LOCUS       sequence-181500-        4494 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 4494)
  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 4494)
  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-DYRK3
FEATURES             Location/Qualifiers
     source          1..4494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     141..158
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(317..344)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(436..522)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     571..593
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     585..602
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     586..602
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    618..717
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2422..2521)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2538..2557)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2539..2557)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2562..2578)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2562..2578)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3009..3800
                     /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      3893..4481
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4382..4401
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
       61 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      121 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
      181 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
      241 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag
      301 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct
      361 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa
      421 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
      481 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
      541 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca
      601 gtcttaagct cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc
      661 aacaaattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gttggcatga
      721 agtggaaaga gaagttgggg gatggtgtct atgacacctt catgatgata gatgaaacca
      781 aatgtccccc ctgttcaaat gtactctgca atccttctga accacctcca cccagaagac
      841 taaatatgac cactgagcag tttacaggag atcatactca gcactttttg gatggaggtg
      901 agatgaaggt agaacagctg tttcaagaat ttggcaacag aaaatccaat actattcagt
      961 cagatggcat cagtgactct gaaaaatgct ctcctactgt ttctcagggt aaaagttcag
     1021 attgcttgaa tacagtaaaa tccaacagtt catccaaggc acccaaagtg gtgcctctga
     1081 ctccagaaca agccctgaag caatataaac accacctcac tgcctatgag aaactggaaa
     1141 taattaatta tccagaaatt tactttgtag gtccaaatgc caagaaaaga catggagtta
     1201 ttggtggtcc caataatgga gggtatgatg atgcagatgg ggcctatatt catgtacctc
     1261 gagaccatct agcttatcga tatgaggtgc tgaaaattat tggcaagggg agttttgggc
     1321 aggtggccag ggtctatgat cacaaacttc gacagtacgt ggccctaaaa atggtgcgca
     1381 atgagaagcg ctttcatcgt caagcagctg aggagatccg gattttggag catcttaaga
     1441 aacaggataa aactggtagt atgaacgtta tccacatgct ggaaagtttc acattccgga
     1501 accatgtttg catggccttt gaattgctga gcatagacct ttatgagctg attaaaaaaa
     1561 ataagtttca gggttttagc gtccagttgg tacacaagtt tgcccagtcc atcttgcaat
     1621 ctttggatgc cctccacaaa aataagatta ttcactgcga tctgaagcca gaaaacattc
     1681 tcctgaaaca ccacgggcgc agttcaacca aggtcattga ctttgggtcc agctgtttcg
     1741 agtaccagaa gctctacaca tatatccagt ctcggttcta cagagctcca gaaatcatct
     1801 taggaagccg ctacagcaca ccaattgaca tatggagttt tggctgcatc cttgcagaac
     1861 ttttaacagg acagcctctc ttccctggag aggatgaagg agaccagttg gcctgcatga
     1921 tggagcttct agggatgcca ccaccaaaac ttctggagca atccaaacgt gccaagtact
     1981 ttattaattc caagggcata ccccgctact gctctgtgac tacccaggca gatgggaggg
     2041 ttgtgcttgt ggggggtcgc tcacgtaggg gtaaaaagcg gggtccccca ggcagcaaag
     2101 actgggggac agcactgaaa gggtgtgatg actacttgtt tatagagttc ttgaaaaggt
     2161 gtcttcactg ggacccctct gcccgcttga ccccagctca agcattaaga cacccttgga
     2221 ttagcaagtc tgtccccaga cctctcacca ccatagacaa ggtgtcaggg aaacgggtag
     2281 ttaatcctgc aagtgctttc cagggattgg gttctaagct gcctccagtt gttggaatag
     2341 ccaataagct taaagctaac ttaatgtcag aaaccaatgg tagtataccc ctatgcagtg
     2401 tattgccaaa actgattagc tacccaactt tcttgtacaa agttggcatt ataagaaagc
     2461 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
     2521 gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc
     2581 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     2641 catgcctcct ctagaccagc caggacagaa atgcctcgac ttcgctgcta cccaaggttg
     2701 ccgggtgacg cacaccgtgg aaacggatga aggcacgaac ccagtggaca taagcctgtt
     2761 cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga
     2821 ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt
     2881 ttggggtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat
     2941 gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt
     3001 taaacattat gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg
     3061 gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag
     3121 tggatggcgg cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc
     3181 ttgatgaaac aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg
     3241 gagagagcga gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc
     3301 cgtggcgtta tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc
     3361 ttgcaggtat cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag
     3421 caagagaaca tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc
     3481 ctgaacagga tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg
     3541 actgggctgg cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag
     3601 taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg
     3661 cccagtatca gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc
     3721 gcttggcctc gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca
     3781 ccaaggtagt cggcaaataa ccctcgagcc acccatgacc aaaatccctt aacgtgagtt
     3841 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3901 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3961 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4021 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4081 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4141 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4201 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4261 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4321 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4381 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4441 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacg
//
