LOCUS       sequence-168844-        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"
     rep_origin      55..643
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     544..563
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     797..814
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(973..1000)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1092..1178)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1227..1249
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1241..1258
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1242..1258
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1274..1373
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(1376..1447,1448..3310,3311..3379,3380..5008)
                     /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"
     protein_bind    complement(5010..5109)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5126..5145)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5127..5145)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5150..5166)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5150..5166)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5597..6388
                     /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 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
       61 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
      121 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
      181 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
      241 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
      301 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
      361 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
      421 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
      481 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
      541 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
      601 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
      661 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
      721 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
      781 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
      841 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
      901 ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc aggaagagtt
      961 tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca
     1021 gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc
     1081 gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga
     1141 aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct
     1201 cgcgttaacg ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct
     1261 taagctcggg ccccaaataa tgattttatt ttgactgata gtgacctgtt cgttgcaaca
     1321 aattgatgag caatgctttt ttataatgcc aactttgtac aaaaaagttg gcaccatgcg
     1381 accctccggg acggccgggg cagcgctcct ggcgctgctg gctgcgctct gcccggcgag
     1441 tcgggctctg gaggaaaaga aagtttgcca aggcacgagt aacaagctca cgcagttggg
     1501 cacttttgaa gatcattttc tcagcctcca gaggatgttc aataactgtg aggtggtcct
     1561 tgggaatttg gaaattacct atgtgcagag gaattatgat ctttccttct taaagaccat
     1621 ccaggaggtg gctggttatg tcctcattgc cctcaacaca gtggagcgaa ttcctttgga
     1681 aaacctgcag atcatcagag gaaatatgta ctacgaaaat tcctatgcct tagcagtctt
     1741 atctaactat gatgcaaata aaaccggact gaaggagctg cccatgagaa atttacagga
     1801 aatcctgcat ggcgccgtgc ggttcagcaa caaccctgcc ctgtgcaacg tggagagcat
     1861 ccagtggcgg gacatagtca gcagtgactt tctcagcaac atgtcgatgg acttccagaa
     1921 ccacctgggc agctgccaaa agtgtgatcc aagctgtccc aatgggagct gctggggtgc
     1981 aggagaggag aactgccaga aactgaccaa aatcatctgt gcccagcagt gctccgggcg
     2041 ctgccgtggc aagtccccca gtgactgctg ccacaaccag tgtgctgcag gctgcacagg
     2101 cccccgggag agcgactgcc tggtctgccg caaattccga gacgaagcca cgtgcaagga
     2161 cacctgcccc ccactcatgc tctacaaccc caccacgtac cagatggatg tgaaccccga
     2221 gggcaaatac agctttggtg ccacctgcgt gaagaagtgt ccccgtaatt atgtggtgac
     2281 agatcacggc tcgtgcgtcc gagcctgtgg ggccgacagc tatgagatgg aggaagacgg
     2341 cgtccgcaag tgtaagaagt gcgaagggcc ttgccgcaaa gtgtgtaacg gaataggtat
     2401 tggtgaattt aaagactcac tctccataaa tgctacgaat attaaacact tcaaaaactg
     2461 cacctccatc agtggcgatc tccacatcct gccggtggca tttaggggtg actccttcac
     2521 acatactcct cctctggatc cacaggaact ggatattctg aaaaccgtaa aggaaatcac
     2581 agggtttttg ctgattcagg cttggcctga aaacaggacg gacctccatg cctttgagaa
     2641 cctagaaatc atacgcggca ggaccaagca acatggtcag ttttctcttg cagtcgtcag
     2701 cctgaacata acatccttgg gattacgctc cctcaaggag ataagtgatg gagatgtgat
