LOCUS       sequence-162231-        6405 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 6405)
  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 6405)
  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.ELREA_746del
FEATURES             Location/Qualifiers
     source          1..6405
                     /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(189..260,261..2123,2124..2192,2193..3806)
                     /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
                     SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKTSPKANKE
                     ILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQYLLNW
                     CVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHAEGGKV
                     PIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKGERLPQ
                     PPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHLPSPTD
                     SNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVACIDRN
                     GLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQNPVYHN
                     QPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQISLDNPD
                     YQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(3805..3904)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3921..3940)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3922..3940)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3945..3961)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3945..3961)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4392..5183
                     /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      5276..5864
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5765..5784
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6018..6035
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6194..6221)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6313..6399)
                     /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 gcgaccctcc gggacggccg gggcagcgct cctggcgctg ctggctgcgc
      241 tctgcccggc gagtcgggct ctggaggaaa agaaagtttg ccaaggcacg agtaacaagc
      301 tcacgcagtt gggcactttt gaagatcatt ttctcagcct ccagaggatg ttcaataact
      361 gtgaggtggt ccttgggaat ttggaaatta cctatgtgca gaggaattat gatctttcct
      421 tcttaaagac catccaggag gtggctggtt atgtcctcat tgccctcaac acagtggagc
      481 gaattccttt ggaaaacctg cagatcatca gaggaaatat gtactacgaa aattcctatg
      541 ccttagcagt cttatctaac tatgatgcaa ataaaaccgg actgaaggag ctgcccatga
      601 gaaatttaca ggaaatcctg catggcgccg tgcggttcag caacaaccct gccctgtgca
      661 acgtggagag catccagtgg cgggacatag tcagcagtga ctttctcagc aacatgtcga
      721 tggacttcca gaaccacctg ggcagctgcc aaaagtgtga tccaagctgt cccaatggga
      781 gctgctgggg tgcaggagag gagaactgcc agaaactgac caaaatcatc tgtgcccagc
      841 agtgctccgg gcgctgccgt ggcaagtccc ccagtgactg ctgccacaac cagtgtgctg
      901 caggctgcac aggcccccgg gagagcgact gcctggtctg ccgcaaattc cgagacgaag
      961 ccacgtgcaa ggacacctgc cccccactca tgctctacaa ccccaccacg taccagatgg
     1021 atgtgaaccc cgagggcaaa tacagctttg gtgccacctg cgtgaagaag tgtccccgta
     1081 attatgtggt gacagatcac ggctcgtgcg tccgagcctg tggggccgac agctatgaga
     1141 tggaggaaga cggcgtccgc aagtgtaaga agtgcgaagg gccttgccgc aaagtgtgta
     1201 acggaatagg tattggtgaa tttaaagact cactctccat aaatgctacg aatattaaac
     1261 acttcaaaaa ctgcacctcc atcagtggcg atctccacat cctgccggtg gcatttaggg
     1321 gtgactcctt cacacatact cctcctctgg atccacagga actggatatt ctgaaaaccg
     1381 taaaggaaat cacagggttt ttgctgattc aggcttggcc tgaaaacagg acggacctcc
     1441 atgcctttga gaacctagaa atcatacgcg gcaggaccaa gcaacatggt cagttttctc
     1501 ttgcagtcgt cagcctgaac ataacatcct tgggattacg ctccctcaag gagataagtg
     1561 atggagatgt gataatttca ggaaacaaaa atttgtgcta tgcaaataca ataaactgga
     1621 aaaaactgtt tgggacctcc ggtcagaaaa ccaaaattat aagcaacaga ggtgaaaaca
     1681 gctgcaaggc cacaggccag gtctgccatg ccttgtgctc ccccgagggc tgctggggcc
     1741 cggagcccag ggactgcgtc tcttgccgga atgtcagccg aggcagggaa tgcgtggaca
     1801 agtgcaacct tctggagggt gagccaaggg agtttgtgga gaactctgag tgcatacagt
     1861 gccacccaga gtgcctgcct caggccatga acatcacctg cacaggacgg ggaccagaca
     1921 actgtatcca gtgtgcccac tacattgacg gcccccactg cgtcaagacc tgcccggcag
     1981 gagtcatggg agaaaacaac accctggtct ggaagtacgc agacgccggc catgtgtgcc
     2041 acctgtgcca tccaaactgc acctacggat gcactgggcc aggtcttgaa ggctgtccaa
     2101 cgaatgggcc taagatcccg tccatcgcca ctgggatggt gggggccctc ctcttgctgc
     2161 tggtggtggc cctggggatc ggcctcttca tgcgaaggcg ccacatcgtt cggaagcgca
     2221 cgctgcggag gctgctgcag gagagggagc ttgtggagcc tcttacaccc agtggagaag
     2281 ctcccaacca agctctcttg aggatcttga aggaaactga attcaaaaag atcaaagtgc
