LOCUS       sequence-140188-        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"
     terminator      complement(149..176)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(268..354)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     403..425
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     418..434
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    450..549
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(5410..5509)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5526..5545)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5527..5545)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5550..5566)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5550..5566)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5997..6788
                     /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      6881..7469
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7370..7389
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7623..7640
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
      121 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
      181 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
      241 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
      301 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
      361 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
      421 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      481 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
      541 aagttggcac catgggagcc atcgggctcc tgtggctgct gccgctgctg ctttccacgg
      601 cagctgtggg ctccgggatg gggaccggcc agcgcgcggg ctccccagct gcggggtcgc
      661 cgctgcagcc ccgggagcca ctcagctact cgcgcctgca gaggaagagt ctggcagttg
      721 acttcgtggt gccctcgctc ttccgtgtct acgcccggga cctactgctg ccaccatcct
      781 cctcggagct gaaggctggc aggcccgagg cccgcggctc gctagctctg gactgcgccc
      841 cgctgctcag gttgctgggg ccggcgccgg gggtctcctg gaccgccggt tcaccagccc
      901 cggcagaggc ccggacgctg tccagggtgc tgaagggcgg ctccgtgcgc aagctccggc
      961 gtgccaagca gttggtgctg gagctgggcg aggaggcgat cttggagggt tgcgtcgggc
     1021 cccccgggga ggcggctgtg gggctgctcc agttcaatct cagcgagctg ttcagttggt
     1081 ggattcgcca aggcgaaggg cgactgagga tccgcctgat gcccgagaag aaggcgtcgg
     1141 aagtgggcag agagggaagg ctgtccgcgg caattcgcgc ctcccagccc cgccttctct
     1201 tccagatctt cgggactggt catagctcct tggaatcacc aacaaacatg ccatctcctt
     1261 ctcctgatta ttttacatgg aatctcacct ggataatgaa agactccttc cctttcctgt
     1321 ctcatcgcag ccgatatggt ctggagtgca gctttgactt cccctgtgag ctggagtatt
     1381 cccctccact gcatgacctc aggaaccaga gctggtcctg gcgccgcatc ccctccgagg
     1441 aggcctccca gatggacttg ctggatgggc ctggggcaga gcgttctaag gagatgccca
     1501 gaggctcctt tctccttctc aacacctcag ctgactccaa gcacaccatc ctgagtccgt
     1561 ggatgaggag cagcagtgag cactgcacac tggccgtctc ggtgcacagg cacctgcagc
     1621 cctctggaag gtacattgcc cagctgctgc cccacaacga ggctgcaaga gagatcctcc
     1681 tgatgcccac tccagggaag catggttgga cagtgctcca gggaagaatc gggcgtccag
     1741 acaacccatt tcgagtggcc ctggaataca tctccagtgg aaaccgcagc ttgtctgcag
     1801 tggacttctt tgccctgaag aactgcagtg aaggaacatc cccaggctcc aagatggccc
     1861 tgcagagctc cttcacttgt tggaatggga cagtcctcca gcttgggcag gcctgtgact
     1921 tccaccagga ctgtgcccag ggagaagatg agagccagat gtgccggaaa ctgcctgtgg
     1981 gtttttactg caactttgaa gatggcttct gtggctggac ccaaggcaca ctgtcacccc
     2041 acactcctca gtggcaggtc aggaccctaa aggatgcccg gttccaggac caccaagacc
     2101 atgctctatt gctcagtacc actgatgtcc ccgcttctga aagtgctaca gtgaccagtg
     2161 ctacgtttcc tgcaccgatc aagagctctc catgtgagct ccgaatgtcc tggctcattc
     2221 gtggagtctt gaggggaaac gtgtccttgg tgctagtgga gaacaaaacc gggaaggagc
     2281 aaggcaggat ggtctggcat gtcgccgcct atgaaggctt gagcctgtgg cagtggatgg
     2341 tgttgcctct cctcgatgtg tctgacaggt tctggctgca gatggtcgca tggtggggac
     2401 aaggatccag agccatcgtg gcttttgaca atatctccat cagcctggac tgctacctca
     2461 ccattagcgg agaggacaag atcctgcaga atacagcacc caaatcaaga aacctgtttg
     2521 agagaaaccc aaacaaggag ctgaaacccg gggaaaattc accaagacag acccccatct
     2581 ttgaccctac agttcattgg ctgttcacca catgtggggc cagcgggccc catggcccca
     2641 cccaggcaca gtgcaacaac gcctaccaga actccaacct gagcgtggag gtggggagcg
     2701 agggccccct gaaaggcatc cagatctgga aggtgccagc caccgacacc tacagcatct
     2761 cgggctacgg agctgctggc gggaaaggcg ggaagaacac catgatgcgg tcccacggcg
     2821 tgtctgtgct gggcatcttc aacctggaga aggatgacat gctgtacatc ctggttgggc
     2881 agcagggaga ggacgcctgc cccagtacaa accagttaat ccagaaagtc tgcattggag
     2941 agaacaatgt gatagaagaa gaaatccgtg tgaacagaag cgtgcatgag tgggcaggag
     3001 gcggaggagg agggggtgga gccacctacg tatttaagat gaaggatgga gtgccggtgc
     3061 ccctgatcat tgcagccgga ggtggcggca gggcctacgg ggccaagaca gacacgttcc
     3121 acccagagag actggagaat aactcctcgg ttctagggct aaacggcaat tccggagccg
     3181 caggtggtgg aggtggctgg aatgataaca cttccttgct ctgggccgga aaatctttgc
     3241 aggagggtgc caccggagga cattcctgcc cccaggccat gaagaagtgg gggtgggaga
     3301 caagaggggg tttcggaggg ggtggagggg ggtgctcctc aggtggagga ggcggaggat
