LOCUS       sequence-424758-        7152 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 7152)
  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 7152)
  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-LMTK2
FEATURES             Location/Qualifiers
     source          1..7152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(70..87)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(70..86)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(79..101)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      150..236
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      328..355
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(514..531)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(685..1273)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(765..784)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1366..2157)
                     /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     2588..2604
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2588..2604
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2609..2628
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2609..2627
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2645..2744
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(join(7107..7152,1..54))
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttggggccc
       61 gagcttaaga ctggccgtcg ttttacaacg tcgtgactgg gaaaacatcc atgctagcgt
      121 taacgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcggaagact
      181 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
      241 cgggagcgga tttgaacgtt gtgaagcaac ggcccggagg gtggcgggca ggacgcccgc
      301 cataaactgc caggcatcaa actaagcaga aggccatcct gacggatggc ctttttgcgt
      361 ttctacaaac tcttcctggc tagcggtacg cgtattaatt gcgttgcgct cactgcccgc
      421 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
      481 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
      541 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
      601 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
      661 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
      721 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      781 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
      841 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
      901 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
      961 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1021 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1081 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     1141 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1201 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1261 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1321 cgcgtaactc acgttaaggg attttggtca tgggtggctc gagggttatt tgccgactac
     1381 cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc
     1441 caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata
     1501 ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc
     1561 ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc
     1621 ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg
     1681 ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc
     1741 tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc
     1801 tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg
     1861 ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct
     1921 ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc
     1981 atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc
     2041 atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat
     2101 gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcataat
     2161 gtttaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tccataacat
     2221 caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac
     2281 cccaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg
     2341 ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagcttacg
     2401 aaccgaacag gcttatgtcc actgggttcg tgccttcatc cgtttccacg gtgtgcgtca
     2461 cccggcaacc ttgggtagca gcgaagtcga ggcatttctg tcctggctgg tctagaggag
     2521 gcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacgggcca
     2581 gagctgccag gaaacagcta tgaccatgta atacgactca ctatagggga tatcagctgg
     2641 atggcaaata atgattttat tttgactgat agtgacctgt tcgttgcaac aaattgataa
     2701 gcaatgcttt cttataatgc caactttgta caagaaagtt ggggtgtgtg agggaaaagc
     2761 tggcaatgtt gacgggagag aaggcttgga gctgagcagg ttacttgttg tctttgacat
     2821 aaaaggatct ggggagaagt catcatccca ctcaaagcca ccaccttctt cgtcggtgga
     2881 gctttcggag ttgatgcacc tgctcaggta gggagagcct gaggctgggg ctgggccgct
     2941 caggtccggg ccgcacgcct ctcctccaca gggccccagc tctttggttg gggtctcctg
     3001 gtcaaacagg tagactgtga catcatcgaa aaacgtgact gccttctttt ccttcttcca
     3061 gtcctcgggc agtgggtcgg gggcattggc cgctgtgggc ttcaacagac tccgcaggtg
     3121 ccttccgtcc tcgttgctga ggatgatggg cacggggtgc tcggtctcgt cctccgactc
     3181 ggagctgagg ctatgcaggt tgaaggcccg caggtcctcg tccgagtcgt cgctgttttc
