LOCUS       Exported                6420 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Gateway Donor vector containing EGFR, part of the Target Accelerator
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    pDONR223_EGFR_p.T790M_p.L858R
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6420)
  AUTHORS   Berger AH, Brooks AN, Wu X, Shrestha Y, Chouinard C, Piccioni F, 
            Bagul M, Kamburov A, Imielinski M, Hogstrom L, Zhu C, Yang X, Pantel
            S, Sakai R, Watson J, Kaplan N, Campbell JD, Singh S, Root DE, 
            Narayan R, Natoli T, Lahr DL, Tirosh I, Tamayo P, Getz G, Wong B, 
            Doench J, Subramanian A, Golub TR, Meyerson M, Boehm JS
  TITLE     High-throughput Phenotyping of Lung Cancer Somatic Mutations.
  JOURNAL   Cancer Cell. 2016 Aug 8;30(2):214-28. doi: 
            10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28.
  PUBMED    27478040
REFERENCE   2  (bases 1 to 6420)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6420
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     43..65
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     57..74
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     58..74
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    90..189
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             189..3818
                     /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
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLIMQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGRAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     protein_bind    complement(3820..3919)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3936..3955)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3937..3955)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3960..3976)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3960..3976)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4407..5198
                     /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      5291..5879
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5780..5799
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6033..6050
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      6209..6236
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      6328..6414
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
       61 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      121 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
      181 aagttggcat gcgaccctcc gggacggccg gggcagcgct cctggcgctg ctggctgcgc
      241 tctgcccggc gagtcgggct ctggaggaaa agaaagtttg ccaaggcacg agtaacaagc
      301 tcacgcagtt gggcactttt gaagatcatt ttctcagcct ccagaggatg ttcaataact
      361 gtgaggtggt ccttgggaat ttggaaatta cctatgtgca gaggaattat gatctttcct
      421 tcttaaagac catccaggag gtggctggtt atgtcctcat tgccctcaac acagtggagc
      481 gaattccttt ggaaaacctg cagatcatca gaggaaatat gtactacgaa aattcctatg
      541 ccttagcagt cttatctaac tatgatgcaa ataaaaccgg actgaaggag ctgcccatga
      601 gaaatttaca ggaaatcctg catggcgccg tgcggttcag caacaaccct gccctgtgca
      661 acgtggagag catccagtgg cgggacatag tcagcagtga ctttctcagc aacatgtcga
      721 tggacttcca gaaccacctg ggcagctgcc aaaagtgtga tccaagctgt cccaatggga
      781 gctgctgggg tgcaggagag gagaactgcc agaaactgac caaaatcatc tgtgcccagc
      841 agtgctccgg gcgctgccgt ggcaagtccc ccagtgactg ctgccacaac cagtgtgctg
      901 caggctgcac aggcccccgg gagagcgact gcctggtctg ccgcaaattc cgagacgaag
      961 ccacgtgcaa ggacacctgc cccccactca tgctctacaa ccccaccacg taccagatgg
     1021 atgtgaaccc cgagggcaaa tacagctttg gtgccacctg cgtgaagaag tgtccccgta
     1081 attatgtggt gacagatcac ggctcgtgcg tccgagcctg tggggccgac agctatgaga
     1141 tggaggaaga cggcgtccgc aagtgtaaga agtgcgaagg gccttgccgc aaagtgtgta
     1201 acggaatagg tattggtgaa tttaaagact cactctccat aaatgctacg aatattaaac
     1261 acttcaaaaa ctgcacctcc atcagtggcg atctccacat cctgccggtg gcatttaggg
     1321 gtgactcctt cacacatact cctcctctgg atccacagga actggatatt ctgaaaaccg
     1381 taaaggaaat cacagggttt ttgctgattc aggcttggcc tgaaaacagg acggacctcc
     1441 atgcctttga gaacctagaa atcatacgcg gcaggaccaa gcaacatggt cagttttctc
     1501 ttgcagtcgt cagcctgaac ataacatcct tgggattacg ctccctcaag gagataagtg
     1561 atggagatgt gataatttca ggaaacaaaa atttgtgcta tgcaaataca ataaactgga
     1621 aaaaactgtt tgggacctcc ggtcagaaaa ccaaaattat aagcaacaga ggtgaaaaca
     1681 gctgcaaggc cacaggccag gtctgccatg ccttgtgctc ccccgagggc tgctggggcc
     1741 cggagcccag ggactgcgtc tcttgccgga atgtcagccg aggcagggaa tgcgtggaca
     1801 agtgcaacct tctggagggt gagccaaggg agtttgtgga gaactctgag tgcatacagt
     1861 gccacccaga gtgcctgcct caggccatga acatcacctg cacaggacgg ggaccagaca
     1921 actgtatcca gtgtgcccac tacattgacg gcccccactg cgtcaagacc tgcccggcag
     1981 gagtcatggg agaaaacaac accctggtct ggaagtacgc agacgccggc catgtgtgcc
     2041 acctgtgcca tccaaactgc acctacggat gcactgggcc aggtcttgaa ggctgtccaa
     2101 cgaatgggcc taagatcccg tccatcgcca ctgggatggt gggggccctc ctcttgctgc
