LOCUS       sequence-298118-        5475 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 5475)
  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 5475)
  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-AXL
FEATURES             Location/Qualifiers
     source          1..5475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3354..3453)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3470..3489)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3471..3489)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3494..3510)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3494..3510)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3941..4732
                     /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      4825..5413
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5314..5333
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcacc atggcgtggc ggtgccccag gatgggcagg gtcccgctgg cctggtgctt
      721 ggcgctgtgc ggctgggcgt gcatggcccc caggggcacg ccggctgaag aaagtccctt
      781 cgtgggcaac ccagggaata tcacaggtgc ccggggactc acgggcaccc ttcggtgtca
      841 gctccaggtt cagggagagc cccccgaggt acattggctt cgggatggac agatcctgga
      901 gctcgcggac agcacccaga cccaggtgcc cctgggtgag gatgaacagg atgactggat
      961 agtggtcagc cagctcagaa tcacctccct gcagctttcc gacacgggac agtaccagtg
     1021 tttggtgttt ctgggacatc agaccttcgt gtcccagcct ggctatgttg ggctggaggg
     1081 cttgccttac ttcctggagg agcccgaaga caggactgtg gccgccaaca cccccttcaa
     1141 cctgagctgc caagctcagg gacccccaga gcccgtggac ctactctggc tccaggatgc
     1201 tgtccccctg gccacggctc caggtcacgg cccccagcgc agcctgcatg ttccagggct
     1261 gaacaagaca tcctctttct cctgcgaagc ccataacgcc aagggggtca ccacatcccg
     1321 cacagccacc atcacagtgc tcccccagca gccccgtaac ctccacctgg tctcccgcca
     1381 acccacggag ctggaggtgg cttggactcc aggcctgagc ggcatctacc ccctgaccca
     1441 ctgcaccctg caggctgtgc tgtcagacga tgggatgggc atccaggcgg gagaaccaga
     1501 ccccccagag gagcccctca cctcgcaagc atccgtgccc ccccatcagc ttcggctagg
     1561 cagcctccat cctcacaccc cttatcacat ccgcgtggca tgcaccagca gccagggccc
     1621 ctcatcctgg acccactggc ttcctgtgga gacgccggag ggagtgcccc tgggcccccc
     1681 tgagaacatt agtgctacgc ggaatgggag ccaggccttc gtgcattggc aagagccccg
     1741 ggcgcccctg cagggtaccc tgttagggta ccggctggcg tatcaaggcc aggacacccc
     1801 agaggtgcta atggacatag ggctaaggca agaggtgacc ctggagctgc agggggacgg
     1861 gtctgtgtcc aatctgacag tgtgtgtggc agcctacact gctgctgggg atggaccctg
     1921 gagcctccca gtacccctgg aggcctggcg cccagggcaa gcacagccag tccaccagct
     1981 ggtgaaggaa ccttcaactc ctgccttctc gtggccctgg tggtatgtac tgctaggagc
     2041 agtcgtggcc gctgcctgtg tcctcatctt ggctctcttc cttgtccacc ggcgaaagaa
     2101 ggagacccgt tatggagaag tgtttgaacc aacagtggaa agaggtgaac tggtagtcag
     2161 gtaccgcgtg cgcaagtcct acagtcgtcg gaccactgaa gctaccttga acagcttggg
     2221 catcagtgaa gagctgaagg agaagctgcg ggatgtgatg gtggaccggc acaaggtggc
     2281 cctggggaag actctgggag agggagagtt tggagctgtg atggaaggcc agctcaacca
     2341 ggacgactcc atcctcaagg tggctgtgaa gacgatgaag attgccatct gcacgaggtc
     2401 agagctggag gatttcctga gtgaagcggt ctgcatgaag gaatttgacc atcccaacgt
     2461 catgaggctc atcggtgtct gtttccaggg ttctgaacga gagagcttcc cagcacctgt
     2521 ggtcatctta cctttcatga aacatggaga cctacacagc ttcctcctct attcccggct
     2581 cggggaccag ccagtgtacc tgcccactca gatgctagtg aagttcatgg cagacatcgc
     2641 cagtggcatg gagtatctga gtaccaagag attcatacac cgggacctgg cggccaggaa
     2701 ctgcatgctg aatgagaaca tgtccgtgtg tgtggcggac ttcgggctct ccaagaagat
     2761 ctacaatggg gactactacc gccagggacg tatcgccaag atgccagtca agtggattgc
     2821 cattgagagt ctagctgacc gtgtctacac cagcaagagc gatgtgtggt ccttcggggt
     2881 gacaatgtgg gagattgcca caagaggcca aaccccatat ccgggcgtgg agaacagcga
     2941 gatttatgac tatctgcgcc ggggaaatcg cctgaagcag cctgcggact gtctggatgg
     3001 actgtatgcc ttgatgtcgc ggtgctggga gctaaatccc caggaccggc caagttttac
     3061 agagctgcgg gaagatttgg agaacacact gaaggccttg cctcctgccc aggagcctga
     3121 cgaaatcctc tatgtcaaca tggatgaggg tggaggttat cctgaacccc ctggagctgc
     3181 aggaggagct gaccccccaa cccagccaga ccctaaggat tcctgtagct gcctcactgc
     3241 ggctgaggtc catcctgctg gacgctatgt cctctgccct tccacaaccc ctagccccgc
     3301 tcagcctgct gataggggct ccccagcagc cccagggcag gaggatggtg cctagccaac
     3361 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     3421 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     3481 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     3541 atgttacatt gcacaagata aaaatatatc atcatgcctc ctctagacca gccaggacag
     3601 aaatgcctcg acttcgctgc tacccaaggt tgccgggtga cgcacaccgt ggaaacggat
     3661 gaaggcacga acccagtgga cataagcctg ttcggttcgt aagctgtaat gcaagtagcg
     3721 tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt
     3781 ggcggttttc atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc
     3841 caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt
     3901 tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc
     3961 cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac
     4021 gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga
     4081 tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat
     4141 caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga
     4201 agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact
     4261 gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat
     4321 cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg
     4381 tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa
     4441 tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt
     4501 gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt
     4561 cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc
     4621 tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga
     4681 agaatttgtc cactacgtga aaggcgagat caccaaggta gtcggcaaat aaccctcgag
     4741 ccacccatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     4801 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4861 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4921 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4981 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5041 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5101 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5161 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5221 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5281 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5341 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5401 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5461 ttttgctcac atgtt
//
