LOCUS       sequence-202034-        6420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6420)
  AUTHORS   Johannessen CM, Boehm JS, Kim SY, Thomas SR, Wardwell L, Johnson LA,
            Emery CM, Stransky N, Cogdill AP, Barretina J, Caponigro G, 
            Hieronymus H, Murray RR, Salehi-Ashtiani K, Hill DE, Vidal M, Zhao 
            JJ, Yang X, Alkan O, Kim S, Harris JL, Wilson CJ, Myer VE, Finan PM,
            Root DE, Roberts TM, Golub T, Flaherty KT, Dummer R, Weber BL, 
            Sellers WR, Schlegel R, Wargo JA, Hahn WC, Garraway LA
  TITLE     COT drives resistance to RAF inhibition through MAP kinase pathway 
            reactivation.
  JOURNAL   Nature. 2010 Dec 16;468(7326):968-72. doi: 10.1038/nature09627
  PUBMED    21107320
REFERENCE   2  (bases 1 to 6420)
  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"; journalName: 
            "Nature"; date: "2010-12-16- 16"; volume: "468"; issue: "7326"; 
            pages: "968-72"
COMMENT     Sequence Label: pDONR223-EGFR
FEATURES             Location/Qualifiers
     source          1..6420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     17..39
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     31..48
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     32..48
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    64..163
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(166..237,238..2100,2101..2169,2170..3795)
                     /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(3794..3893)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3910..3929)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3911..3929)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3934..3950)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3934..3950)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4381..5172
                     /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      5265..5853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5754..5773
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6007..6024
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6183..6210)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6302..6388)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct taagctcggg
       61 ccccaaataa tgattttatt ttgactgata gtgacctgtt cgttgcaaca aattgatgag
      121 caatgctttt ttataatgcc aactttgtac aaaaaagttg gcaccatgcg accctccggg
      181 acggccgggg cagcgctcct ggcgctgctg gctgcgctct gcccggcgag tcgggctctg
      241 gaggaaaaga aagtttgcca aggcacgagt aacaagctca cgcagttggg cacttttgaa
      301 gatcattttc tcagcctcca gaggatgttc aataactgtg aggtggtcct tgggaatttg
      361 gaaattacct atgtgcagag gaattatgat ctttccttct taaagaccat ccaggaggtg
      421 gctggttatg tcctcattgc cctcaacaca gtggagcgaa ttcctttgga aaacctgcag
      481 atcatcagag gaaatatgta ctacgaaaat tcctatgcct tagcagtctt atctaactat
      541 gatgcaaata aaaccggact gaaggagctg cccatgagaa atttacagga aatcctgcat
      601 ggcgccgtgc ggttcagcaa caaccctgcc ctgtgcaacg tggagagcat ccagtggcgg
      661 gacatagtca gcagtgactt tctcagcaac atgtcgatgg acttccagaa ccacctgggc
      721 agctgccaaa agtgtgatcc aagctgtccc aatgggagct gctggggtgc aggagaggag
      781 aactgccaga aactgaccaa aatcatctgt gcccagcagt gctccgggcg ctgccgtggc
      841 aagtccccca gtgactgctg ccacaaccag tgtgctgcag gctgcacagg cccccgggag
      901 agcgactgcc tggtctgccg caaattccga gacgaagcca cgtgcaagga cacctgcccc
      961 ccactcatgc tctacaaccc caccacgtac cagatggatg tgaaccccga gggcaaatac
     1021 agctttggtg ccacctgcgt gaagaagtgt ccccgtaatt atgtggtgac agatcacggc
     1081 tcgtgcgtcc gagcctgtgg ggccgacagc tatgagatgg aggaagacgg cgtccgcaag
     1141 tgtaagaagt gcgaagggcc ttgccgcaaa gtgtgtaacg gaataggtat tggtgaattt
     1201 aaagactcac tctccataaa tgctacgaat attaaacact tcaaaaactg cacctccatc
     1261 agtggcgatc tccacatcct gccggtggca tttaggggtg actccttcac acatactcct
     1321 cctctggatc cacaggaact ggatattctg aaaaccgtaa aggaaatcac agggtttttg
     1381 ctgattcagg cttggcctga aaacaggacg gacctccatg cctttgagaa cctagaaatc
     1441 atacgcggca ggaccaagca acatggtcag ttttctcttg cagtcgtcag cctgaacata
     1501 acatccttgg gattacgctc cctcaaggag ataagtgatg gagatgtgat aatttcagga
     1561 aacaaaaatt tgtgctatgc aaatacaata aactggaaaa aactgtttgg gacctccggt
     1621 cagaaaacca aaattataag caacagaggt gaaaacagct gcaaggccac aggccaggtc
     1681 tgccatgcct tgtgctcccc cgagggctgc tggggcccgg agcccaggga ctgcgtctct
     1741 tgccggaatg tcagccgagg cagggaatgc gtggacaagt gcaaccttct ggagggtgag
     1801 ccaagggagt ttgtggagaa ctctgagtgc atacagtgcc acccagagtg cctgcctcag
     1861 gccatgaaca tcacctgcac aggacgggga ccagacaact gtatccagtg tgcccactac
     1921 attgacggcc cccactgcgt caagacctgc ccggcaggag tcatgggaga aaacaacacc
     1981 ctggtctgga agtacgcaga cgccggccat gtgtgccacc tgtgccatcc aaactgcacc
     2041 tacggatgca ctgggccagg tcttgaaggc tgtccaacga atgggcctaa gatcccgtcc
     2101 atcgccactg ggatggtggg ggccctcctc ttgctgctgg tggtggccct ggggatcggc
     2161 ctcttcatgc gaaggcgcca catcgttcgg aagcgcacgc tgcggaggct gctgcaggag
