LOCUS       sequence-424760-        5481 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 5481)
  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 5481)
  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-RPS6KA1
FEATURES             Location/Qualifiers
     source          1..5481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     287..304
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(463..490)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(582..668)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     717..739
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     731..748
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     732..748
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    764..863
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3069..3168)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3185..3204)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3186..3204)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3209..3225)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3209..3225)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3656..4447
                     /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      4540..5128
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
       61 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
      121 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
      181 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
      241 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
      301 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
      361 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
      421 taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg
      481 atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct
      541 ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga
      601 gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt
      661 tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca
      721 gtcacgacgt tgtaaaacga cggccagtct taagctcggg ccccaaataa tgattttatt
      781 ttgactgata gtgacctgtt cgttgcaaca aattgatgag caatgctttt ttataatgcc
      841 aactttgtac aaaaaagttg gcatgccgct cgcgcagctc aaggagccct ggccgctcat
      901 ggagctagtg ccgctggacc cggagaatgg acagacctca ggggaagaag ctggacttca
      961 gccgtccaag gatgagggcg tcctcaagga gatctccatc acgcaccacg tcaaggctgg
     1021 ctctgagaag gctgatccat cccatttcga gctcctcaag gttctgggcc agggatcctt
     1081 tggcaaagtc ttcctggtgc ggaaagtcac ccggcctgac agtgggcacc tgtatgctat
     1141 gaaggtgctg aagaaggcaa cgctgaaagt acgtgaccgc gtccggacca agatggagag
     1201 agacatcctg gctgatgtaa atcacccatt cgtggtgaag ctgcactatg ccttccagac
     1261 cgagggcaag ctctatctca ttctggactt cctgcgtggt ggggacctct tcacccggct
     1321 ctcaaaagag gtgatgttca cggaggagga tgtgaagttt tacctggccg agctggctct
     1381 gggcctggat cacctgcaca gcctgggtat catttacaga gacctcaagc ctgagaacat
     1441 ccttctggat gaggagggcc acatcaaact cactgacttt ggcctgagca aagaggccat
     1501 tgaccacgag aagaaggcct attctttctg cgggacagtg gagtacatgg cccctgaggt
     1561 cgtcaaccgc cagggccact cccatagtgc ggactggtgg tcctatgggg tgttgatgtt
     1621 tgagatgctg acgggctccc tgcccttcca ggggaaggac cggaaggaga ccatgacact
     1681 gattctgaag gcgaagctag gcatgcccca gtttctgagc actgaagccc agagcctctt
     1741 gcgggccctg ttcaagcgga atcctgccaa ccggctcggc tccggccctg atggggcaga
     1801 ggaaatcaag cggcatgtct tctactccac cattgactgg aataagctat accgtcgtga
     1861 gatcaagcca cccttcaagc cagcagtggc tcagcctgat gacaccttct actttgacac
     1921 cgagttcacg tcccgcacac ccaaggattc cccaggcatc ccccccagcg ctggggccca
     1981 tcagctgttc cggggcttca gcttcgtggc caccggcctg atggaagacg acggcaagcc
     2041 tcgtgccccg caggcacccc tgcactcggt ggtacagcaa ctccatggga agaacctggt
     2101 ttttagtgac ggctacgtgg taaaggagac aattggtgtg ggctcctact ctgagtgcaa
     2161 gcgctgtgtc cacaaggcca ccaacatgga gtatgctgtc aaggtcattg ataagagcaa
     2221 gcgggatcct tcagaagaga ttgagattct tctgcggtat ggccagcacc ccaacatcat
     2281 cactctgaaa gatgtgtatg atgatggcaa acacgtgtac ctggtgacag agctgatgcg
     2341 gggtggggag ctgctggaca agatcctgcg gcagaagttc ttctcagagc gggaggccag
     2401 ctttgtcctg cacaccattg gcaaaactgt ggagtatctg cactcacagg gggttgtgca
     2461 cagggacctg aagcccagca acatcctgta tgtggacgag tccgggaatc ccgagtgcct
     2521 gcgcatctgt gactttggtt ttgccaaaca gctgcgggct gagaatgggc tcctcatgac
     2581 accttgctac acagccaact ttgtggcgcc tgaggtgctg aagcgccagg gctacgatga
     2641 aggctgcgac atctggagcc tgggcattct gctgtacacc atgctggcag gatatactcc
     2701 atttgccaac ggtcccagtg acacaccaga ggaaatccta acccggatcg gcagtgggaa
     2761 gtttaccctc agtgggggaa attggaacac agtttcagag acagccaagg acctggtgtc
     2821 caagatgcta cacgtggatc cccaccagcg cctcacagct aagcaggttc tgcagcatcc
     2881 atgggtcacc cagaaagaca agcttcccca aagccagctg tcccaccagg acctacagct
     2941 tgtgaaggga gccatggctg ccacgtactc cgcactcaac agctccaagc ccacccccca
     3001 gctgaagccc atcgagtcat ccatcctggc ccagcggcga gtgaggaagt tgccatccac
     3061 caccctgtgc ccaactttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     3121 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     3181 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     3241 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctcta
     3301 gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac
     3361 accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct
     3421 gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg
     3481 tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc
     3541 tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg
     3601 gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag
     3661 ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg
     3721 ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct
     3781 gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac
     3841 gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat
     3901 tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc
     3961 agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt
     4021 cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag
     4081 cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct
     4141 atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga
     4201 tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat
     4261 cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc
     4321 cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg
     4381 cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg
     4441 caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     4501 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4561 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4621 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4681 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4741 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4801 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4861 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4921 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4981 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5041 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5101 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5161 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5221 taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc
     5281 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     5341 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     5401 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     5461 tacatcaagt gtatcatatg c
//
