LOCUS       sequence-161712-        6426 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 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 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.K754E
FEATURES             Location/Qualifiers
     source          1..6426
                     /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..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      complement(6215..6242)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(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
//
