LOCUS       sequence-232057-        5256 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 5256)
  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 5256)
  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-NTRK2
FEATURES             Location/Qualifiers
     source          1..5256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(154..181)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(273..359)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     408..430
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     422..439
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     423..439
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    455..554
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3021..3120)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3137..3156)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3138..3156)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3161..3177)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3161..3177)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3608..4399
                     /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      4492..5080
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4981..5000
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5234..5251
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
       61 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg
      121 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
      181 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
      241 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
      301 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
      361 tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg
      421 ttgtaaaacg acggccagtc ttaagctcgg gccccaaata atgattttat tttgactgat
      481 agtgacctgt tcgttgcaac aaattgatga gcaatgcttt tttataatgc caactttgta
      541 caaaaaagtt ggcaccatgt cgtcctggat aaggtggcat ggacccgcca tggcgcggct
      601 ctggggcttc tgctggctgg ttgtgggctt ctggagggcc gctttcgcct gtcccacgtc
      661 ctgcaaatgc agtgcctctc ggatctggtg cagcgaccct tctcctggca tcgtggcatt
      721 tccgagattg gagcctaaca gtgtagatcc tgagaacatc accgaaattt tcatcgcaaa
      781 ccagaaaagg ttagaaatca tcaacgaaga tgatgttgaa gcttatgtgg gactgagaaa
      841 tctgacaatt gtggattctg gattaaaatt tgtggctcat aaagcatttc tgaaaaacag
      901 caacctgcag cacatcaatt ttacccgaaa caaactgacg agtttgtcta ggaaacattt
      961 ccgtcacctt gacttgtctg aactgatcct ggtgggcaat ccatttacat gctcctgtga
     1021 cattatgtgg atcaagactc tccaagaggc taaatccagt ccagacactc aggatttgta
     1081 ctgcctgaat gaaagcagca agaatattcc cctggcaaac ctgcagatac ccaattgtgg
     1141 tttgccatct gcaaatctgg ccgcacctaa cctcactgtg gaggaaggaa agtctatcac
     1201 attatcctgt agtgtggcag gtgatccggt tcctaatatg tattgggatg ttggtaacct
     1261 ggtttccaaa catatgaatg aaacaagcca cacacagggc tccttaagga taactaacat
     1321 ttcatccgat gacagtggga agcagatctc ttgtgtggcg gaaaatcttg taggagaaga
     1381 tcaagattct gtcaacctca ctgtgcattt tgcaccaact atcacatttc tcgaatctcc
     1441 aacctcagac caccactggt gcattccatt cactgtgaaa ggcaacccca aaccagcgct
     1501 tcagtggttc tataacgggg caatattgaa tgagtccaaa tacatctgta ctaaaataca
     1561 tgttaccaat cacacggagt accacggctg cctccagctg gataatccca ctcacatgaa
     1621 caatggggac tacactctaa tagccaagaa tgagtatggg aaggatgaga aacagatttc
     1681 tgctcacttc atgggctggc ctggaattga cgatggtgca aacccaaatt atcctgatgt
     1741 aatttatgaa gattatggaa ctgcagcgaa tgacatcggg gacaccacga acagaagtaa
     1801 tgaaatccct tccacagacg tcactgataa aaccggtcgg gaacatctct cggtctatgc
     1861 tgtggtggtg attgcgtctg tggtgggatt ttgccttttg gtaatgctgt ttctgcttaa
     1921 gttggcaaga cactccaagt ttggcatgaa aggcccagcc tccgttatca gcaatgatga
     1981 tgactctgcc agcccactcc atcacatctc caatgggagt aacactccat cttcttcgga
     2041 aggtggccca gatgctgtca ttattggaat gaccaagatc cctgtcattg aaaatcccca
     2101 gtactttggc atcaccaaca gtcagctcaa gccagacaca tttgttcagc acatcaagcg
     2161 acataacatt gttctgaaaa gggagctagg cgaaggagcc tttggaaaag tgttcctagc
     2221 tgaatgctat aacctctgtc ctgagcagga caagatcttg gtggcagtga agaccctgaa
     2281 ggatgccagt gacaatgcac gcaaggactt ccaccgtgag gccgagctcc tgaccaacct
     2341 ccagcatgag cacatcgtca agttctatgg cgtctgcgtg gagggcgacc ccctcatcat
     2401 ggtctttgag tacatgaagc atggggacct caacaagttc ctcagggcac acggccctga
     2461 tgccgtgctg atggctgagg gcaacccgcc cacggaactg acgcagtcgc agatgctgca
     2521 tatagcccag cagatcgccg cgggcatggt ctacctggcg tcccagcact tcgtgcaccg
     2581 cgatttggcc accaggaact gcctggtcgg ggagaacttg ctggtgaaaa tcggggactt
     2641 tgggatgtcc cgggacgtgt acagcactga ctactacagg gtcggtggcc acacaatgct
     2701 gcccattcgc tggatgcctc cagagagcat catgtacagg aaattcacga cggaaagcga
     2761 cgtctggagc ctgggggtcg tgttgtggga gattttcacc tatggcaaac agccctggta
     2821 ccagctgtca aacaatgagg tgatagagtg tatcactcag ggccgagtcc tgcagcgacc
     2881 ccgcacgtgc ccccaggagg tgtatgagct gatgctgggg tgctggcagc gagagcccca
     2941 catgaggaag aacatcaagg gcatccatac cctccttcag aacttggcca aggcatctcc
     3001 ggtctacctg gacattctag gcccaacttt cttgtacaaa gttggcatta taagaaagca
     3061 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     3121 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     3181 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     3241 atgcctcctc tagaccagcc aggacagaaa tgcctcgact tcgctgctac ccaaggttgc
     3301 cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat aagcctgttc
     3361 ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac
     3421 cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt
     3481 tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg
     3541 tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt
     3601 aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg
     3661 cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt
     3721 ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct
     3781 tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg
     3841 agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc
     3901 gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct
     3961 tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc
     4021 aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc
     4081 tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga
     4141 ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt
     4201 aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc
     4261 ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg
     4321 cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac
     4381 caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta acgtgagtta
     4441 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4501 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4561 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4621 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4681 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4741 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4801 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4861 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4921 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4981 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5041 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5101 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5161 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     5221 aacgaccgag cgcagcgagt cagtgagcga ggaagc
//
