LOCUS       sequence-473542-        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_WT
FEATURES             Location/Qualifiers
     source          1..6426
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1663..1762)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1779..1798)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1780..1798)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1803..1819)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1803..1819)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2250..3041
                     /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      3134..3722
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3623..3642
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3876..3893
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4052..4079)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4171..4257)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4306..4328
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4320..4337
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4321..4337
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4353..4452
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(4455..4526,4527..6389,6390..6426,1..32,33..1661)
                     /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
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
ORIGIN
        1 tggtggtggc cctggggatc ggcctcttca tgcgaaggcg ccacatcgtt cggaagcgca
       61 cgctgcggag gctgctgcag gagagggagc ttgtggagcc tcttacaccc agtggagaag
      121 ctcccaacca agctctcttg aggatcttga aggaaactga attcaaaaag atcaaagtgc
      181 tgggctccgg tgcgttcggc acggtgtata agggactctg gatcccagaa ggtgagaaag
      241 ttaaaattcc cgtcgctatc aaggaattaa gagaagcaac atctccgaaa gccaacaagg
      301 aaatcctcga tgaagcctac gtgatggcca gcgtggacaa cccccacgtg tgccgcctgc
      361 tgggcatctg cctcacctcc accgtgcaac tcatcacgca gctcatgccc ttcggctgcc
      421 tcctggacta tgtccgggaa cacaaagaca atattggctc ccagtacctg ctcaactggt
      481 gtgtgcagat cgcaaagggc atgaactact tggaggaccg tcgcttggtg caccgcgacc
      541 tggcagccag gaacgtactg gtgaaaacac cgcagcatgt caagatcaca gattttgggc
      601 tggccaaact gctgggtgcg gaagagaaag aataccatgc agaaggaggc aaagtgccta
      661 tcaagtggat ggcattggaa tcaattttac acagaatcta tacccaccag agtgatgtct
      721 ggagctacgg ggtgaccgtt tgggagttga tgacctttgg atccaagcca tatgacggaa
      781 tccctgccag cgagatctcc tccatcctgg agaaaggaga acgcctccct cagccaccca
      841 tatgtaccat cgatgtctac atgatcatgg tcaagtgctg gatgatagac gcagatagtc
      901 gcccaaagtt ccgtgagttg atcatcgaat tctccaaaat ggcccgagac ccccagcgct
      961 accttgtcat tcagggggat gaaagaatgc atttgccaag tcctacagac tccaacttct
     1021 accgtgccct gatggatgaa gaagacatgg acgacgtggt ggatgccgac gagtacctca
     1081 tcccacagca gggcttcttc agcagcccct ccacgtcacg gactcccctc ctgagctctc
     1141 tgagtgcaac cagcaacaat tccaccgtgg cttgcattga tagaaatggg ctgcaaagct
     1201 gtcccatcaa ggaagacagc ttcttgcagc gatacagctc agaccccaca ggcgccttga
     1261 ctgaggacag catagacgac accttcctcc cagtgcctga atacataaac cagtccgttc
     1321 ccaaaaggcc cgctggctct gtgcagaatc ctgtctatca caatcagcct ctgaaccccg
     1381 cgcccagcag agacccacac taccaggacc cccacagcac tgcagtgggc aaccccgagt
     1441 atctcaacac tgtccagccc acctgtgtca acagcacatt cgacagccct gcccactggg
     1501 cccagaaagg cagccaccaa attagcctgg acaaccctga ctaccagcag gacttctttc
     1561 ccaaggaagc caagccaaat ggcatcttta agggctccac agctgaaaat gcagaatacc
     1621 taagggtcgc gccacaaagc agtgaattta ttggagcatg attgccaact ttcttgtaca
     1681 aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca
     1741 gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca
     1801 tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg
     1861 cacaagataa aaatatatca tcatgcctcc tctagaccag ccaggacaga aatgcctcga
     1921 cttcgctgct acccaaggtt gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa
     1981 cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac
     2041 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
     2101 tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag
     2161 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg
     2221 gcagtcgccc taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc gaagtatcga
     2281 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
     2341 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc
     2401 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt
     2461 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