     2761 aatttcagga aacaaaaatt tgtgctatgc aaatacaata aactggaaaa aactgtttgg
     2821 gacctccggt cagaaaacca aaattataag caacagaggt gaaaacagct gcaaggccac
     2881 aggccaggtc tgccatgcct tgtgctcccc cgagggctgc tggggcccgg agcccaggga
     2941 ctgcgtctct tgccggaatg tcagccgagg cagggaatgc gtggacaagt gcaaccttct
     3001 ggagggtgag ccaagggagt ttgtggagaa ctctgagtgc atacagtgcc acccagagtg
     3061 cctgcctcag gccatgaaca tcacctgcac aggacgggga ccagacaact gtatccagtg
     3121 tgcccactac attgacggcc cccactgcgt caagacctgc ccggcaggag tcatgggaga
     3181 aaacaacacc ctggtctgga agtacgcaga cgccggccat gtgtgccacc tgtgccatcc
     3241 aaactgcacc tacggatgca ctgggccagg tcttgaaggc tgtccaacga atgggcctaa
     3301 gatcccgtcc atcgccactg ggatggtggg ggccctcctc ttgctgctgg tggtggccct
     3361 ggggatcggc ctcttcatgc gaaggcgcca catcgttcgg aagcgcacgc tgcggaggct
     3421 gctgcaggag agggagcttg tggagcctct tacacccagt ggagaagctc ccaaccaagc
     3481 tctcttgagg atcttgaagg aaactgaatt caaaaagatc aaagtgctgg gctccggtgc
     3541 gttcggcacg gtgtataagg gactctggat cccagaaggt gagaaagtta aaattcccgt
     3601 cgctatcaag gaattaagag aagcaacatc tccgaaagcc aacaaggaaa tcctcgatga
     3661 agcctacgtg atggccagcg tggacaaccc ccacgtgtgc cgcctgctgg gcatctgcct
     3721 cacctccacc gtgcaactca tcacgcagct catgcccttc ggctgcctcc tggactatgt
     3781 ccgggaacac aaagacaata ttggctccca gtacctgctc aactggtgtg tgcagatcgc
     3841 aaagggcatg aactacttgg aggaccgtcg cttggtgcac cgcgacctgg cagccaggaa
     3901 cgtactggtg aaaacaccgc agcatgtcaa gatcacagat tttgggctgg ccaaactgct
     3961 gggtgcggaa gagaaagaat accatgcaga aggaggcaaa gtgcctatca agtggatggc
     4021 attggaatca attttacaca gaatctatac ccaccagagt gatgtctgga gctacggggt
     4081 gaccgtttgg gagttgatga cctttggatc caagccatat gacggaatcc ctgccagcga
     4141 gatctcctcc atcctggaga aaggagaacg cctccctcag ccacccatat gtaccatcga
     4201 tgtctacatg atcatggtca agtgctggat gatagacgca gatagtcgcc caaagttccg
     4261 tgagttgatc atcgaattct ccaaaatggc ccgagacccc cagcgctacc ttgtcattca
     4321 gggggatgaa agaatgcatt tgccaagtcc tacagactcc aacttctacc gtgccctgat
     4381 ggatgaagaa gacatggacg acgtggtgga tgccgacgag tacctcatcc cacagcaggg
     4441 cttcttcagc agcccctcca cgtcacggac tcccctcctg agctctctga gtgcaaccag
     4501 caacaattcc accgtggctt gcattgatag aaatgggctg caaagctgtc ccatcaagga
     4561 agacagcttc ttgcagcgat acagctcaga ccccacaggc gccttgactg aggacagcat
     4621 agacgacacc ttcctcccag tgcctgaata cataaaccag tccgttccca aaaggcccgc
     4681 tggctctgtg cagaatcctg tctatcacaa tcagcctctg aaccccgcgc ccagcagaga
     4741 cccacactac caggaccccc acagcactgc agtgggcaac cccgagtatc tcaacactgt
     4801 ccagcccacc tgtgtcaaca gcacattcga cagccctgcc cactgggccc agaaaggcag
     4861 ccaccaaatt agcctggaca accctgacta ccagcaggac ttctttccca aggaagccaa
     4921 gccaaatggc atctttaagg gctccacagc tgaaaatgca gaatacctaa gggtcgcgcc
     4981 acaaagcagt gaatttattg gagcatgatt gccaactttc ttgtacaaag ttggcattat
     5041 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     5101 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     5161 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     5221 tatatcatca tgcctcctct agaccagcca ggacagaaat gcctcgactt cgctgctacc
     5281 caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc agtggacata
     5341 agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag
     5401 aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga
     5461 ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt
     5521 gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa
     5581 aacaaagtta aacattatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga
     5641 ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg
     5701 ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac
     5761 cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc
     5821 ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga
     5881 catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa
     5941 tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct
     6001 gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga
     6061 tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc
     6121 gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta
     6181 cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg
     6241 cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga
     6301 agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg
     6361 cgagatcacc aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa aatcccttaa
     6421 cgtgag
//
