LOCUS       sequence-473525-        7650 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 7650)
  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 7650)
  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-ALK
FEATURES             Location/Qualifiers
     source          1..7650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(2460..2559)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2575..2591)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2584..2606)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      2655..2741
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2833..2860
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3019..3036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3190..3778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3270..3289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3871..4662)
                     /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"
     primer_bind     5093..5109
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5093..5109
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5114..5133
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5114..5132
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5150..5249
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tcctccgcct cctgcccact catgcacgct tctgttcaca cggatttctt cttctatcac
       61 attgttctct ccaatgcaga ctttctggat taactggttt gtactggggc aggcgtcctc
      121 tccctgctgc ccaaccagga tgtacagcat gtcatccttc tccaggttga agatgcccag
      181 cacagacacg ccgtgggacc gcatcatggt gttcttcccg cctttcccgc cagcagctcc
      241 gtagcccgag atgctgtagg tgtcggtggc tggcaccttc cagatctgga tgcctttcag
      301 ggggccctcg ctccccacct ccacgctcag gttggagttc tggtaggcgt tgttgcactg
      361 tgcctgggtg gggccatggg gcccgctggc cccacatgtg gtgaacagcc aatgaactgt
      421 agggtcaaag atgggggtct gtcttggtga attttccccg ggtttcagct ccttgtttgg
      481 gtttctctca aacaggtttc ttgatttggg tgctgtattc tgcaggatct tgtcctctcc
      541 gctaatggtg aggtagcagt ccaggctgat ggagatattg tcaaaagcca cgatggctct
      601 ggatccttgt ccccaccatg cgaccatctg cagccagaac ctgtcagaca catcgaggag
      661 aggcaacacc atccactgcc acaggctcaa gccttcatag gcggcgacat gccagaccat
      721 cctgccttgc tccttcccgg ttttgttctc cactagcacc aaggacacgt ttcccctcaa
      781 gactccacga atgagccagg acattcggag ctcacatgga gagctcttga tcggtgcagg
      841 aaacgtagca ctggtcactg tagcactttc agaagcgggg acatcagtgg tactgagcaa
      901 tagagcatgg tcttggtggt cctggaaccg ggcatccttt agggtcctga cctgccactg
      961 aggagtgtgg ggtgacagtg tgccttgggt ccagccacag aagccatctt caaagttgca
     1021 gtaaaaaccc acaggcagtt tccggcacat ctggctctca tcttctccct gggcacagtc
     1081 ctggtggaag tcacaggcct gcccaagctg gaggactgtc ccattccaac aagtgaagga
     1141 gctctgcagg gccatcttgg agcctgggga tgttccttca ctgcagttct tcagggcaaa
     1201 gaagtccact gcagacaagc tgcggtttcc actggagatg tattccaggg ccactcgaaa
     1261 tgggttgtct ggacgcccga ttcttccctg gagcactgtc caaccatgct tccctggagt
     1321 gggcatcagg aggatctctc ttgcagcctc gttgtggggc agcagctggg caatgtacct
     1381 tccagagggc tgcaggtgcc tgtgcaccga gacggccagt gtgcagtgct cactgctgct
     1441 cctcatccac ggactcagga tggtgtgctt ggagtcagct gaggtgttga gaaggagaaa
     1501 ggagcctctg ggcatctcct tagaacgctc tgccccaggc ccatccagca agtccatctg
     1561 ggaggcctcc tcggagggga tgcggcgcca ggaccagctc tggttcctga ggtcatgcag
     1621 tggaggggaa tactccagct cacaggggaa gtcaaagctg cactccagac catatcggct
     1681 gcgatgagac aggaaaggga aggagtcttt cattatccag gtgagattcc atgtaaaata
     1741 atcaggagaa ggagatggca tgtttgttgg tgattccaag gagctatgac cagtcccgaa
