LOCUS       sequence-168802-        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   Kim E, Ilic N, Shrestha Y, Zou L, Kamburov A, Zhu C, Yang X, Lubonja
            R, Tran N, Nguyen C, Lawrence MS, Piccioni F, Bagul M, Doench JG, 
            Chouinard CR, Wu X, Hogstrom L, Natoli T, Tamayo P, Horn H, Corsello
            SM, Lage K, Root DE, Subramanian A, Golub TR, Getz G, Boehm JS, Hahn
            WC
  TITLE     Systematic Functional Interrogation of Rare Cancer Variants 
            Identifies Oncogenic Alleles.
  JOURNAL   Cancer Discov. 2016 Jul;6(7):714-26. doi: 
            10.1158/2159-8290.CD-16-0160. Epub 2016 May 4.
  PUBMED    27147599
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: 
            "10.1158/2159-8290.CD-16-0160"; journalName: "Cancer Discov"; date: 
            "2016-07"; volume: "6"; issue: "7"; pages: "714-26"
COMMENT     Sequence Label: pDONR223_EGFR_p.A289V
FEATURES             Location/Qualifiers
     source          1..6426
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(671..742,743..2605,2606..2674,2675..4303)
                     /codon_start=1
                     /product="precursor of human epidermal growth factor 
                     receptor"
                     /label=EGFR
                     /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE
                     DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL
                     QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ
                     WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR
                     CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP
                     EGKYSFGVTCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI
                     GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK
                     EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD
                     GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG
                     PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP
                     DNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG
                     CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP
                     SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(4305..4404)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4421..4440)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4422..4440)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4445..4461)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4445..4461)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4892..5683
                     /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      5776..6364
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6265..6284
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcacc atgcgaccct ccgggacggc cggggcagcg ctcctggcgc tgctggctgc
      721 gctctgcccg gcgagtcggg ctctggagga aaagaaagtt tgccaaggca cgagtaacaa
      781 gctcacgcag ttgggcactt ttgaagatca ttttctcagc ctccagagga tgttcaataa
      841 ctgtgaggtg gtccttggga atttggaaat tacctatgtg cagaggaatt atgatctttc
      901 cttcttaaag accatccagg aggtggctgg ttatgtcctc attgccctca acacagtgga
      961 gcgaattcct ttggaaaacc tgcagatcat cagaggaaat atgtactacg aaaattccta
     1021 tgccttagca gtcttatcta actatgatgc aaataaaacc ggactgaagg agctgcccat
     1081 gagaaattta caggaaatcc tgcatggcgc cgtgcggttc agcaacaacc ctgccctgtg
     1141 caacgtggag agcatccagt ggcgggacat agtcagcagt gactttctca gcaacatgtc
     1201 gatggacttc cagaaccacc tgggcagctg ccaaaagtgt gatccaagct gtcccaatgg
     1261 gagctgctgg ggtgcaggag aggagaactg ccagaaactg accaaaatca tctgtgccca
     1321 gcagtgctcc gggcgctgcc gtggcaagtc ccccagtgac tgctgccaca accagtgtgc
     1381 tgcaggctgc acaggccccc gggagagcga ctgcctggtc tgccgcaaat tccgagacga
     1441 agccacgtgc aaggacacct gccccccact catgctctac aaccccacca cgtaccagat
     1501 ggatgtgaac cccgagggca aatacagctt tggtgtcacc tgcgtgaaga agtgtccccg
     1561 taattatgtg gtgacagatc acggctcgtg cgtccgagcc tgtggggccg acagctatga
     1621 gatggaggaa gacggcgtcc gcaagtgtaa gaagtgcgaa gggccttgcc gcaaagtgtg
     1681 taacggaata ggtattggtg aatttaaaga ctcactctcc ataaatgcta cgaatattaa
     1741 acacttcaaa aactgcacct ccatcagtgg cgatctccac atcctgccgg tggcatttag
     1801 gggtgactcc ttcacacata ctcctcctct ggatccacag gaactggata ttctgaaaac
     1861 cgtaaaggaa atcacagggt ttttgctgat tcaggcttgg cctgaaaaca ggacggacct
     1921 ccatgccttt gagaacctag aaatcatacg cggcaggacc aagcaacatg gtcagttttc
     1981 tcttgcagtc gtcagcctga acataacatc cttgggatta cgctccctca aggagataag
     2041 tgatggagat gtgataattt caggaaacaa aaatttgtgc tatgcaaata caataaactg
     2101 gaaaaaactg tttgggacct ccggtcagaa aaccaaaatt ataagcaaca gaggtgaaaa
     2161 cagctgcaag gccacaggcc aggtctgcca tgccttgtgc tcccccgagg gctgctgggg
     2221 cccggagccc agggactgcg tctcttgccg gaatgtcagc cgaggcaggg aatgcgtgga
