LOCUS       sequence-159438-        5742 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 5742)
  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 5742)
  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-EPHB1
FEATURES             Location/Qualifiers
     source          1..5742
                     /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     417..434
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     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(3502..3601)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3618..3637)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3619..3637)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3642..3658)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3642..3658)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4089..4880
                     /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      4973..5561
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5462..5481
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5715..5732
                     /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 aagttggcat ggccctggat tatctactac tgctcctcct ggcatccgca gtggctgcga
      601 tggaagaaac gttaatggac accagaacgg ctactgcaga gctgggctgg acggccaatc
      661 ctgcgtccgg gtgggaagaa gtcagtggct acgatgaaaa cctgaacacc atccgcacct
      721 accaggtgtg caatgtcttc gagcccaacc agaacaattg gctgctcacc accttcatca
      781 accggcgggg ggcccatcgc atctacacag agatgcgctt cactgtgaga gactgcagca
      841 gcctccctaa tgtcccagga tcctgcaagg agaccttcaa cttgtattac tatgagactg
      901 actctgtcat tgccaccaag aagtcagcct tctggtctga ggccccctac ctcaaagtag
      961 acaccattgc tgcagatgag agcttctccc aggtggactt tgggggaagg ctgatgaagg
     1021 taaacacaga agtcaggagc tttgggcctc ttactcggaa tggtttttac ctcgcttttc
     1081 aggattatgg agcctgtatg tctcttcttt ctgtccgtgt cttcttcaaa aagtgtccca
     1141 gcattgtgca aaattttgca gtgtttccag agactatgac aggggcagag agcacatctc
     1201 tggtgattgc tcggggcaca tgcatcccca acgcagagga agtggacgtg cccatcaaac
     1261 tctactgcaa cggggatggg gaatggatgg tgcctattgg gcgatgcacc tgcaagcctg
     1321 gctatgagcc tgagaacagc gtggcatgca aggcttgccc tgcagggaca ttcaaggcca
     1381 gccaggaagc tgaaggctgc tcccactgcc cctccaacag ccgctcccct gcagaggcgt
     1441 ctcccatctg cacctgtcgg accggttatt accgagcgga ctttgaccct ccagaagtgg
     1501 catgcactag cgtcccatca ggtccccgca atgttatctc catcgtcaat gagacgtcca
     1561 tcattctgga gtggcaccct ccaagggaga caggtgggcg ggatgatgtg acctacaaca
     1621 tcatctgcaa aaagtgccgg gcagaccgcc ggagctgctc ccgctgtgac gacaatgtgg
     1681 agtttgtgcc caggcagctg ggcctgacgg agtgccgcgt ctccatcagc agcctgtggg
     1741 cccacacccc ctacaccttt gacatccagg ccatcaatgg agtctccagc aagagtccct
     1801 tccccccaca gcacgtctct gtcaacatca ccacaaacca agccgccccc tccaccgttc
     1861 ccatcatgca ccaagtcagt gccactatga ggagcatcac cttgtcatgg ccacagccgg
     1921 agcagcccaa tggcatcatc ctggactatg agatccggta ctatgagaag gaacacaatg
     1981 agttcaactc ctccatggcc aggagtcaga ccaacacagc aaggattgat gggctgcggc
     2041 ctggcatggt atatgtggta caggtgcgtg cccgcactgt tgctggctac ggcaagttca
     2101 gtggcaagat gtgcttccag actctgactg acgatgatta caagtcagag ctgagggagc
     2161 agctgcccct gattgctggc tcggcagcgg ccggggtcgt gttcgttgtg tccttggtgg
     2221 ccatctctat cgtctgtagc aggaaacggg cttatagcaa agaggctgtg tacagcgata
     2281 agctccagca ttacagcaca ggccgaggct ccccagggat gaagatctac attgacccct
     2341 tcacttatga ggatcccaac gaagctgtcc gggagtttgc caaggagatt gatgtatctt
     2401 ttgtgaaaat tgaagaggtc atcggagcag gggagtttgg agaagtgtac aaggggcgtt
     2461 tgaaactgcc aggcaagagg gaaatctacg tggccatcaa gaccctgaag gcagggtact
     2521 cggagaagca gcgtcgggac tttctgagtg aggcgagcat catgggccag ttcgaccatc
     2581 ctaacatcat tcgcctggag ggtgtggtca ccaagagtcg gcctgtcatg atcatcacag
     2641 agttcatgga gaatggtgca ttggattctt tcctcaggca aaatgacggg cagttcaccg
     2701 tgatccagct tgtgggtatg ctcaggggca tcgctgctgg catgaagtac ctggctgaga
     2761 tgaattatgt gcatcgggac ctggctgcta ggaacattct ggtcaacagt aacctggtgt
     2821 gcaaggtgtc cgactttggc ctctcccgct acctccagga tgacacctca gatcccacct
     2881 acaccagctc cttgggaggg aagatccctg tgagatggac agctccagag gccatcgcct
     2941 accgcaagtt cacttcagcc agcgacgttt ggagctatgg gatcgtcatg tgggaagtca
     3001 tgtcatttgg agagagaccc tattgggata tgtccaacca agatgtcatc aatgccatcg
     3061 agcaggacta ccggctgccc ccacccatgg actgtccagc tgctctacac cagctcatgc
     3121 tggactgttg gcagaaggac cggaacagcc ggccccggtt tgcggagatt gtcaacaccc
     3181 tagataagat gatccggaac ccggcaagtc tcaagactgt ggcaaccatc accgccgtgc
     3241 cttcccagcc cctgctcgac cgctccatcc cagacttcac ggcctttacc accgtggatg
     3301 actggctcag cgccatcaaa atggtccagt acagggacag cttcctcact gctggcttca
     3361 cctccctcca gctggtcacc cagatgacat cagaagacct cctgagaata ggcatcacct
     3421 tggcaggcca tcagaagaag atcctgaaca gcattcattc tatgagggtc cagataagtc
     3481 agtcaccaac ggcaatggca ttgccaactt tcttgtacaa agttggcatt ataagaaagc
     3541 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
     3601 gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc
     3661 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     3721 catgcctcct ctagaccagc caggacagaa atgcctcgac ttcgctgcta cccaaggttg
     3781 ccgggtgacg cacaccgtgg aaacggatga aggcacgaac ccagtggaca taagcctgtt
     3841 cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga
     3901 ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt
     3961 ttggggtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat
     4021 gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt
     4081 taaacattat gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg
     4141 gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag
     4201 tggatggcgg cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc
     4261 ttgatgaaac aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg
     4321 gagagagcga gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc
     4381 cgtggcgtta tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc
     4441 ttgcaggtat cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag
     4501 caagagaaca tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc
     4561 ctgaacagga tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg
     4621 actgggctgg cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag
     4681 taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg
     4741 cccagtatca gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc
     4801 gcttggcctc gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca
     4861 ccaaggtagt cggcaaataa ccctcgagcc acccatgacc aaaatccctt aacgtgagtt
     4921 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4981 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5041 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5101 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5161 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5221 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5281 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5341 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5401 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5461 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5521 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5581 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5641 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     5701 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga ag
//
