LOCUS       sequence-357423-        5298 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 5298)
  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 5298)
  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-TNNI3K
FEATURES             Location/Qualifiers
     source          1..5298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2609..2708)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2725..2744)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2726..2744)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2749..2765)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2749..2765)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3196..3987
                     /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      4080..4668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4569..4588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4822..4839
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4998..5025)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5117..5203)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5252..5274
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5266..5283
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5267..5283
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagttggca ccatgggaaa ttataaatct
      121 agaccaaccc aaacttgtac tgatgaatgg aagaaaaaag tcagtgaatc atatgttatc
      181 acaatagaaa gattagaaga tgacctgcag atcaaggaaa aagaactgac agaactaagg
      241 aatatatttg gctctgatga agccttcagt aaagtcaatt taaattaccg cactgaaaat
      301 gggctgtctc tacttcattt atgttgcatt tgtggaggca agaaatcaca tattcgaact
      361 cttatgttga aagggctccg cccatctcga ctgacaagaa atggatttac agccttgcat
      421 ttagcagttt acaaggataa tgcagaattg atcacttctc tgcttcacag tggagctgat
      481 atacagcagg ttggatacgg tggcctcact gccctccata ttgctacaat agctggccac
      541 ctagaggctg ctgatgtgct gttgcaacat ggagctaatg tcaatattca agatgcagtt
      601 tttttcactc cattgcatat tgcagcgtac tatggacatg aacaggtaac tcgccttctt
      661 ttgaaatttg gtgctgatgt aaatgtaagt ggtgaagttg gagatagacc cctccaccta
      721 gcatctgcaa aaggattctt gaatattgca aaactcttga tggaagaagg cagcaaagca
      781 gatgtgaatg ctcaagataa tgaagaccat gtcccactcc atttctgttc tcgatttgga
      841 caccatgata tagttaagta tctgctgcaa agtgatttgg aagttcaacc tcatgttgtt
      901 aatatctatg gagatacccc cttacacctg gcatgctaca atggcaaatt tgaagttgcc
      961 aaggaaatca tccaaatatc aggaacagaa agtctgacta aggaaaacat cttcagtgaa
     1021 acagcttttc atagtgcttg tacctatggc aagagcattg acctagtcaa atttcttctt
     1081 gatcagaatg tcataaacat caaccaccaa ggaagggatg ggcacactgg attacactct
     1141 gcttgctacc acggtcacat tcgcctggtt cagttcttac tggataatgg agctgatatg
     1201 aatctagtgg cttgtgatcc cagcaggtct agtggtgaaa aagatgagca gacatgtttg
     1261 atgtgggctt atgaaaaagg gcatgatgcc attgtcacac tcctgaagca ttataagaga
     1321 ccacaagatg aattgccctg taatgaatat tctcagcctg gaggagatgg ctcctatgtg
     1381 tctgttccat cacccttggg gaagattaaa agcatgacaa aagagaaggc agatattctc
     1441 ctcctaagag ctggattgcc ttcacatttc catcttcagc tctcagaaat tgagttccat
     1501 gagattattg gctcaggttc ttttgggaaa gtatataaag gacgatgcag aaataaaata
     1561 gtggctataa aacgttatcg agccaatacc tactgctcca agtcagatgt ggatatgttt
     1621 tgccgagagg tgtccattct ctgccagctc aatcatccct gcgtaattca gtttgtgggt
     1681 gcttgcttga atgatcccag ccagtttgcc attgtcactc aatacatatc agggggttct
     1741 ctgttctccc tccttcatga gcagaagagg attcttgatt tgcagtctaa attaattatt
     1801 gcagtagatg ttgccaaagg catggagtac cttcacaacc tgacacagcc aattatacat
     1861 cgtgacttga acagtcacaa tattcttctc tatgaggatg ggcatgctgt ggtggcagat
     1921 tttggagaat caagatttct acagtctctg gatgaagaca acatgacaaa acaacctggg
     1981 aacctccgtt ggatggctcc tgaggtgttc acgcagtgca ctcggtacac catcaaagca
     2041 gatgtcttca gctatgctct gtgtctgtgg gaaattctca ctggcgaaat tccattcgct
     2101 catctcaagc cagcggctgc ggcagcagac atggcttacc accacatcag acctcccatt
     2161 ggctattcca ttcccaagcc catatcatct ctgctgatac gagggtggaa cgcatgtcct
     2221 gaaggaagac ccgaattttc tgaagttgtc atgaagttag aagagtgtct ctgcaacatt
     2281 gagctgatgt ctcctgcatc aagtaacagc agtgggtctc tctcaccttc ttcttcttct
     2341 gattgcctgg tgaaccgggg aggacctggc cggagtcatg tggcagcatt aagaagtcgt
     2401 ttcgaattgg aatatgctct aaatgcaagg tcctatgctg ctttgtccca aagtgctgga
     2461 caatattcct ctcaaggtct gtctttggag gagatgaaaa gaagtcttca atacacaccc
     2521 attgacaaat atggctatgt atccgatccc atgagctcaa tgcattttca ttcttgccga
     2581 aatagtagca gctttgagga cagcagcttg ccaactttct tgtacaaagt tggcattata
     2641 agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc
     2701 attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt
     2761 tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     2821 atatcatcat gcctcctcta gaccagccag gacagaaatg cctcgacttc gctgctaccc
     2881 aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca gtggacataa
     2941 gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga
     3001 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac
     3061 tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
     3121 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa
     3181 acaaagttaa acattatgag ggaagcggtg atcgccgaag tatcgactca actatcagag
     3241 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc
     3301 tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc
     3361 gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct
     3421 tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac
     3481 atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat
     3541 gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg
     3601 acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat
     3661 ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg
     3721 ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac
     3781 agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc
     3841 ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa
     3901 gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc
     3961 gagatcacca aggtagtcgg caaataaccc tcgagccacc catgaccaaa atcccttaac
     4021 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4081 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4141 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4201 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4261 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4321 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4381 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4441 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4501 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4561 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4621 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4681 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4741 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4801 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     4861 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     4921 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     4981 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     5041 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     5101 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     5161 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     5221 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc
     5281 agtcttaagc tcgggccc
//