     2161 tggtggtggc cctggggatc ggcctcttca tgcgaaggcg ccacatcgtt cggaagcgca
     2221 cgctgcggag gctgctgcag gagagggagc ttgtggagcc tcttacaccc agtggagaag
     2281 ctcccaacca agctctcttg aggatcttga aggaaactga attcaaaaag atcaaagtgc
     2341 tgggctccgg tgcgttcggc acggtgtata agggactctg gatcccagaa ggtgagaaag
     2401 ttaaaattcc cgtcgctatc aaggaattaa gagaagcaac atctccgaaa gccaacaagg
     2461 aaatcctcga tgaagcctac gtgatggcca gcgtggacaa cccccacgtg tgccgcctgc
     2521 tgggcatctg cctcacctcc accgtgcaac tcatcatgca gctcatgccc ttcggctgcc
     2581 tcctggacta tgtccgggaa cacaaagaca atattggctc ccagtacctg ctcaactggt
     2641 gtgtgcagat cgcaaagggc atgaactact tggaggaccg tcgcttggtg caccgcgacc
     2701 tggcagccag gaacgtactg gtgaaaacac cgcagcatgt caagatcaca gattttggga
     2761 gggccaaact gctgggtgcg gaagagaaag aataccatgc agaaggaggc aaagtgccta
     2821 tcaagtggat ggcattggaa tcaattttac acagaatcta tacccaccag agtgatgtct
     2881 ggagctacgg ggtgaccgtt tgggagttga tgacctttgg atccaagcca tatgacggaa
     2941 tccctgccag cgagatctcc tccatcctgg agaaaggaga acgcctccct cagccaccca
     3001 tatgtaccat cgatgtctac atgatcatgg tcaagtgctg gatgatagac gcagatagtc
     3061 gcccaaagtt ccgtgagttg atcatcgaat tctccaaaat ggcccgagac ccccagcgct
     3121 accttgtcat tcagggggat gaaagaatgc atttgccaag tcctacagac tccaacttct
     3181 accgtgccct gatggatgaa gaagacatgg acgacgtggt ggatgccgac gagtacctca
     3241 tcccacagca gggcttcttc agcagcccct ccacgtcacg gactcccctc ctgagctctc
     3301 tgagtgcaac cagcaacaat tccaccgtgg cttgcattga tagaaatggg ctgcaaagct
     3361 gtcccatcaa ggaagacagc ttcttgcagc gatacagctc agaccccaca ggcgccttga
     3421 ctgaggacag catagacgac accttcctcc cagtgcctga atacataaac cagtccgttc
     3481 ccaaaaggcc cgctggctct gtgcagaatc ctgtctatca caatcagcct ctgaaccccg
     3541 cgcccagcag agacccacac taccaggacc cccacagcac tgcagtgggc aaccccgagt
     3601 atctcaacac tgtccagccc acctgtgtca acagcacatt cgacagccct gcccactggg
     3661 cccagaaagg cagccaccaa attagcctgg acaaccctga ctaccagcag gacttctttc
     3721 ccaaggaagc caagccaaat ggcatcttta agggctccac agctgaaaat gcagaatacc
     3781 taagggtcgc gccacaaagc agtgaattta ttggagcatg cccaactttc ttgtacaaag
     3841 ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     3901 aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg
     3961 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     4021 aagataaaaa tatatcatca tgcctcctct agaccagcca ggacagaaat gcctcgactt
     4081 cgctgctacc caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc
     4141 agtggacata agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca
     4201 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     4261 cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc
     4321 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca
     4381 gtcgccctaa aacaaagtta aacattatga gggaagcggt gatcgccgaa gtatcgactc
     4441 aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac
     4501 atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg
     4561 ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg
     4621 aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg
     4681 tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat
     4741 ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg
     4801 ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg
     4861 aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc
     4921 tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc
     4981 gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg
     5041 caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc
     5101 ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact
     5161 acgtgaaagg cgagatcacc aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa
     5221 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     5281 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5341 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5401 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5461 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5521 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5581 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5641 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5701 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5761 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5821 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5881 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5941 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     6001 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     6061 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     6121 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
     6181 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
     6241 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
     6301 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
     6361 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
//
