LOCUS       sequence-405246-        5307 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 5307)
  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 5307)
  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-NTRK3
FEATURES             Location/Qualifiers
     source          1..5307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(59..145)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     194..216
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     208..225
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     209..225
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    241..340
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2858..2957)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2974..2993)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2975..2993)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2998..3014)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2998..3014)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3445..4236
                     /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      4329..4917
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4818..4837
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5071..5088
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5247..5274)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
       61 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
      121 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
      181 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
      241 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
      301 tgctttttta taatgccaac tttgtacaaa aaagttggca ccatggatgt ctctctttgc
      361 ccagccaagt gtagtttctg gcggattttc ttgctgggaa gcgtctggct ggactatgtg
      421 ggctccgtgc tggcttgccc tgcaaattgt gtctgcagca agactgagat caattgccgg
      481 cggccggacg atgggaacct cttccccctc ctggaagggc aggattcagg gaacagcaat
      541 gggaacgcca gtatcaacat cacggacatc tcaaggaata tcacttccat acacatagag
      601 aactggcgca gtcttcacac gctcaacgcc gtggacatgg agctctacac cggacttcaa
      661 aagctgacca tcaagaactc aggacttcgg agcattcagc ccagagcctt tgccaagaac
      721 ccccatttgc gttatataaa cctgtcaagt aaccggctca ccacactctc gtggcagctc
      781 ttccagacgc tgagtcttcg ggaattgcag ttggagcaga actttttcaa ctgcagctgt
      841 gacatccgct ggatgcagct ctggcaggag cagggggagg ccaagctcaa cagccagaac
      901 ctctactgca tcaacgctga tggctcccag cttcctctct tccgcatgaa catcagtcag
      961 tgtgaccttc ctgagatcag cgtgagccac gtcaacctga ccgtacgaga gggtgacaat
     1021 gctgttatca cttgcaatgg ctctggatca ccccttcctg atgtggactg gatagtcact
     1081 gggctgcagt ccatcaacac tcaccagacc aatctgaact ggaccaatgt tcatgccatc
     1141 aacttgacgc tggtgaatgt gacgagtgag gacaatggct tcaccctgac gtgcattgca
     1201 gagaacgtgg tgggcatgag caatgccagt gttgccctca ctgtctacta tcccccacgt
     1261 gtggtgagcc tggaggagcc tgagctgcgc ctggagcact gcatcgagtt tgtggtgcgt
     1321 ggcaaccccc caccaacgct gcactggctg cacaatgggc agcctctgcg ggagtccaag
     1381 atcatccatg tggaatacta ccaagaggga gagatttccg agggctgcct gctcttcaac
     1441 aagcccaccc actacaacaa tggcaactat accctcattg ccaaaaaccc actgggcaca
     1501 gccaaccaga ccatcaatgg ccacttcctc aaggagccct ttccagagag cacggataac
     1561 tttatcttgt ttgacgaagt gagtcccaca cctcctatca ctgtgaccca caaaccagaa
     1621 gaagacactt ttggggtatc catagcagtt ggacttgctg ctcttgcctg tgtcctgttg
     1681 gtggttctct tcgtcatgat caacaaatat ggtcgacggt ccaaatttgg aatgaagggt
     1741 cccgtggctg tcatcagtgg tgaggaggac tcagccagcc cactgcacca catcaaccac
     1801 ggcatcacca cgccctcgtc actggatgcc gggcccgaca ctgtggtcat tggcatgact
     1861 cgcatccctg tcattgagaa cccccagtac ttccgtcagg gacacaactg ccacaagccg
     1921 gacacgtatg tgcagcacat taagaggaga gacatcgtgc tgaagcgaga actgggtgag
     1981 ggagcctttg gaaaggtctt cctggccgag tgctacaacc tcagcccgac caaggacaag
     2041 atgcttgtgg ctgtgaaggc cctgaaggat cccaccctgg ctgcccggaa ggatttccag
     2101 agggaggccg agctgctcac caacctgcag catgagcaca ttgtcaagtt ctatggagtg
     2161 tgcggcgatg gggaccccct catcatggtc tttgaataca tgaagcatgg agacctgaat
     2221 aagttcctca gggcccatgg gccagatgca atgatccttg tggatggaca gccacgccag
     2281 gccaagggtg agctggggcc ctcccaaatg ctccacattg ccagtcagat cgcctcgggt
     2341 atggtgtacc tggcctccca gcactttgtg caccgagacc tggccaccag gaactgcctg
     2401 gttggagcga atctgctagt gaagattggg gacttcggca tgtccagaga tgtctacagc
     2461 acggattatt acaggctctt taatccatct ggaaatgatt tttgtatatg gtgtgaggtg
     2521 ggaggacaca ccatgctccc cattcgctgg atgcctcctg aaagcatcat gtaccggaag
     2581 ttcactacag agagtgatgt atggagcttc ggggtgatcc tctgggagat cttcacctat
     2641 ggaaagcagc catggttcca actctcaaac acggaggtca ttgagtgcat tacccaaggt
     2701 cgtgttttgg agcggccccg agtctgcccc aaagaggtgt acgatgtcat gctggggtgc
     2761 tggcagaggg aaccacagca gcggttgaac atcaaggaga tctacaaaat cctccatgct
     2821 ttggggaagg ccaccccaat ctacctggac attcttggcc caactttctt gtacaaagtt
     2881 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
     2941 aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc
     3001 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     3061 gataaaaata tatcatcatg cctcctctag accagccagg acagaaatgc ctcgacttcg
     3121 ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag
     3181 tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac
     3241 tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct
     3301 tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt
     3361 tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt
     3421 cgccctaaaa caaagttaaa cattatgagg gaagcggtga tcgccgaagt atcgactcaa
     3481 ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat
     3541 ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt
     3601 acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa
     3661 acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg
     3721 cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg
     3781 cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct
     3841 atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa
     3901 ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta
     3961 tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc
     4021 atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca
     4081 atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt
     4141 ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac
     4201 gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct cgagccaccc atgaccaaaa
     4261 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4321 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4381 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4441 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4501 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4561 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4621 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4681 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4741 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4801 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4861 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4921 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4981 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     5041 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     5101 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     5161 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
     5221 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
     5281 tttgatgcct ggcagtttat ggcgggc
//
