LOCUS       sequence-257404-        4953 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 4953)
  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 4953)
  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-MAPK6
FEATURES             Location/Qualifiers
     source          1..4953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2832..2931)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2948..2967)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2949..2967)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2972..2988)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2972..2988)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3419..4210
                     /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      4303..4891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4792..4811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcatg gcagagaaat ttgaaagtct catgaacatt catggttttg atctgggttc
      721 taggtatatg gacttaaaac cattgggttg tggaggcaat ggcttggttt tttctgctgt
      781 agacaatgac tgtgacaaaa gagtagccat caagaaaatt gtccttactg atccccagag
      841 tgtcaaacat gctctacgtg aaatcaaaat tattagaaga cttgaccatg ataacattgt
      901 gaaagtgttt gagattcttg gtcccagtgg aagccaatta acagacgatg tgggctctct
      961 tacggaactg aacagtgttt acattgttca ggagtacatg gagacagact tggctaatgt
     1021 gctggagcag ggccctttac tggaagagca tgccaggctt ttcatgtatc agctgctacg
     1081 ggggctcaag tatattcact ctgcaaatgt actgcacaga gatctcaaac cagctaatct
     1141 tttcattaat acggaagact tggtgctgaa gataggtgac tttggtcttg cacggatcat
     1201 ggatcctcat tattcccata agggtcatct ttctgaagga ttggttacta aatggtacag
     1261 atctccacgt cttttacttt ctcctaataa ttatactaaa gccattgaca tgtgggctgc
     1321 aggctgcatc tttgctgaaa tgctgactgg taaaaccctt tttgcaggtg cacatgaact
     1381 tgaacagatg cagctgattt tagaatctat tcctgttgta catgaggaag atcgtcagga
     1441 gcttctcagc gtaattccag tttacattag aaatgacatg actgagccac acaaaccttt
     1501 aactcagctg cttccaggaa ttagtcgaga agcagtggat ttcctggaac aaattttgac
     1561 atttagcccc atggatcggt taacagcaga agaagcactc tcccatcctt acatgagcat
     1621 atattctttt ccaatggatg agccaatttc aagccatcct tttcatattg aagatgaagt
     1681 tgatgatatt ttgcttatgg atgaaactca cagtcacatt tataactggg aaaggtatca
     1741 tgattgtcag ttttcagagc atgattggcc tgtacataac aactttgata ttgatgaagt
     1801 tcagcttgat ccaagagctc tgtccgatgt cactgatgaa gaagaagtac aagttgatcc
     1861 ccgaaaatat ttggatggag atcgggaaaa gtatctggag gatcctgctt ttgataccaa
     1921 ttactctact gagccttgtt ggcaatactc agatcatcat gaaaacaaat attgtgatct
     1981 ggagtgtagc catacttgta actacaaaac gaggtcatca tcatatttag ataacttagt
     2041 ttggagagag agtgaagtta accattacta tgaacccaag cttattatag atctttccaa
     2101 ttggaaagaa caaagcaaag aaaaatctga taagaaaggc aaatcaaaat gtgaaaggaa
     2161 tggattggtt aaagcccaga tagcgctaga ggaagcatca cagcaactgg ctggaaaaga
     2221 aagggaaaag aatcagggat ttgattttga ttcctttatt gcaggaacta ttcagcttag
     2281 ttcccagcat gagcctactg atgttgttga taaattaaat gacttgaata gctcagtgtc
     2341 ccaactagaa ttgaaaagtt tgatatcaaa gtcagtaagc caagaaaaac aggaaaaagg
     2401 aatggcaaat ctggctcaat tagaagcctt gtaccagtct tcttgggaca gccagtttgt
     2461 gagtggtggg gaggactgtt ttttcataaa tcagttttgt gaggtaagga aggatgaaca
     2521 agttgagaag gaaaacactt acactagtta cttggacaag ttctttagca ggaaagaaga
     2581 tactgaaatg ctagaaactg agccagtaga ggatgggaag cttggggaga gaggacatga
     2641 ggaaggattt ctgaacaaca gtggggagtt cctctttaac aagcagctcg agtccatagg
     2701 catcccacag tttcacagtc cagttgggtc accacttaag tcaatacagg ccacattaac
     2761 accttctgct atgaaatctt cccctcaaat tcctcatcaa acatacagca gcattctgaa
     2821 acatctgaac tacccaactt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2881 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2941 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     3001 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgcctcct
     3061 ctagaccagc caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg
     3121 cacaccgtgg aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa
     3181 gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag
     3241 cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca
     3301 gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt
     3361 atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat
     3421 gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga
     3481 gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg
     3541 cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac
     3601 aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga
     3661 gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta
     3721 tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat
     3781 cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca
     3841 tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga
     3901 tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg
     3961 cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa
     4021 aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca
     4081 gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc
     4141 gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt
     4201 cggcaaataa ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     4261 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4321 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4381 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4441 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4501 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4561 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4621 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4681 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4741 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4801 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4861 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4921 cctggccttt tgctggcctt ttgctcacat gtt
//