     1801 gatctggaag agaaggcggg gctgggaggc gcgaattgcc gcggacagcc ttccctctct
     1861 gcccacttcc gacgccttct tctcgggcat caggcggatc ctcagtcgcc cttcgccttg
     1921 gcgaatccac caactgaaca gctcgctgag attgaactgg agcagcccca cagccgcctc
     1981 cccggggggc ccgacgcaac cctccaagat cgcctcctcg cccagctcca gcaccaactg
     2041 cttggcacgc cggagcttgc gcacggagcc gcccttcagc accctggaca gcgtccgggc
     2101 ctctgccggg gctggtgaac cggcggtcca ggagaccccc ggcgccggcc ccagcaacct
     2161 gagcagcggg gcgcagtcca gagctagcga gccgcgggcc tcgggcctgc cagccttcag
     2221 ctccgaggag gatggtggca gcagtaggtc ccgggcgtag acacggaaga gcgagggcac
     2281 cacgaagtca actgccagac tcttcctctg caggcgcgag tagctgagtg gctcccgggg
     2341 ctgcagcggc gaccccgcag ctggggagcc cgcgcgctgg ccggtcccca tcccggagcc
     2401 cacagctgcc gtggaaagca gcagcggcag cagccacagg agcccgatgg ctcccatggt
     2461 gccaactttt ttgtacaaag ttggcattat aaaaaagcat tgctcatcaa tttgttgcaa
     2521 cgaacaggtc actatcagtc aaaataaaat cattatttgg ggcccgagct taagactggc
     2581 cgtcgtttta caacgtcgtg actgggaaaa catccatgct agcgttaacg cgagagtagg
     2641 gaactgccag gcatcaaata aaacgaaagg ctcagtcgga agactgggcc tttcgtttta
     2701 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     2761 acgttgtgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     2821 atcaaactaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttc
     2881 ctggctagcg gtacgcgtat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2941 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3001 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3061 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3121 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3181 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3241 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3301 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3361 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3421 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3481 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3541 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3601 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3661 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3721 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3781 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgacgcgt aactcacgtt
     3841 aagggatttt ggtcatgggt ggctcgaggg ttatttgccg actaccttgg tgatctcgcc
     3901 tttcacgtag tggacaaatt cttccaactg atctgcgcgc gaggccaagc gatcttcttc
     3961 ttgtccaaga taagcctgtc tagcttcaag tatgacgggc tgatactggg ccggcaggcg
     4021 ctccattgcc cagtcggcag cgacatcctt cggcgcgatt ttgccggtta ctgcgctgta
     4081 ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg ccagcccagt cgggcggcga
     4141 gttccatagc gttaaggttt catttagcgc ctcaaataga tcctgttcag gaaccggatc
     4201 aaagagttcc tccgccgctg gacctaccaa ggcaacgcta tgttctcttg cttttgtcag
     4261 caagatagcc agatcaatgt cgatcgtggc tggctcgaag atacctgcaa gaatgtcatt
     4321 gcgctgccat tctccaaatt gcagttcgcg cttagctgga taacgccacg gaatgatgtc
     4381 gtcgtgcaca acaatggtga cttctacagc gcggagaatc tcgctctctc caggggaagc
     4441 cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa gccttacggt
     4501 caccgtaacc agcaaatcaa tatcactgtg tggcttcagg ccgccatcca ctgcggagcc
     4561 gtacaaatgt acggccagca acgtcggttc gagatggcgc tcgatgacgc caactacctc
     4621 tgatagttga gtcgatactt cggcgatcac cgcttccctc ataatgttta actttgtttt
     4681 agggcgactg ccctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac
     4741 ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtaccccaa aaaaacagtc
