LOCUS       sequence-162226-        6432 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 6432)
  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 6432)
  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.774_775insHV
FEATURES             Location/Qualifiers
     source          1..6432
                     /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..3830)
                     /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
                     KANKEILDEAYVMASVDNPHVHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIG
                     SQYLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEY
                     HAEGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILE
                     KGERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERM
                     HLPSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNST
                     VACIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSV
                     QNPVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQ
                     ISLDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(3832..3931)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3948..3967)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3949..3967)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3972..3988)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3972..3988)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4419..5210
                     /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      5303..5891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5792..5811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6045..6062
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6221..6248)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6340..6426)
                     /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 gtccacgtgt
     2521 gccgcctgct gggcatctgc ctcacctcca ccgtgcaact catcacgcag ctcatgccct
     2581 tcggctgcct cctggactat gtccgggaac acaaagacaa tattggctcc cagtacctgc
     2641 tcaactggtg tgtgcagatc gcaaagggca tgaactactt ggaggaccgt cgcttggtgc
     2701 accgcgacct ggcagccagg aacgtactgg tgaaaacacc gcagcatgtc aagatcacag
     2761 attttgggct ggccaaactg ctgggtgcgg aagagaaaga ataccatgca gaaggaggca
     2821 aagtgcctat caagtggatg gcattggaat caattttaca cagaatctat acccaccaga
     2881 gtgatgtctg gagctacggg gtgaccgttt gggagttgat gacctttgga tccaagccat
     2941 atgacggaat ccctgccagc gagatctcct ccatcctgga gaaaggagaa cgcctccctc
     3001 agccacccat atgtaccatc gatgtctaca tgatcatggt caagtgctgg atgatagacg
     3061 cagatagtcg cccaaagttc cgtgagttga tcatcgaatt ctccaaaatg gcccgagacc
     3121 cccagcgcta ccttgtcatt cagggggatg aaagaatgca tttgccaagt cctacagact
     3181 ccaacttcta ccgtgccctg atggatgaag aagacatgga cgacgtggtg gatgccgacg
     3241 agtacctcat cccacagcag ggcttcttca gcagcccctc cacgtcacgg actcccctcc
     3301 tgagctctct gagtgcaacc agcaacaatt ccaccgtggc ttgcattgat agaaatgggc
     3361 tgcaaagctg tcccatcaag gaagacagct tcttgcagcg atacagctca gaccccacag
     3421 gcgccttgac tgaggacagc atagacgaca ccttcctccc agtgcctgaa tacataaacc
     3481 agtccgttcc caaaaggccc gctggctctg tgcagaatcc tgtctatcac aatcagcctc
     3541 tgaaccccgc gcccagcaga gacccacact accaggaccc ccacagcact gcagtgggca
     3601 accccgagta tctcaacact gtccagccca cctgtgtcaa cagcacattc gacagccctg
     3661 cccactgggc ccagaaaggc agccaccaaa ttagcctgga caaccctgac taccagcagg
     3721 acttctttcc caaggaagcc aagccaaatg gcatctttaa gggctccaca gctgaaaatg
     3781 cagaatacct aagggtcgcg ccacaaagca gtgaatttat tggagcatga ttgccaactt
     3841 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg
     3901 tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt
     3961 cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat
     4021 gttacattgc acaagataaa aatatatcat catgcctcct ctagaccagc caggacagaa
     4081 atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga
     4141 aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta
     4201 tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg
     4261 cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca
     4321 agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta
     4381 cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg
     4441 aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt
     4501 tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata
     4561 ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca
     4621 acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag
     4681 tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc
     4741 aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg
     4801 acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc
     4861 cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg
     4921 aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc
     4981 ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg
     5041 ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta
     5101 gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag
     5161 aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc
     5221 acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     5281 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5341 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5401 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5461 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5521 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5581 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5641 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5701 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5761 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5821 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5881 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5941 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6001 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6061 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     6121 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     6181 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat
     6241 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
     6301 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc
     6361 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt
     6421 tatttgatgc ct
//
