LOCUS       sequence-168660-        6438 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 6438)
  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 6438)
  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.G719D
FEATURES             Location/Qualifiers
     source          1..6438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      21..609
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     510..529
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     763..780
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(939..966)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1058..1144)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1193..1215
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1207..1224
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1208..1224
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1240..1339
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(1342..1413,1414..3276,3277..3345,3346..4971)
                     /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
                     SGEAPNQALLRILKETEFKKIKVLDSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(4988..5087)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5104..5123)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5105..5123)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5128..5144)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5128..5144)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5575..6366
                     /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"
ORIGIN
        1 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
       61 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
      121 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
      181 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
      241 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
      301 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
      361 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
      421 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
      481 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
      541 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
      601 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
      661 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
      721 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
      781 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
      841 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
      901 acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg
      961 ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg
     1021 gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt
     1081 tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta
     1141 tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca
     1201 cgacgttgta aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga
     1261 ctgatagtga cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact
     1321 ttgtacaaaa aagttggcac catgcgaccc tccgggacgg ccggggcagc gctcctggcg
     1381 ctgctggctg cgctctgccc ggcgagtcgg gctctggagg aaaagaaagt ttgccaaggc
     1441 acgagtaaca agctcacgca gttgggcact tttgaagatc attttctcag cctccagagg
     1501 atgttcaata actgtgaggt ggtccttggg aatttggaaa ttacctatgt gcagaggaat
     1561 tatgatcttt ccttcttaaa gaccatccag gaggtggctg gttatgtcct cattgccctc
     1621 aacacagtgg agcgaattcc tttggaaaac ctgcagatca tcagaggaaa tatgtactac
     1681 gaaaattcct atgccttagc agtcttatct aactatgatg caaataaaac cggactgaag
     1741 gagctgccca tgagaaattt acaggaaatc ctgcatggcg ccgtgcggtt cagcaacaac
     1801 cctgccctgt gcaacgtgga gagcatccag tggcgggaca tagtcagcag tgactttctc
     1861 agcaacatgt cgatggactt ccagaaccac ctgggcagct gccaaaagtg tgatccaagc
     1921 tgtcccaatg ggagctgctg gggtgcagga gaggagaact gccagaaact gaccaaaatc
     1981 atctgtgccc agcagtgctc cgggcgctgc cgtggcaagt cccccagtga ctgctgccac
     2041 aaccagtgtg ctgcaggctg cacaggcccc cgggagagcg actgcctggt ctgccgcaaa
     2101 ttccgagacg aagccacgtg caaggacacc tgccccccac tcatgctcta caaccccacc
     2161 acgtaccaga tggatgtgaa ccccgagggc aaatacagct ttggtgccac ctgcgtgaag
     2221 aagtgtcccc gtaattatgt ggtgacagat cacggctcgt gcgtccgagc ctgtggggcc
     2281 gacagctatg agatggagga agacggcgtc cgcaagtgta agaagtgcga agggccttgc
     2341 cgcaaagtgt gtaacggaat aggtattggt gaatttaaag actcactctc cataaatgct
     2401 acgaatatta aacacttcaa aaactgcacc tccatcagtg gcgatctcca catcctgccg
     2461 gtggcattta ggggtgactc cttcacacat actcctcctc tggatccaca ggaactggat
     2521 attctgaaaa ccgtaaagga aatcacaggg tttttgctga ttcaggcttg gcctgaaaac
