LOCUS       sequence-162208-        6435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing EGFR, part of the Target Accelerator
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6435)
  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 6435)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.ccell.2016.06.022"; journalName: "Cancer Cell"; date: 
            "2016-08-8- 8"; volume: "30"; issue: "2"; pages: "214-28"
COMMENT     Sequence Label: pDONR223_EGFR_p.769_770insASV
FEATURES             Location/Qualifiers
     source          1..6435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             join(192..263,264..2126,2127..2195,2196..3833)
                     /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
                     KANKEILDEAYVMASVASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNI
                     GSQYLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKE
                     YHAEGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSIL
                     EKGERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDER
                     MHLPSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNS
                     TVACIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGS
                     VQNPVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSH
                     QISLDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(3835..3934)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3951..3970)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3952..3970)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3975..3991)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3975..3991)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4422..5213
                     /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      5306..5894
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5795..5814
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6048..6065
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6224..6251)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6343..6429)
                     /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 ccagcgtggc cagcgtggac aacccccacg
     2521 tgtgccgcct gctgggcatc tgcctcacct ccaccgtgca actcatcacg cagctcatgc
     2581 ccttcggctg cctcctggac tatgtccggg aacacaaaga caatattggc tcccagtacc
     2641 tgctcaactg gtgtgtgcag atcgcaaagg gcatgaacta cttggaggac cgtcgcttgg
     2701 tgcaccgcga cctggcagcc aggaacgtac tggtgaaaac accgcagcat gtcaagatca
     2761 cagattttgg gctggccaaa ctgctgggtg cggaagagaa agaataccat gcagaaggag
     2821 gcaaagtgcc tatcaagtgg atggcattgg aatcaatttt acacagaatc tatacccacc
     2881 agagtgatgt ctggagctac ggggtgaccg tttgggagtt gatgaccttt ggatccaagc
     2941 catatgacgg aatccctgcc agcgagatct cctccatcct ggagaaagga gaacgcctcc
     3001 ctcagccacc catatgtacc atcgatgtct acatgatcat ggtcaagtgc tggatgatag
     3061 acgcagatag tcgcccaaag ttccgtgagt tgatcatcga attctccaaa atggcccgag
     3121 acccccagcg ctaccttgtc attcaggggg atgaaagaat gcatttgcca agtcctacag
     3181 actccaactt ctaccgtgcc ctgatggatg aagaagacat ggacgacgtg gtggatgccg
     3241 acgagtacct catcccacag cagggcttct tcagcagccc ctccacgtca cggactcccc
     3301 tcctgagctc tctgagtgca accagcaaca attccaccgt ggcttgcatt gatagaaatg
     3361 ggctgcaaag ctgtcccatc aaggaagaca gcttcttgca gcgatacagc tcagacccca
     3421 caggcgcctt gactgaggac agcatagacg acaccttcct cccagtgcct gaatacataa
     3481 accagtccgt tcccaaaagg cccgctggct ctgtgcagaa tcctgtctat cacaatcagc
     3541 ctctgaaccc cgcgcccagc agagacccac actaccagga cccccacagc actgcagtgg
     3601 gcaaccccga gtatctcaac actgtccagc ccacctgtgt caacagcaca ttcgacagcc
     3661 ctgcccactg ggcccagaaa ggcagccacc aaattagcct ggacaaccct gactaccagc
     3721 aggacttctt tcccaaggaa gccaagccaa atggcatctt taagggctcc acagctgaaa
     3781 atgcagaata cctaagggtc gcgccacaaa gcagtgaatt tattggagca tgattgccaa
     3841 ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac
     3901 aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg
     3961 agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct
     4021 gatgttacat tgcacaagat aaaaatatat catcatgcct cctctagacc agccaggaca
     4081 gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg acgcacaccg tggaaacgga
     4141 tgaaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc
     4201 gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag
     4261 tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat
     4321 ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg
     4381 ttacgcagca gggcagtcgc cctaaaacaa agttaaacat tatgagggaa gcggtgatcg
     4441 ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga
     4501 cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg
     4561 atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga
     4621 tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag
     4681 aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac
     4741 tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga
     4801 tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag
     4861 gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa
     4921 atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag
     4981 tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg
     5041 tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag
     5101 ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg
     5161 aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga
     5221 gccacccatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     5281 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5341 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5401 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5461 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5521 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5581 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5641 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5701 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5761 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5821 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5881 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5941 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6001 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     6061 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     6121 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     6181 ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag
     6241 gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc
     6301 tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag
     6361 agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg
     6421 ttttatttga tgcct
//