     3361 atataggcgg caatgcagcc tcaaacaatg accccgaaat ggatggggaa gatggggttt
     3421 ccttcatcag tccactgggc atcctgtaca ccccagcttt aaaagtgatg gaaggccacg
     3481 gggaagtgaa tattaagcat tatctaaact gcagtcactg tgaggtagac gaatgtcaca
     3541 tggaccctga aagccacaag gtcatctgct tctgtgacca cgggacggtg ctggctgagg
     3601 atggcgtctc ctgcattgtg tcacccaccc cggagccaca cctgccactc tcgctgatcc
     3661 tctctgtggt gacctctgcc ctcgtggccg ccctggtcct ggctttctcc ggcatcatga
     3721 ttgtgtaccg ccggaagcac caggagctgc aagccatgca gatggagctg cagagccctg
     3781 agtacaagct gagcaagctc cgcacctcga ccatcatgac cgactacaac cccaactact
     3841 gctttgctgg caagacctcc tccatcagtg acctgaagga ggtgccgcgg aaaaacatca
     3901 ccctcattcg gggtctgggc catggcgcct ttggggaggt gtatgaaggc caggtgtccg
     3961 gaatgcccaa cgacccaagc cccctgcaag tggctgtgaa gacgctgcct gaagtgtgct
     4021 ctgaacagga cgaactggat ttcctcatgg aagccctgat catcagcaaa ttcaaccacc
     4081 agaacattgt tcgctgcatt ggggtgagcc tgcaatccct gccccggttc atcctgctgg
     4141 agctcatggc ggggggagac ctcaagtcct tcctccgaga gacccgccct cgcccgagcc
     4201 agccctcctc cctggccatg ctggaccttc tgcacgtggc tcgggacatt gcctgtggct
     4261 gtcagtattt ggaggaaaac cacttcatcc accgagacat tgctgccaga aactgcctct
     4321 tgacctgtcc aggccctgga agagtggcca agattggaga cttcgggatg gcccgagaca
     4381 tctacagggc gagctactat agaaagggag gctgtgccat gctgccagtt aagtggatgc
     4441 ccccagaggc cttcatggaa ggaatattca cttctaaaac agacacatgg tcctttggag
     4501 tgctgctatg ggaaatcttt tctcttggat atatgccata ccccagcaaa agcaaccagg
     4561 aagttctgga gtttgtcacc agtggaggcc ggatggaccc acccaagaac tgccctgggc
     4621 ctgtataccg gataatgact cagtgctggc aacatcagcc tgaagacagg cccaactttg
     4681 ccatcatttt ggagaggatt gaatactgca cccaggaccc ggatgtaatc aacaccgctt
     4741 tgccgataga atatggtcca cttgtggaag aggaagagaa agtgcctgtg aggcccaagg
     4801 accctgaggg ggttcctcct ctcctggtct ctcaacaggc aaaacgggag gaggagcgca
     4861 gcccagctgc cccaccacct ctgcctacca cctcctctgg caaggctgca aagaaaccca
     4921 cagctgcaga ggtctctgtt cgagtcccta gagggccggc cgtggaaggg ggacacgtga
     4981 atatggcatt ctctcagtcc aaccctcctt cggagttgca caaggtccac ggatccagaa
     5041 acaagcccac cagcttgtgg aacccaacgt acggctcctg gtttacagag aaacccacca
     5101 aaaagaataa tcctatagca aagaaggagc cacacgacag gggtaacctg gggctggagg
     5161 gaagctgtac tgtcccacct aacgttgcaa ctgggagact tccgggggcc tcactgctcc
     5221 tagagccctc ttcgctgact gccaatatga aggaggtacc tctgttcagg ctacgtcact
     5281 tcccttgtgg gaatgtcaat tacggctacc agcaacaggg cttgccctta gaagccgcta
     5341 ctgcccctgg agctggtcat tacgaggata ccattctgaa aagcaagaat agcatgaacc
     5401 agcctgggcc cccaactttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
     5461 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     5521 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     5581 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgcctcctct
     5641 agaccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc gggtgacgca
     5701 caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg gttcgtaagc
     5761 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     5821 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt
     5881 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat
     5941 ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacattatga
     6001 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
     6061 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     6121 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     6181 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     6241 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     6301 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     6361 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     6421 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     6481 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     6541 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     6601 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     6661 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc
     6721 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg
     6781 gcaaataacc ctcgagccac ccatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     6841 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6901 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6961 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7021 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7081 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7141 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7201 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7261 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7321 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7381 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7441 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7501 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7561 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     7621 gcagcgagtc agtgagcgag gaagcggaag
//