     2281 caagtgcaac cttctggagg gtgagccaag ggagtttgtg gagaactctg agtgcataca
     2341 gtgccaccca gagtgcctgc ctcaggccat gaacatcacc tgcacaggac ggggaccaga
     2401 caactgtatc cagtgtgccc actacattga cggcccccac tgcgtcaaga cctgcccggc
     2461 aggagtcatg ggagaaaaca acaccctggt ctggaagtac gcagacgccg gccatgtgtg
     2521 ccacctgtgc catccaaact gcacctacgg atgcactggg ccaggtcttg aaggctgtcc
     2581 aacgaatggg cctaagatcc cgtccatcgc cactgggatg gtgggggccc tcctcttgct
     2641 gctggtggtg gccctgggga tcggcctctt catgcgaagg cgccacatcg ttcggaagcg
     2701 cacgctgcgg aggctgctgc aggagaggga gcttgtggag cctcttacac ccagtggaga
     2761 agctcccaac caagctctct tgaggatctt gaaggaaact gaattcaaaa agatcaaagt
     2821 gctgggctcc ggtgcgttcg gcacggtgta taagggactc tggatcccag aaggtgagaa
     2881 agttaaaatt cccgtcgcta tcaaggaatt aagagaagca acatctccga aagccaacaa
     2941 ggaaatcctc gatgaagcct acgtgatggc cagcgtggac aacccccacg tgtgccgcct
     3001 gctgggcatc tgcctcacct ccaccgtgca actcatcacg cagctcatgc ccttcggctg
     3061 cctcctggac tatgtccggg aacacaaaga caatattggc tcccagtacc tgctcaactg
     3121 gtgtgtgcag atcgcaaagg gcatgaacta cttggaggac cgtcgcttgg tgcaccgcga
     3181 cctggcagcc aggaacgtac tggtgaaaac accgcagcat gtcaagatca cagattttgg
     3241 gctggccaaa ctgctgggtg cggaagagaa agaataccat gcagaaggag gcaaagtgcc
     3301 tatcaagtgg atggcattgg aatcaatttt acacagaatc tatacccacc agagtgatgt
     3361 ctggagctac ggggtgaccg tttgggagtt gatgaccttt ggatccaagc catatgacgg
     3421 aatccctgcc agcgagatct cctccatcct ggagaaagga gaacgcctcc ctcagccacc
     3481 catatgtacc atcgatgtct acatgatcat ggtcaagtgc tggatgatag acgcagatag
     3541 tcgcccaaag ttccgtgagt tgatcatcga attctccaaa atggcccgag acccccagcg
     3601 ctaccttgtc attcaggggg atgaaagaat gcatttgcca agtcctacag actccaactt
     3661 ctaccgtgcc ctgatggatg aagaagacat ggacgacgtg gtggatgccg acgagtacct
     3721 catcccacag cagggcttct tcagcagccc ctccacgtca cggactcccc tcctgagctc
     3781 tctgagtgca accagcaaca attccaccgt ggcttgcatt gatagaaatg ggctgcaaag
     3841 ctgtcccatc aaggaagaca gcttcttgca gcgatacagc tcagacccca caggcgcctt
     3901 gactgaggac agcatagacg acaccttcct cccagtgcct gaatacataa accagtccgt
     3961 tcccaaaagg cccgctggct ctgtgcagaa tcctgtctat cacaatcagc ctctgaaccc
     4021 cgcgcccagc agagacccac actaccagga cccccacagc actgcagtgg gcaaccccga
     4081 gtatctcaac actgtccagc ccacctgtgt caacagcaca ttcgacagcc ctgcccactg
     4141 ggcccagaaa ggcagccacc aaattagcct ggacaaccct gactaccagc aggacttctt
     4201 tcccaaggaa gccaagccaa atggcatctt taagggctcc acagctgaaa atgcagaata
     4261 cctaagggtc gcgccacaaa gcagtgaatt tattggagca tgattgccaa ctttcttgta
     4321 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     4381 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     4441 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     4501 tgcacaagat aaaaatatat catcatgcct cctctagacc agccaggaca gaaatgcctc
     4561 gacttcgctg ctacccaagg ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg
     4621 aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc
     4681 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt
     4741 catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca
     4801 agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca
     4861 gggcagtcgc cctaaaacaa agttaaacat tatgagggaa gcggtgatcg ccgaagtatc
     4921 gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc
     4981 cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt
     5041 gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct
     5101 tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat
     5161 tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg
     5221 agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga
     5281 tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc
     5341 ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt
     5401 aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt
     5461 gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga
     5521 ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc
     5581 ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt
     5641 ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga gccacccatg
     5701 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     5761 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5821 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5881 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5941 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6001 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6061 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6121 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6181 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6241 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6301 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6361 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6421 catgtt
//