     2581 aggacggacc tccatgcctt tgagaaccta gaaatcatac gcggcaggac caagcaacat
     2641 ggtcagtttt ctcttgcagt cgtcagcctg aacataacat ccttgggatt acgctccctc
     2701 aaggagataa gtgatggaga tgtgataatt tcaggaaaca aaaatttgtg ctatgcaaat
     2761 acaataaact ggaaaaaact gtttgggacc tccggtcaga aaaccaaaat tataagcaac
     2821 agaggtgaaa acagctgcaa ggccacaggc caggtctgcc atgccttgtg ctcccccgag
     2881 ggctgctggg gcccggagcc cagggactgc gtctcttgcc ggaatgtcag ccgaggcagg
     2941 gaatgcgtgg acaagtgcaa ccttctggag ggtgagccaa gggagtttgt ggagaactct
     3001 gagtgcatac agtgccaccc agagtgcctg cctcaggcca tgaacatcac ctgcacagga
     3061 cggggaccag acaactgtat ccagtgtgcc cactacattg acggccccca ctgcgtcaag
     3121 acctgcccgg caggagtcat gggagaaaac aacaccctgg tctggaagta cgcagacgcc
     3181 ggccatgtgt gccacctgtg ccatccaaac tgcacctacg gatgcactgg gccaggtctt
     3241 gaaggctgtc caacgaatgg gcctaagatc ccgtccatcg ccactgggat ggtgggggcc
     3301 ctcctcttgc tgctggtggt ggccctgggg atcggcctct tcatgcgaag gcgccacatc
     3361 gttcggaagc gcacgctgcg gaggctgctg caggagaggg agcttgtgga gcctcttaca
     3421 cccagtggag aagctcccaa ccaagctctc ttgaggatct tgaaggaaac tgaattcaaa
     3481 aagatcaaag tgctggactc cggtgcgttc ggcacggtgt ataagggact ctggatccca
     3541 gaaggtgaga aagttaaaat tcccgtcgct atcaaggaat taagagaagc aacatctccg
     3601 aaagccaaca aggaaatcct cgatgaagcc tacgtgatgg ccagcgtgga caacccccac
     3661 gtgtgccgcc tgctgggcat ctgcctcacc tccaccgtgc aactcatcac gcagctcatg
     3721 cccttcggct gcctcctgga ctatgtccgg gaacacaaag acaatattgg ctcccagtac
     3781 ctgctcaact ggtgtgtgca gatcgcaaag ggcatgaact acttggagga ccgtcgcttg
     3841 gtgcaccgcg acctggcagc caggaacgta ctggtgaaaa caccgcagca tgtcaagatc
     3901 acagattttg ggctggccaa actgctgggt gcggaagaga aagaatacca tgcagaagga
     3961 ggcaaagtgc ctatcaagtg gatggcattg gaatcaattt tacacagaat ctatacccac
     4021 cagagtgatg tctggagcta cggggtgacc gtttgggagt tgatgacctt tggatccaag
     4081 ccatatgacg gaatccctgc cagcgagatc tcctccatcc tggagaaagg agaacgcctc
     4141 cctcagccac ccatatgtac catcgatgtc tacatgatca tggtcaagtg ctggatgata
     4201 gacgcagata gtcgcccaaa gttccgtgag ttgatcatcg aattctccaa aatggcccga
     4261 gacccccagc gctaccttgt cattcagggg gatgaaagaa tgcatttgcc aagtcctaca
     4321 gactccaact tctaccgtgc cctgatggat gaagaagaca tggacgacgt ggtggatgcc
     4381 gacgagtacc tcatcccaca gcagggcttc ttcagcagcc cctccacgtc acggactccc
     4441 ctcctgagct ctctgagtgc aaccagcaac aattccaccg tggcttgcat tgatagaaat
     4501 gggctgcaaa gctgtcccat caaggaagac agcttcttgc agcgatacag ctcagacccc
     4561 acaggcgcct tgactgagga cagcatagac gacaccttcc tcccagtgcc tgaatacata
     4621 aaccagtccg ttcccaaaag gcccgctggc tctgtgcaga atcctgtcta tcacaatcag
     4681 cctctgaacc ccgcgcccag cagagaccca cactaccagg acccccacag cactgcagtg
     4741 ggcaaccccg agtatctcaa cactgtccag cccacctgtg tcaacagcac attcgacagc
     4801 cctgcccact gggcccagaa aggcagccac caaattagcc tggacaaccc tgactaccag
     4861 caggacttct ttcccaagga agccaagcca aatggcatct ttaagggctc cacagctgaa
     4921 aatgcagaat acctaagggt cgcgccacaa agcagtgaat ttattggagc aaagggtggg
     4981 cgcgccgacc cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt
     5041 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat
     5101 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
     5161 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg cctcctctag
     5221 accagccagg acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca
     5281 ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg
     5341 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt
     5401 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct
     5461 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
     5521 agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg
     5581 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc
     5641 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg
     5701 aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg
     5761 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt
     5821 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca
     5881 gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc
     5941 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc
     6001 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta
     6061 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat
     6121 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc
     6181 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc
     6241 gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc
     6301 gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc
     6361 aaataaccct cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
     6421 tgagcgtcag accccgta
//