     4801 ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct
     4861 ggaccagttg cgtgagcgca tacgctactt gcattacagc ttacgaaccg aacaggctta
     4921 tgtccactgg gttcgtgcct tcatccgttt ccacggtgtg cgtcacccgg caaccttggg
     4981 tagcagcgaa gtcgaggcat ttctgtcctg gctggtctag aggaggcatg atgatatatt
     5041 tttatcttgt gcaatgtaac atcagagatt ttgagacacg ggccagagct gccaggaaac
     5101 agctatgacc atgtaatacg actcactata ggggatatca gctggatggc aaataatgat
     5161 tttattttga ctgatagtga cctgttcgtt gcaacaaatt gataagcaat gctttcttat
     5221 aatgccaact ttgtacaaga aagttggggg cccaggctgg ttcatgctat tcttgctttt
     5281 cagaatggta tcctcgtaat gaccagctcc aggggcagta gcggcttcta agggcaagcc
     5341 ctgttgctgg tagccgtaat tgacattccc acaagggaag tgacgtagcc tgaacagagg
     5401 tacctccttc atattggcag tcagcgaaga gggctctagg agcagtgagg cccccggaag
     5461 tctcccagtt gcaacgttag gtgggacagt acagcttccc tccagcccca ggttacccct
     5521 gtcgtgtggc tccttctttg ctataggatt attctttttg gtgggtttct ctgtaaacca
     5581 ggagccgtac gttgggttcc acaagctggt gggcttgttt ctggatccgt ggaccttgtg
     5641 caactccgaa ggagggttgg actgagagaa tgccatattc acgtgtcccc cttccacggc
     5701 cggccctcta gggactcgaa cagagacctc tgcagctgtg ggtttctttg cagccttgcc
     5761 agaggaggtg gtaggcagag gtggtggggc agctgggctg cgctcctcct cccgttttgc
     5821 ctgttgagag accaggagag gaggaacccc ctcagggtcc ttgggcctca caggcacttt
     5881 ctcttcctct tccacaagtg gaccatattc tatcggcaaa gcggtgttga ttacatccgg
     5941 gtcctgggtg cagtattcaa tcctctccaa aatgatggca aagttgggcc tgtcttcagg
     6001 ctgatgttgc cagcactgag tcattatccg gtatacaggc ccagggcagt tcttgggtgg
     6061 gtccatccgg cctccactgg tgacaaactc cagaacttcc tggttgcttt tgctggggta
     6121 tggcatatat ccaagagaaa agatttccca tagcagcact ccaaaggacc atgtgtctgt
     6181 tttagaagtg aatattcctt ccatgaaggc ctctgggggc atccacttaa ctggcagcat
     6241 ggcacagcct ccctttctat agtagctcgc cctgtagatg tctcgggcca tcccgaagtc
     6301 tccaatcttg gccactcttc cagggcctgg acaggtcaag aggcagtttc tggcagcaat
     6361 gtctcggtgg atgaagtggt tttcctccaa atactgacag ccacaggcaa tgtcccgagc
     6421 cacgtgcaga aggtccagca tggccaggga ggagggctgg ctcgggcgag ggcgggtctc
     6481 tcggaggaag gacttgaggt ctccccccgc catgagctcc agcaggatga accggggcag
     6541 ggattgcagg ctcaccccaa tgcagcgaac aatgttctgg tggttgaatt tgctgatgat
     6601 cagggcttcc atgaggaaat ccagttcgtc ctgttcagag cacacttcag gcagcgtctt
     6661 cacagccact tgcagggggc ttgggtcgtt gggcattccg gacacctggc cttcatacac
     6721 ctccccaaag gcgccatggc ccagaccccg aatgagggtg atgtttttcc gcggcacctc
     6781 cttcaggtca ctgatggagg aggtcttgcc agcaaagcag tagttggggt tgtagtcggt
     6841 catgatggtc gaggtgcgga gcttgctcag cttgtactca gggctctgca gctccatctg
     6901 catggcttgc agctcctggt gcttccggcg gtacacaatc atgatgccgg agaaagccag
     6961 gaccagggcg gccacgaggg cagaggtcac cacagagagg atcagcgaga gtggcaggtg
     7021 tggctccggg gtgggtgaca caatgcagga gacgccatcc tcagccagca ccgtcccgtg
     7081 gtcacagaag cagatgacct tgtggctttc agggtccatg tgacattcgt ctacctcaca
     7141 gtgactgcag tttagataat gcttaatatt cacttccccg tggccttcca tcacttttaa
     7201 agctggggtg tacaggatgc ccagtggact gatgaaggaa accccatctt ccccatccat
     7261 ttcggggtca ttgtttgagg ctgcattgcc gcctatatat cctccgcctc ctccacctga
     7321 ggagcacccc cctccacccc ctccgaaacc ccctcttgtc tcccaccccc acttcttcat
     7381 ggcctggggg caggaatgtc ctccggtggc accctcctgc aaagattttc cggcccagag
     7441 caaggaagtg ttatcattcc agccacctcc accacctgcg gctccggaat tgccgtttag
     7501 ccctagaacc gaggagttat tctccagtct ctctgggtgg aacgtgtctg tcttggcccc
     7561 gtaggccctg ccgccacctc cggctgcaat gatcaggggc accggcactc catccttcat
     7621 cttaaatacg taggtggctc caccccctcc
//