     2341 tgggctccgg tgcgttcggc acggtgtata agggactctg gatcccagaa ggtgagaaag
     2401 ttaaaattcc cgtcgctatc aaaacatctc cgaaagccaa caaggaaatc ctcgatgaag
     2461 cctacgtgat ggccagcgtg gacaaccccc acgtgtgccg cctgctgggc atctgcctca
     2521 cctccaccgt gcaactcatc acgcagctca tgcccttcgg ctgcctcctg gactatgtcc
     2581 gggaacacaa agacaatatt ggctcccagt acctgctcaa ctggtgtgtg cagatcgcaa
     2641 agggcatgaa ctacttggag gaccgtcgct tggtgcaccg cgacctggca gccaggaacg
     2701 tactggtgaa aacaccgcag catgtcaaga tcacagattt tgggctggcc aaactgctgg
     2761 gtgcggaaga gaaagaatac catgcagaag gaggcaaagt gcctatcaag tggatggcat
     2821 tggaatcaat tttacacaga atctataccc accagagtga tgtctggagc tacggggtga
     2881 ccgtttggga gttgatgacc tttggatcca agccatatga cggaatccct gccagcgaga
     2941 tctcctccat cctggagaaa ggagaacgcc tccctcagcc acccatatgt accatcgatg
     3001 tctacatgat catggtcaag tgctggatga tagacgcaga tagtcgccca aagttccgtg
     3061 agttgatcat cgaattctcc aaaatggccc gagaccccca gcgctacctt gtcattcagg
     3121 gggatgaaag aatgcatttg ccaagtccta cagactccaa cttctaccgt gccctgatgg
     3181 atgaagaaga catggacgac gtggtggatg ccgacgagta cctcatccca cagcagggct
     3241 tcttcagcag cccctccacg tcacggactc ccctcctgag ctctctgagt gcaaccagca
     3301 acaattccac cgtggcttgc attgatagaa atgggctgca aagctgtccc atcaaggaag
     3361 acagcttctt gcagcgatac agctcagacc ccacaggcgc cttgactgag gacagcatag
     3421 acgacacctt cctcccagtg cctgaataca taaaccagtc cgttcccaaa aggcccgctg
     3481 gctctgtgca gaatcctgtc tatcacaatc agcctctgaa ccccgcgccc agcagagacc
     3541 cacactacca ggacccccac agcactgcag tgggcaaccc cgagtatctc aacactgtcc
     3601 agcccacctg tgtcaacagc acattcgaca gccctgccca ctgggcccag aaaggcagcc
     3661 accaaattag cctggacaac cctgactacc agcaggactt ctttcccaag gaagccaagc
     3721 caaatggcat ctttaagggc tccacagctg aaaatgcaga atacctaagg gtcgcgccac
     3781 aaagcagtga atttattgga gcatgaccaa ctttcttgta caaagttggc attataagaa
     3841 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
     3901 tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct
     3961 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     4021 catcatgcct cctctagacc agccaggaca gaaatgcctc gacttcgctg ctacccaagg
     4081 ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct
     4141 gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct
     4201 tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt
     4261 tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc
     4321 gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa
     4381 agttaaacat tatgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag
     4441 ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg
     4501 cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa
     4561 ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc
     4621 ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca
     4681 ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca
     4741 ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa
     4801 aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg
     4861 ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc
     4921 ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg
     4981 cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc
     5041 cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag
     5101 atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga
     5161 tcaccaaggt agtcggcaaa taaccctcga gccacccatg accaaaatcc cttaacgtga
     5221 gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5281 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5341 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5401 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5461 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5521 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5581 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5641 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5701 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5761 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5821 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5881 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5941 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6001 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     6061 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     6121 actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc caggaagagt
     6181 ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc
     6241 agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc
     6301 cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
     6361 aaaggcccag tcttccgact gagcctttcg ttttatttga tgcct
//