     2221 agggagcttg tggagcctct tacacccagt ggagaagctc ccaaccaagc tctcttgagg
     2281 atcttgaagg aaactgaatt caaaaagatc aaagtgctgg gctccggtgc gttcggcacg
     2341 gtgtataagg gactctggat cccagaaggt gagaaagtta aaattcccgt cgctatcaag
     2401 gaattaagag aagcaacatc tccgaaagcc aacaaggaaa tcctcgatga agcctacgtg
     2461 atggccagcg tggacaaccc ccacgtgtgc cgcctgctgg gcatctgcct cacctccacc
     2521 gtgcaactca tcacgcagct catgcccttc ggctgcctcc tggactatgt ccgggaacac
     2581 aaagacaata ttggctccca gtacctgctc aactggtgtg tgcagatcgc aaagggcatg
     2641 aactacttgg aggaccgtcg cttggtgcac cgcgacctgg cagccaggaa cgtactggtg
     2701 aaaacaccgc agcatgtcaa gatcacagat tttgggctgg ccaaactgct gggtgcggaa
     2761 gagaaagaat accatgcaga aggaggcaaa gtgcctatca agtggatggc attggaatca
     2821 attttacaca gaatctatac ccaccagagt gatgtctgga gctacggggt gaccgtttgg
     2881 gagttgatga cctttggatc caagccatat gacggaatcc ctgccagcga gatctcctcc
     2941 atcctggaga aaggagaacg cctccctcag ccacccatat gtaccatcga tgtctacatg
     3001 atcatggtca agtgctggat gatagacgca gatagtcgcc caaagttccg tgagttgatc
     3061 atcgaattct ccaaaatggc ccgagacccc cagcgctacc ttgtcattca gggggatgaa
     3121 agaatgcatt tgccaagtcc tacagactcc aacttctacc gtgccctgat ggatgaagaa
     3181 gacatggacg acgtggtgga tgccgacgag tacctcatcc cacagcaggg cttcttcagc
     3241 agcccctcca cgtcacggac tcccctcctg agctctctga gtgcaaccag caacaattcc
     3301 accgtggctt gcattgatag aaatgggctg caaagctgtc ccatcaagga agacagcttc
     3361 ttgcagcgat acagctcaga ccccacaggc gccttgactg aggacagcat agacgacacc
     3421 ttcctcccag tgcctgaata cataaaccag tccgttccca aaaggcccgc tggctctgtg
     3481 cagaatcctg tctatcacaa tcagcctctg aaccccgcgc ccagcagaga cccacactac
     3541 caggaccccc acagcactgc agtgggcaac cccgagtatc tcaacactgt ccagcccacc
     3601 tgtgtcaaca gcacattcga cagccctgcc cactgggccc agaaaggcag ccaccaaatt
     3661 agcctggaca accctgacta ccagcaggac ttctttccca aggaagccaa gccaaatggc
     3721 atctttaagg gctccacagc tgaaaatgca gaatacctaa gggtcgcgcc acaaagcagt
     3781 gaatttattg gagcaccaac tttcttgtac aaagttggca ttataagaaa gcattgctta
     3841 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca
     3901 gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg
     3961 cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgcctc
     4021 ctctagacca gccaggacag aaatgcctcg acttcgctgc tacccaaggt tgccgggtga
     4081 cgcacaccgt ggaaacggat gaaggcacga acccagtgga cataagcctg ttcggttcgt
     4141 aagctgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt gaccgaacgc
     4201 agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttggggta
     4261 cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg
     4321 ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatt
     4381 atgagggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt tggcgtcatc
     4441 gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc agtggatggc
     4501 ggcctgaagc cacacagtga tattgatttg ctggttacgg tgaccgtaag gcttgatgaa
     4561 acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc tggagagagc
     4621 gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat tccgtggcgt
     4681 tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat tcttgcaggt
     4741 atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa agcaagagaa
     4801 catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt tcctgaacag
     4861 gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc cgactgggct
     4921 ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc agtaaccggc
     4981 aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat
     5041 cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga tcgcttggcc
     5101 tcgcgcgcag atcagttgga agaatttgtc cactacgtga aaggcgagat caccaaggta
     5161 gtcggcaaat aaccctcgag ccacccatga ccaaaatccc ttaacgtgag ttacgcgtcg
     5221 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5281 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5341 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5401 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5461 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5521 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5581 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5641 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5701 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5761 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5821 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5881 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5941 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6001 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6061 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     6121 ggcagtgagc gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg
     6181 caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg
     6241 ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg
     6301 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
     6361 cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg
//