     2521 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
     2581 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc
     2641 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
     2701 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
     2761 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
     2821 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
     2881 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt
     2941 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc
     3001 actacgtgaa aggcgagatc accaaggtag tcggcaaata accctcgagc cacccatgac
     3061 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga
     3121 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3181 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3241 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     3301 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3361 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3421 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3481 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3541 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3601 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3661 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3721 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3781 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3841 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3901 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     3961 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta
     4021 ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg
     4081 cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg
     4141 cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga
     4201 caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg
     4261 cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt
     4321 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
     4381 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
     4441 aaaaagttgg caccatgcga ccctccggga cggccggggc agcgctcctg gcgctgctgg
     4501 ctgcgctctg cccggcgagt cgggctctgg aggaaaagaa agtttgccaa ggcacgagta
     4561 acaagctcac gcagttgggc acttttgaag atcattttct cagcctccag aggatgttca
     4621 ataactgtga ggtggtcctt gggaatttgg aaattaccta tgtgcagagg aattatgatc
     4681 tttccttctt aaagaccatc caggaggtgg ctggttatgt cctcattgcc ctcaacacag
     4741 tggagcgaat tcctttggaa aacctgcaga tcatcagagg aaatatgtac tacgaaaatt
     4801 cctatgcctt agcagtctta tctaactatg atgcaaataa aaccggactg aaggagctgc
     4861 ccatgagaaa tttacaggaa atcctgcatg gcgccgtgcg gttcagcaac aaccctgccc
     4921 tgtgcaacgt ggagagcatc cagtggcggg acatagtcag cagtgacttt ctcagcaaca
     4981 tgtcgatgga cttccagaac cacctgggca gctgccaaaa gtgtgatcca agctgtccca
     5041 atgggagctg ctggggtgca ggagaggaga actgccagaa actgaccaaa atcatctgtg
     5101 cccagcagtg ctccgggcgc tgccgtggca agtcccccag tgactgctgc cacaaccagt
     5161 gtgctgcagg ctgcacaggc ccccgggaga gcgactgcct ggtctgccgc aaattccgag
     5221 acgaagccac gtgcaaggac acctgccccc cactcatgct ctacaacccc accacgtacc
     5281 agatggatgt gaaccccgag ggcaaataca gctttggtgc cacctgcgtg aagaagtgtc
     5341 cccgtaatta tgtggtgaca gatcacggct cgtgcgtccg agcctgtggg gccgacagct
     5401 atgagatgga ggaagacggc gtccgcaagt gtaagaagtg cgaagggcct tgccgcaaag
     5461 tgtgtaacgg aataggtatt ggtgaattta aagactcact ctccataaat gctacgaata
     5521 ttaaacactt caaaaactgc acctccatca gtggcgatct ccacatcctg ccggtggcat
     5581 ttaggggtga ctccttcaca catactcctc ctctggatcc acaggaactg gatattctga
     5641 aaaccgtaaa ggaaatcaca gggtttttgc tgattcaggc ttggcctgaa aacaggacgg
     5701 acctccatgc ctttgagaac ctagaaatca tacgcggcag gaccaagcaa catggtcagt
     5761 tttctcttgc agtcgtcagc ctgaacataa catccttggg attacgctcc ctcaaggaga
     5821 taagtgatgg agatgtgata atttcaggaa acaaaaattt gtgctatgca aatacaataa
     5881 actggaaaaa actgtttggg acctccggtc agaaaaccaa aattataagc aacagaggtg
     5941 aaaacagctg caaggccaca ggccaggtct gccatgcctt gtgctccccc gagggctgct
     6001 ggggcccgga gcccagggac tgcgtctctt gccggaatgt cagccgaggc agggaatgcg
     6061 tggacaagtg caaccttctg gagggtgagc caagggagtt tgtggagaac tctgagtgca
     6121 tacagtgcca cccagagtgc ctgcctcagg ccatgaacat cacctgcaca ggacggggac
     6181 cagacaactg tatccagtgt gcccactaca ttgacggccc ccactgcgtc aagacctgcc
     6241 cggcaggagt catgggagaa aacaacaccc tggtctggaa gtacgcagac gccggccatg
     6301 tgtgccacct gtgccatcca aactgcacct acggatgcac tgggccaggt cttgaaggct
     6361 gtccaacgaa tgggcctaag atcccgtcca tcgccactgg gatggtgggg gccctcctct
     6421 tgctgc
//
