LOCUS       sequence-132743-        4809 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 4809)
  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 4809)
  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-PRKCB2
FEATURES             Location/Qualifiers
     source          1..4809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(149..176)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(268..354)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     403..425
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     417..434
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     418..434
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    450..549
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2569..2668)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2685..2704)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2686..2704)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2709..2725)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2709..2725)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3156..3947
                     /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      4040..4628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4529..4548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4782..4799
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
      121 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
      181 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
      241 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
      301 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
      361 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
      421 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      481 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
      541 aagttggcat ggctgacccg gctgcggggc cgccgccgag cgagggcgag gagagcaccg
      601 tgcgcttcgc ccgcaaaggc gccctccggc agaagaacgt gcatgaggtc aagaaccaca
      661 aattcaccgc ccgcttcttc aagcagccca ccttctgcag ccactgcacc gacttcatct
      721 ggggcttcgg gaagcaggga ttccagtgcc aagtttgctg ctttgtggtg cacaagcggt
      781 gccatgaatt tgtcacattc tcctgccctg gcgctgacaa gggtccagcc tccgatgacc
      841 cccgcagcaa acacaagttt aagatccaca cgtactccag ccccacgttt tgtgaccact
      901 gtgggtcact gctgtatgga ctcatccacc aggggatgaa atgtgacacc tgcatgatga
      961 atgtgcacaa gcgctgcgtg atgaatgttc ccagcctgtg tggcacggac cacacggagc
     1021 gccgcggccg catctacatc caggcccaca tcgacaggga cgtcctcatt gtcctcgtaa
     1081 gagatgctaa aaaccttgta cctatggacc ccaatggcct gtcagatccc tacgtaaaac
     1141 tgaaactgat tcccgatccc aaaagtgaga gcaaacagaa gaccaaaacc atcaaatgct
     1201 ccctcaaccc tgagtggaat gagacattta gatttcagct gaaagaatcg gacaaagaca
     1261 gaagactgtc agtagagatt tgggattggg atttgaccag caggaatgac ttcatgggat
     1321 ctttgtcctt tgggatttct gaacttcaga aagccagtgt tgatggctgg tttaagttac
     1381 tgagccagga ggaaggcgag tacttcaatg tgcctgtgcc accagaagga agtgaggcca
     1441 atgaagaact gcggcagaaa tttgagaggg ccaagatcag tcagggaacc aaggtcccgg
     1501 aagaaaagac gaccaacact gtctccaaat ttgacaacaa tggcaacaga gaccggatga
     1561 aactgaccga ttttaacttc ctaatggtgc tggggaaagg cagctttggc aaggtcatgc
     1621 tttcagaacg aaaaggcaca gatgagctct atgctgtgaa gatcctgaag aaggacgttg
     1681 tgatccaaga tgatgacgtg gagtgcacta tggtggagaa gcgggtgttg gccctgcctg
     1741 ggaagccgcc cttcctgacc cagctccact cctgcttcca gaccatggac cgcctgtact
     1801 ttgtgatgga gtacgtgaat gggggcgacc tcatgtatca catccagcaa gtcggccggt
     1861 tcaaggagcc ccatgctgta ttttacgctg cagaaattgc catcggtctg ttcttcttac
     1921 agagtaaggg catcatttac cgtgacctaa aacttgacaa cgtgatgctc gattctgagg
     1981 gacacatcaa gattgccgat tttggcatgt gtaaggaaaa catctgggat ggggtgacaa
     2041 ccaagacatt ctgtggcact ccagactaca tcgcccccga gataattgct tatcagccct
     2101 atgggaagtc cgtggattgg tgggcatttg gagtcctgct gtatgaaatg ttggctgggc
     2161 aggcaccctt tgaaggggag gatgaagatg aactcttcca atccatcatg gaacacaacg
     2221 tagcctatcc caagtctatg tccaaggaag ctgtggccat ctgcaaaggg ctgatgacca
     2281 aacacccagg caaacgtctg ggttgtggac ctgaaggcga acgtgatatc aaagagcatg
     2341 catttttccg gtatattgat tgggagaaac ttgaacgcaa agagatccag cccccttata
     2401 agccaaaagc ttgtgggcga aatgctgaaa acttcgaccg atttttcacc cgccatccac
     2461 cagtcctaac acctcccgac caggaagtca tcaggaatat tgaccaatca gaattcgaag
     2521 gattttcctt tgttaactct gaatttttaa aacccgaagt caagagctac ccaactttct
     2581 tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca
     2641 ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt
     2701 attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt
     2761 acattgcaca agataaaaat atatcatcat gcctcctcta gaccagccag gacagaaatg
     2821 cctcgacttc gctgctaccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg
     2881 cacgaaccca gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc
     2941 gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg
     3001 ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc
     3061 agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc
     3121 agcagggcag tcgccctaaa acaaagttaa acattatgag ggaagcggtg atcgccgaag
     3181 tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc
     3241 tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg
     3301 atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg
     3361 accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca
     3421 ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat
     3481 ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca
     3541 ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag
     3601 cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa
     3661 ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta
     3721 cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg
     3781 ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac
     3841 aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat
     3901 ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataaccc tcgagccacc
     3961 catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag
     4021 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4081 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4141 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4201 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4261 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4321 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4381 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4441 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4501 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4561 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4621 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4681 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4741 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     4801 aagcggaag
//
