LOCUS       sequence-313982-        4857 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 4857)
  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 4857)
  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-ADRBK1
FEATURES             Location/Qualifiers
     source          1..4857
                     /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"
     primer_bind     complement(2284..2303)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2285..2303)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2308..2324)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2308..2324)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2755..3546
                     /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      3639..4227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4128..4147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4381..4398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4557..4584)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4676..4762)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4811..4833
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4825..4842
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4826..4842
                     /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 tggcggacct ggaggcggtg
      121 ctggccgacg tgagctacct gatggccatg gagaagagca aggccacgcc ggccgcgcgc
      181 gccagcaaga agatactgct gcccgagccc agcatccgca gtgtcatgca gaagtacctg
      241 gaggaccggg gcgaggtgac ctttgagaag atcttttccc agaagctggg gtacctgctc
      301 ttccgagact tctgcctgaa ccacctggag gaggccaggc ccttggtgga attctatgag
      361 gagatcaaga agtacgagaa gctggagacg gaggaggagc gtgtggcccg cagccgggag
      421 atcttcgact catacatcat gaaggagctg ctggcctgct cgcatccctt ctcgaagagt
      481 gccactgagc atgtccaagg ccacctgggg aagaagcagg tgcctccgga tctcttccag
      541 ccatacatcg aagagatttg tcaaaacctc cgaggggacg tgttccagaa attcattgag
      601 agcgataagt tcacacggtt ttgccagtgg aagaatgtgg agctcaacat ccacctgacc
      661 atgaatgact tcagcgtgca tcgcatcatt gggcgcgggg gctttggcga ggtctatggg
      721 tgccggaagg ctgacacagg caagatgtac gccatgaagt gcctggacaa aaagcgcatc
      781 aagatgaagc agggggagac cctggccctg aacgagcgca tcatgctctc gctcgtcagc
      841 actggggact gcccattcat tgtctgcatg tcatacgcgt tccacacgcc agacaagctc
      901 agcttcatcc tggacctcat gaacggtggg gacctgcact accacctctc ccagcacggg
      961 gtcttctcag aggctgacat gcgcttctat gcggccgaga tcatcctggg cctggagcac
     1021 atgcacaacc gcttcgtggt ctaccgggac ctgaagccag ccaacatcct tctggacgag
     1081 catggccacg tgcggatctc ggacctgggc ctggcctgtg acttctccaa gaagaagccc
     1141 catgccagcg tgggcaccca cgggtacatg gctccggagg tcctgcagaa gggcgtggcc
     1201 tacgacagca gtgccgactg gttctctctg gggtgcatgc tcttcaagtt gctgcggggg
     1261 cacagcccct tccggcagca caagaccaaa gacaagcatg agatcgaccg catgacgctg
     1321 acgatggccg tggagctgcc cgactccttc tcccctgaac tacgctccct gctggagggg
     1381 ttgctgcaga gggatgtcaa ccggagattg ggctgcctgg gccgaggggc tcaggaggtg
     1441 aaagagagcc cctttttccg ctccctggac tggcagatgg tcttcttgca gaagtaccct
     1501 cccccgctga tccccccacg aggggaggtg aacgcggccg acgccttcga cattggctcc
     1561 ttcgatgagg aggacacaaa aggaatcaag ttactggaca gtgatcagga gctctaccgc
     1621 aacttccccc tcaccatctc ggagcggtgg cagcaggagg tggcagagac tgtcttcgac
     1681 accatcaacg ctgagacaga ccggctggag gctcgcaaga aagccaagaa caagcagctg
     1741 ggccatgagg aagactacgc cctgggcaag gactgcatca tgcatggcta catgtccaag
     1801 atgggcaacc ccttcctgac ccagtggcag cggcggtact tctacctgtt ccccaaccgc
     1861 ctcgagtggc ggggcgaggg cgaggccccg cagagcctgc tgaccatgga ggagatccag
     1921 tcggtggagg agacgcagat caaggagcgc aagtgcctgc tcctcaagat ccgcggtggg
     1981 aaacagttca ttttgcagtg cgatagcgac cctgagctgg tgcagtggaa gaaggagctg
     2041 cgcgacgcct accgcgaggc ccagcagctg gtgcagcggg tgcccaagat gaagaacaag
     2101 ccgcgctcgc ccgtggtgga gctgagcaag gtgccgctgg tccagcgcgg cagtgccaac
     2161 ggcctctgcc aaactttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt
     2221 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat
     2281 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
     2341 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg cctcctctag
     2401 accagccagg acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca
     2461 ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg
     2521 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt
     2581 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct
     2641 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
     2701 agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg
     2761 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc
     2821 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg
     2881 aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg
     2941 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt
     3001 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca
     3061 gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc
     3121 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc
     3181 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta
     3241 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat
     3301 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc
     3361 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc
     3421 gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc
     3481 gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc
     3541 aaataaccct cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
     3601 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3661 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3721 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3781 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3841 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3901 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3961 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4021 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4081 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4141 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4201 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4261 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4321 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4381 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     4441 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     4501 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
     4561 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
     4621 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
     4681 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
     4741 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca
     4801 tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggccc
//
