LOCUS       sequence-168748-        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.P622S
FEATURES             Location/Qualifiers
     source          1..6426
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             126..917
                     /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      1010..1598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1499..1518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1752..1769
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1928..1955)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2047..2133)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2182..2204
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2196..2213
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2197..2213
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2229..2328
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(2331..2402,2403..4265,4266..4334,4335..5963)
                     /codon_start=1
                     /product="precursor of human epidermal growth factor 
                     receptor"
                     /label=EGFR
                     /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE
                     DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL
                     QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ
                     WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR
                     CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP
                     EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI
                     GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK
                     EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD
                     GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG
                     PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP
                     DNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHSNCTYGCTGPGLEG
                     CPTNGPKIPSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP
                     SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(5965..6064)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6081..6100)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6082..6100)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6105..6121)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6105..6121)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt
       61 tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa
      121 acattatgag ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg
      181 tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg
      241 atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg
      301 atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag
      361 agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt
      421 ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg
      481 caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa
      541 gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg
      601 aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact
      661 gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa
      721 ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc
      781 agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct
      841 tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca
      901 aggtagtcgg caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg
      961 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1021 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1081 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1141 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     1201 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     1261 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     1321 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     1381 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     1441 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     1501 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     1561 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     1621 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     1681 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     1741 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     1801 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     1861 aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag
     1921 aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta
     1981 tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc
     2041 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
     2101 ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt
     2161 taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc agtcttaagc
     2221 tcgggcccca aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg
     2281 atgagcaatg cttttttata atgccaactt tgtacaaaaa agttggcacc atgcgaccct
     2341 ccgggacggc cggggcagcg ctcctggcgc tgctggctgc gctctgcccg gcgagtcggg
