LOCUS Exported 6429 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.773_774insH SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6429) 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 6429) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6429 /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..3827 /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 KANKEILDEAYVMASVDNPHHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGS QYLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYH AEGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEK GERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMH LPSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTV ACIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQ NPVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQI SLDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA" protein_bind complement(3829..3928) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(3945..3964) /label=T7 /note="T7 promoter, forward primer" promoter complement(3946..3964) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3969..3985) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3969..3985) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 4416..5207 /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 5300..5888 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5789..5808 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6042..6059 /label=L4440 /note="L4440 vector, forward primer" terminator 6218..6245 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 6337..6423 /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 aaagccaaca 2461 aggaaatcct cgatgaagcc tacgtgatgg ccagcgtgga caacccccac cacgtgtgcc 2521 gcctgctggg catctgcctc acctccaccg tgcaactcat cacgcagctc atgcccttcg 2581 gctgcctcct ggactatgtc cgggaacaca aagacaatat tggctcccag tacctgctca 2641 actggtgtgt gcagatcgca aagggcatga actacttgga ggaccgtcgc ttggtgcacc 2701 gcgacctggc agccaggaac gtactggtga aaacaccgca gcatgtcaag atcacagatt 2761 ttgggctggc caaactgctg ggtgcggaag agaaagaata ccatgcagaa ggaggcaaag 2821 tgcctatcaa gtggatggca ttggaatcaa ttttacacag aatctatacc caccagagtg 2881 atgtctggag ctacggggtg accgtttggg agttgatgac ctttggatcc aagccatatg 2941 acggaatccc tgccagcgag atctcctcca tcctggagaa aggagaacgc ctccctcagc 3001 cacccatatg taccatcgat gtctacatga tcatggtcaa gtgctggatg atagacgcag 3061 atagtcgccc aaagttccgt gagttgatca tcgaattctc caaaatggcc cgagaccccc 3121 agcgctacct tgtcattcag ggggatgaaa gaatgcattt gccaagtcct acagactcca 3181 acttctaccg tgccctgatg gatgaagaag acatggacga cgtggtggat gccgacgagt 3241 acctcatccc acagcagggc ttcttcagca gcccctccac gtcacggact cccctcctga 3301 gctctctgag tgcaaccagc aacaattcca ccgtggcttg cattgataga aatgggctgc 3361 aaagctgtcc catcaaggaa gacagcttct tgcagcgata cagctcagac cccacaggcg 3421 ccttgactga ggacagcata gacgacacct tcctcccagt gcctgaatac ataaaccagt 3481 ccgttcccaa aaggcccgct ggctctgtgc agaatcctgt ctatcacaat cagcctctga 3541 accccgcgcc cagcagagac ccacactacc aggaccccca cagcactgca gtgggcaacc 3601 ccgagtatct caacactgtc cagcccacct gtgtcaacag cacattcgac agccctgccc 3661 actgggccca gaaaggcagc caccaaatta gcctggacaa ccctgactac cagcaggact 3721 tctttcccaa ggaagccaag ccaaatggca tctttaaggg ctccacagct gaaaatgcag 3781 aatacctaag ggtcgcgcca caaagcagtg aatttattgg agcatgattg ccaactttct 3841 tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca 3901 ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt 3961 attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt 4021 acattgcaca agataaaaat atatcatcat gcctcctcta gaccagccag gacagaaatg 4081 cctcgacttc gctgctaccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg 4141 cacgaaccca gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc 4201 gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg 4261 ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc 4321 agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc 4381 agcagggcag tcgccctaaa acaaagttaa acattatgag ggaagcggtg atcgccgaag 4441 tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc 4501 tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg 4561 atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg 4621 accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca 4681 ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat 4741 ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca 4801 ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag 4861 cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa 4921 ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta 4981 cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg 5041 ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac 5101 aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat 5161 ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataaccc tcgagccacc 5221 catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag 5281 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 5341 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 5401 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc 5461 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 5521 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 5581 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 5641 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 5701 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 5761 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 5821 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 5881 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 5941 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg 6001 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg 6061 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat 6121 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata 6181 cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc 6241 cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg 6301 ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt 6361 caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat 6421 ttgatgcct //