     3241 gtcctcctcg tctgcatcca tcccgtcctc tgagaggtcg agcatccctg acaccccgag
     3301 tttccccagg tacttccctt cgatgtccgg ctccttcaac ttcgggccct cggagtggct
     3361 ggacagggac ttgtccagcg cagaattggt gtaggcaagg agttcgttcg tgttggtccc
     3421 cgtggaagcg agggtgcagg ggcgatcgtc ctgtgtctcg aagtcatcgc cgctgctaag
     3481 ttctgcattc agcacactcc agggactgct ggcctcgtct tccgtgggat gcacctgctg
     3541 gggaacacca gtctgtgctg gctccggcgt ggctcttaat tccaggtcac tggagttgtg
     3601 caaagcagac agacaacttt catctggctg gctctttctg gcttccaggc agctatcctg
     3661 ggcagcagga ctttgctcgg ggaggactgg ctcgggtagg ggctgctcct gtaccaacac
     3721 caaggcggat ggggaggttc ccgggacctc ctcagacctt ttctcgggct cagagtcatt
     3781 gtctgagaag tacgcagagt ctcggtatga accctgggag ccagctgtga acgtctcagg
     3841 ggtcacttct gtgcctctgt gaccatcgcc ggcatctgag atgacaatga ccgggttggg
     3901 aggcagaccg ctgtggccgc catcgcccgt ggtggctggt tctgagtctg cggtgccctc
     3961 gggagcggga tgcaaggtcc actcgggaga ctccaagttc tctgtttcgt agccactgtc
     4021 tgccggctta tcggggggca ctagtttctg gggagtgtga gatcctaaag agtccagtag
     4081 cgcttcgtgg acatccactg actccagaga gtccggggtt tccagggacg gctcagaggc
     4141 tgggaagggt gctgacaagc tgtcctccag gaggctgtcc tgactggttg agtccgagga
     4201 gagggcagac accaagcctg cttcttttgc tgcgtcttta gaattgagct gatttaaagt
     4261 ctccacagcc tctaagtttt tctctagccg gcgatgactg tgatggtctt ccgaaaatgg
     4321 tttattgtgc aattcctgtc ccagttccgg gagactactc cctacactca ccctgctggc
     4381 aaggatgtca tcagcaagaa cagagtcact ttcggtgagt cgcagggtct cctcactctc
     4441 gccaggagag tcctgggacc tgttatccag gctgtgggtt ctggcatcag ttgagagaat
     4501 ttcaaccggg acagtcacag cgtctggact gatgtcctgg tggagacagt cctccgggac
     4561 aataacatct aaacacgtgg gctgggtttc tccctgtgtt gggagtgagt ctgggggtac
     4621 ccgacggggc gtttcttctg tttcgaagga ggtaggaggt acctgcactt ctggggaaga
     4681 ctgcaatgag gtgctcaaag tgtcacctgt gagcatgaca tctgtatcgt caccctttgt
     4741 gcttacgttt tcctgcaaca aagacatgcc tggcttattt tcagcaaact gaagctcagt
     4801 atctaaagag tttctacatg acgtttctaa catagcttca gtaaaaccag cattcttaag
     4861 ttctgtctga agatcattta tgtgttcttt gctggacaat gagcctttta gtaagttttt
     4921 ctcttgaaga aataaaaagt tttctgacaa ttcttgaaca ttcaatggat caaaattatc
     4981 ttggtgcata agattatctg ataggcagag aggctcactg tcaaaaatct tacggggctt
     5041 ttctttctca aagtggcctg ttatgactga ctctttgggg ttatccaaac tggaacttaa
     5101 agtggcaggt ttaaagtcgg cttgaactcc gtttaattcc attaagtcga atattttttg
     5161 gtgactgggc aaatcttccg atttgtccac atcattaaat atattgttga aagggctctc
     5221 ggggccactg gtcacgtgta agtcccctgg taagctagag tctttggggt ctgtactgct
     5281 ttggaagaag tcctcatctg tactggactc ctcaagttca acgctcctga gcgccgtgag
     5341 ctgggacagt tcagggccag tcctttctgg attatccatg tcggttgtga gcagcgctgg
     5401 tgggtaatca agctccaagt tactaccact tttttcttct aactggatat aatagtcgct
     5461 tccaacagaa aggttgtggg catcaaaaac aggaaccact ccagggaccc tcagggggac
     5521 atcctggccg ctgtcatctt gctttccggg cccaggatct gaaagcgaac tctcaaaaac
     5581 ttcaaccgga tagaagatgc tcgtgtagga caagccttcg tccaggtggc cccggctgcg
     5641 ctcgtcaaag tggtcgtgct tagcggcctc ccagacatac tcgaagctca ggccctggct
     5701 ggtttcggtc acggtgagga cttcctccat ttcacgaccc agccggtccc tggcaaagtg
     5761 gtcgagaatt gggaatgcag cattgttgga ggagtctctg ctgtttgtgt tcggcttcag
     5821 agcgttccac tgctgttcaa agtcgacctc tgagtcccgc tggctctgca gccgcaggta
     5881 agtcagcagc ctgtgcacat cttcagccgc gggtctcttt tctggtgaca gccaacagaa
     5941 ctgtaagact tcataccatc tatcagagta gggctgctcc agctggggct tcgggagttt
     6001 tgtgtctctc tctctaatga cttggttgag gacatctaag ttggaaaggt ttgaatacgg
     6061 ctgtgcggca ttgtcaaaaa gctcccaaag tgtcacaccc agagaccaga tattactata
     6121 cttagtctga tctgcagtta gcagtctgtc ttgaaagctg gttactaatt ctggagcagt
     6181 ccatcgcaga gggaaaactt ttttatcatc tgtttcaata taatcctcct tgtacctgct
     6241 gaatcctatt ccgtaatctc ccactttcac atttaagtcg gaggtgagaa aacaattccg
     6301 cagggctaaa tcactgtgca ggaagtgcag cttgtgcatg gcggccagcc ccgcggcgac
     6361 ctcgcacgcc atcctctgca gcagcatggt ctgtgagtcc ccccgcatgt gctcctgctc
     6421 gctgcgcaga tacgccttca ggtcacccaa gtcacagaac tcaaacacca ggaggtaggg
     6481 aatcgcttct acgcactgtc caacacactg aagaatattt ggatgctgaa gaatgtagta
     6541 aggttctcca tttttcaaaa aagtatcttg ttcctttggg ttggcacttg cttttaactc
     6601 cttcacgatg actcttgcta cgctagtgcc cgtgtaaatc tctcccaaga gaacctttcc
     6661 aaaccagcca tttccaattt cctgtatgta gtttagactg tggcgggcaa cttgagactt
     6721 caatccctct acagaaggct ggaattgcga gggagctggg agtgaaatat ttggtactga
     6781 aagtgtgaag acctctgctg gggactgaac agagggagtg tcttctgctg gtggtgtgaa
     6841 atctatctca tcatcaaaat tatcttcaaa ttccttaaag tctatttctg ggtccttaca
     6901 gcaggataca cagtttgcaa ttaacactat taatattatt aaactgcaca cacacaggat
     6961 cacaaaagat gaggaaactt ctgcagcagg tggcgcctcc cctgcacctg tttgcggaag
     7021 tggcgcggcc ccagcactgc cggcgatcag gaggaccagc agcagcagca gcagcctccg
     7081 ccgcaacgcc ggcggccccg gcatggtgcc aacttttttg tacaaagttg gcattataaa
     7141 aaagcattgc tc
//