     2401 ctctggagga aaagaaagtt tgccaaggca cgagtaacaa gctcacgcag ttgggcactt
     2461 ttgaagatca ttttctcagc ctccagagga tgttcaataa ctgtgaggtg gtccttggga
     2521 atttggaaat tacctatgtg cagaggaatt atgatctttc cttcttaaag accatccagg
     2581 aggtggctgg ttatgtcctc attgccctca acacagtgga gcgaattcct ttggaaaacc
     2641 tgcagatcat cagaggaaat atgtactacg aaaattccta tgccttagca gtcttatcta
     2701 actatgatgc aaataaaacc ggactgaagg agctgcccat gagaaattta caggaaatcc
     2761 tgcatggcgc cgtgcggttc agcaacaacc ctgccctgtg caacgtggag agcatccagt
     2821 ggcgggacat agtcagcagt gactttctca gcaacatgtc gatggacttc cagaaccacc
     2881 tgggcagctg ccaaaagtgt gatccaagct gtcccaatgg gagctgctgg ggtgcaggag
     2941 aggagaactg ccagaaactg accaaaatca tctgtgccca gcagtgctcc gggcgctgcc
     3001 gtggcaagtc ccccagtgac tgctgccaca accagtgtgc tgcaggctgc acaggccccc
     3061 gggagagcga ctgcctggtc tgccgcaaat tccgagacga agccacgtgc aaggacacct
     3121 gccccccact catgctctac aaccccacca cgtaccagat ggatgtgaac cccgagggca
     3181 aatacagctt tggtgccacc tgcgtgaaga agtgtccccg taattatgtg gtgacagatc
     3241 acggctcgtg cgtccgagcc tgtggggccg acagctatga gatggaggaa gacggcgtcc
     3301 gcaagtgtaa gaagtgcgaa gggccttgcc gcaaagtgtg taacggaata ggtattggtg
     3361 aatttaaaga ctcactctcc ataaatgcta cgaatattaa acacttcaaa aactgcacct
     3421 ccatcagtgg cgatctccac atcctgccgg tggcatttag gggtgactcc ttcacacata
     3481 ctcctcctct ggatccacag gaactggata ttctgaaaac cgtaaaggaa atcacagggt
     3541 ttttgctgat tcaggcttgg cctgaaaaca ggacggacct ccatgccttt gagaacctag
     3601 aaatcatacg cggcaggacc aagcaacatg gtcagttttc tcttgcagtc gtcagcctga
     3661 acataacatc cttgggatta cgctccctca aggagataag tgatggagat gtgataattt
     3721 caggaaacaa aaatttgtgc tatgcaaata caataaactg gaaaaaactg tttgggacct
     3781 ccggtcagaa aaccaaaatt ataagcaaca gaggtgaaaa cagctgcaag gccacaggcc
     3841 aggtctgcca tgccttgtgc tcccccgagg gctgctgggg cccggagccc agggactgcg
     3901 tctcttgccg gaatgtcagc cgaggcaggg aatgcgtgga caagtgcaac cttctggagg
     3961 gtgagccaag ggagtttgtg gagaactctg agtgcataca gtgccaccca gagtgcctgc
     4021 ctcaggccat gaacatcacc tgcacaggac ggggaccaga caactgtatc cagtgtgccc
     4081 actacattga cggcccccac tgcgtcaaga cctgcccggc aggagtcatg ggagaaaaca
     4141 acaccctggt ctggaagtac gcagacgccg gccatgtgtg ccacctgtgc cattcaaact
     4201 gcacctacgg atgcactggg ccaggtcttg aaggctgtcc aacgaatggg cctaagatcc
     4261 cgtccatcgc cactgggatg gtgggggccc tcctcttgct gctggtggtg gccctgggga
     4321 tcggcctctt catgcgaagg cgccacatcg ttcggaagcg cacgctgcgg aggctgctgc
     4381 aggagaggga gcttgtggag cctcttacac ccagtggaga agctcccaac caagctctct
     4441 tgaggatctt gaaggaaact gaattcaaaa agatcaaagt gctgggctcc ggtgcgttcg
     4501 gcacggtgta taagggactc tggatcccag aaggtgagaa agttaaaatt cccgtcgcta
     4561 tcaaggaatt aagagaagca acatctccga aagccaacaa ggaaatcctc gatgaagcct
     4621 acgtgatggc cagcgtggac aacccccacg tgtgccgcct gctgggcatc tgcctcacct
     4681 ccaccgtgca actcatcacg cagctcatgc ccttcggctg cctcctggac tatgtccggg
     4741 aacacaaaga caatattggc tcccagtacc tgctcaactg gtgtgtgcag atcgcaaagg
     4801 gcatgaacta cttggaggac cgtcgcttgg tgcaccgcga cctggcagcc aggaacgtac
     4861 tggtgaaaac accgcagcat gtcaagatca cagattttgg gctggccaaa ctgctgggtg
     4921 cggaagagaa agaataccat gcagaaggag gcaaagtgcc tatcaagtgg atggcattgg
     4981 aatcaatttt acacagaatc tatacccacc agagtgatgt ctggagctac ggggtgaccg
     5041 tttgggagtt gatgaccttt ggatccaagc catatgacgg aatccctgcc agcgagatct
     5101 cctccatcct ggagaaagga gaacgcctcc ctcagccacc catatgtacc atcgatgtct
     5161 acatgatcat ggtcaagtgc tggatgatag acgcagatag tcgcccaaag ttccgtgagt
     5221 tgatcatcga attctccaaa atggcccgag acccccagcg ctaccttgtc attcaggggg
     5281 atgaaagaat gcatttgcca agtcctacag actccaactt ctaccgtgcc ctgatggatg
     5341 aagaagacat ggacgacgtg gtggatgccg acgagtacct catcccacag cagggcttct
     5401 tcagcagccc ctccacgtca cggactcccc tcctgagctc tctgagtgca accagcaaca
     5461 attccaccgt ggcttgcatt gatagaaatg ggctgcaaag ctgtcccatc aaggaagaca
     5521 gcttcttgca gcgatacagc tcagacccca caggcgcctt gactgaggac agcatagacg
     5581 acaccttcct cccagtgcct gaatacataa accagtccgt tcccaaaagg cccgctggct
     5641 ctgtgcagaa tcctgtctat cacaatcagc ctctgaaccc cgcgcccagc agagacccac
     5701 actaccagga cccccacagc actgcagtgg gcaaccccga gtatctcaac actgtccagc
     5761 ccacctgtgt caacagcaca ttcgacagcc ctgcccactg ggcccagaaa ggcagccacc
     5821 aaattagcct ggacaaccct gactaccagc aggacttctt tcccaaggaa gccaagccaa
     5881 atggcatctt taagggctcc acagctgaaa atgcagaata cctaagggtc gcgccacaaa
     5941 gcagtgaatt tattggagca tgattgccaa ctttcttgta caaagttggc attataagaa
     6001 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
     6061 tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct
     6121 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     6181 catcatgcct cctctagacc agccaggaca gaaatgcctc gacttcgctg ctacccaagg
     6241 ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct
     6301 gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct
     6361 tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt
     6421 tttttg
//
