagtacc 3241 tcatcccaca gcagggcttc ttcagcagcc cctccacgtc acggactccc ctcctgagct 3301 ctctgagtgc aaccagcaac aattccaccg tggcttgcat tgatagaaat gggctgcaaa 3361 gctgtcccat caaggaagac agcttcttgc agcgatacag ctcagacccc acaggcgcct 3421 tgactgagga cagcatagac gacaccttcc tcccagtgcc tgaatacata aaccagtccg 3481 ttcccaaaag gcccgctggc tctgtgcaga atcctgtcta tcacaatcag cctctgaacc 3541 ccgcgcccag cagagaccca cactaccagg acccccacag cactgcagtg ggcaaccccg 3601 agtatctcaa cactgtccag cccacctgtg tcaacagcac attcgacagc cctgcccact 3661 gggcccagaa aggcagccac caaattagcc tggacaaccc tgactaccag caggacttct 3721 ttcccaagga agccaagcca aatggcatct ttaagggctc cacagctgaa aatgcagaat 3781 acctaagggt cgcgccacaa agcagtgaat ttattggagc atgattgcca actttcttgt 3841 acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta 3901 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt 3961 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca 4021 ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac agaaatgcct 4081 cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac 4141 gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct 4201 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt 4261 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc 4321 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc 4381 agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat 4441 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg 4501 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt 4561 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc 4621 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca 4681 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg 4741 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg 4801 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg 4861 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct 4921 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt 4981 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg 5041 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg 5101 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg 5161 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat 5221 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa 5281 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 5341 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 5401 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt 5461 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 5521 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 5581 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 5641 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 5701 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 5761 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 5821 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 5881 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc 5941 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt 6001 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag 6061 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca 6121 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc 6181 gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt 6241 ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg 6301 ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac 6361 cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg 6421 atgcct //