LOCUS       sequence-330431-        4185 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 4185)
  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 4185)
  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-GCK
FEATURES             Location/Qualifiers
     source          1..4185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1528..1627)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1644..1663)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1645..1663)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1668..1684)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1668..1684)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2115..2906
                     /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      2999..3587
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3488..3507
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3741..3758
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3917..3944)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4036..4122)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4171..4185,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4185,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagttgg catgctggac gacagagcca ggatggaggc cgccaagaag gagaaggtag
      181 agcagatcct ggcagagttc cagctgcagg aggaggacct gaagaaggtg atgagacgga
      241 tgcagaagga gatggaccgc ggcctgaggc tggagaccca tgaagaggcc agtgtgaaga
      301 tgctgcccac ctacgtgcgc tccaccccag aaggctcaga agtcggggac ttcctctccc
      361 tggacctggg tggcactaac ttcagggtga tgctggtgaa ggtgggagaa ggtgaggagg
      421 ggcagtggag cgtgaagacc aaacaccaga tgtactccat ccccgaggac gccatgaccg
      481 gcactgctga gatgctcttc gactacatct ctgagtgcat ctccgacttc ctggacaagc
      541 atcagatgaa acacaagaag ctgcccctgg gcttcacctt ctcctttcct gtgaggcacg
      601 aagacatcga taagggcatc cttctcaact ggaccaaggg cttcaaggcc tcaggagcag
      661 aagggaacaa tgtcgtgggg cttctgcgag acgctatcaa acggagaggg gactttgaaa
      721 tggatgtggt ggcaatggtg aatgacacgg tggccacgat gatctcctgc tactacgaag
      781 accatcagtg cgaggtcggc atgatcgtgg gcacgggctg caatgcctgc tacatggagg
      841 agatgcagaa tgtggagctg gtggaggggg acgagggccg catgtgcgtc aataccgagt
      901 ggggcgcctt cggggactcc ggcgagctgg acgagttcct gctggagtat gaccgcctgg
      961 tggacgagag ctctgcaaac cccggtcagc agctgtatga gaagctcata ggtggcaagt
     1021 acatgggcga gctggtgcgg cttgtgctgc tcaggctcgt ggacgaaaac ctgctcttcc
     1081 acggggaggc ctccgagcag ctgcgcacac gcggagcctt cgagacgcgc ttcgtgtcgc
     1141 aggtggagag cgacacgggc gaccgcaagc agatctacaa catcctgagc acgctggggc
     1201 tgcgaccctc gaccaccgac tgcgacatcg tgcgccgcgc ctgcgagagc gtgtctacgc
     1261 gcgctgcgca catgtgctcg gcggggctgg cgggcgtcat caaccgcatg cgcgagagcc
     1321 gcagcgagga cgtaatgcgc atcactgtgg gcgtggatgg ctccgtgtac aagctgcacc
     1381 ccagcttcaa ggagcggttc catgccagcg tgcgcaggct gacgcccagc tgcgagatca
     1441 ccttcatcga gtcggaggag ggcagtggcc ggggcgcggc cctggtctcg gcggtggcct
     1501 gtaagaaggc ctgtatgctg ggccagtgcc caactttctt gtacaaagtt ggcattataa
     1561 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca
     1621 ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt
     1681 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     1741 tatcatcatg cctcctctag accagccagg acagaaatgc ctcgacttcg ctgctaccca
     1801 aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag
     1861 cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa
     1921 ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact
     1981 gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg
     2041 gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa
     2101 caaagttaaa cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg
     2161 tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct
     2221 ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg
     2281 taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt
     2341 cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca
     2401 tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg
     2461 acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga
     2521 caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc
     2581 cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc
     2641 cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca
     2701 gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc
     2761 tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag
     2821 aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg
     2881 agatcaccaa ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg
     2941 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3001 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3061 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3121 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3181 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3241 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3301 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3361 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3421 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3481 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3541 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3601 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3661 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3721 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     3781 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     3841 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag
     3901 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct
     3961 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa
     4021 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
     4081 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
     4141 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgtt
//
