LOCUS       sequence-370693-        4551 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 4551)
  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 4551)
  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-PRKCI
FEATURES             Location/Qualifiers
     source          1..4551
                     /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"
     protein_bind    complement(1862..1961)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1978..1997)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1979..1997)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2002..2018)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2002..2018)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2449..3240
                     /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      3333..3921
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3822..3841
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4075..4092
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4251..4278)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4370..4456)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4505..4527
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4519..4536
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4520..4536
                     /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 tgtcccacac ggtcgcaggc
      121 ggcggcagcg gggaccattc ccaccaggtc cgggtgaaag cctactaccg cggggatatc
      181 atgataacac attttgaacc ttccatctcc tttgagggcc tttgcaatga ggttcgagac
      241 atgtgttctt ttgacaacga acagctcttc accatgaaat ggatagatga ggaaggagac
      301 ccgtgtacag tatcatctca gttggagtta gaagaagcct ttagacttta tgagctaaac
      361 aaggattctg aactcttgat tcatgtgttc ccttgtgtac cagaacgtcc tgggatgcct
      421 tgtccaggag aagataaatc catctaccgt agaggtgcac gccgctggag aaagctttat
      481 tgtgccaatg gccacacttt ccaagccaag cgtttcaaca ggcgtgctca ctgtgccatc
      541 tgcacagacc gaatatgggg acttggacgc caaggatata agtgcatcaa ctgcaaactc
      601 ttggttcata agaagtgcca taaactcgtc acaattgaat gtgggcggca ttctttgcca
      661 caggaaccag tgatgcccat ggatcagtca tccatgcatt ctgaccatgc acagacagta
      721 attccatata atccttcaag tcatgagagt ttggatcaag ttggtgaaga aaaagaggca
      781 atgaacacca gggaaagtgg caaagcttca tccagtctag gtcttcagga ttttgatttg
      841 ctccgggtaa taggaagagg aagttatgcc aaagtactgt tggttcgatt aaaaaaaaca
      901 gatcgtattt atgcaatgaa agttgtgaaa aaagagcttg ttaatgatga tgaggatatt
      961 gattgggtac agacagagaa gcatgtgttt gagcaggcat ccaatcatcc tttccttgtt
     1021 gggctgcatt cttgctttca gacagaaagc agattgttct ttgttataga gtatgtaaat
     1081 ggaggagacc taatgtttca tatgcagcga caaagaaaac ttcctgaaga acatgccaga
     1141 ttttactctg cagaaatcag tctagcatta aattatcttc atgagcgagg gataatttat
     1201 agagatttga aactggacaa tgtattactg gactctgaag gccacattaa actcactgac
     1261 tacggcatgt gtaaggaagg attacggcca ggagatacaa ccagcacttt ctgtggtact
     1321 cctaattaca ttgctcctga aattttaaga ggagaagatt atggtttcag tgttgactgg
     1381 tgggctcttg gagtgctcat gtttgagatg atggcaggaa ggtctccatt tgatattgtt
     1441 gggagctccg ataaccctga ccagaacaca gaggattatc tcttccaagt tattttggaa
     1501 aaacaaattc gcataccacg ttctatgtct gtaaaagctg caagtgttct gaagagtttt
     1561 cttaataagg accctaagga acgattgggt tgtcttcctc aaacaggatt tgctgatatt
     1621 cagggacacc cgttcttccg aaatgttgat tgggatatga tggagcaaaa acaggtggta
     1681 cctcccttta aaccaaatat ttctggggaa tttggtttgg acaactttga ttctcagttt
     1741 actaatgaac gtgtccagct cactccagat gacgatgaca ttgtgaggaa gattgatcag
     1801 tctgaatttg aaggttttga gtatatcaat cctcttttga tgtctgcaga agaaagagtc
     1861 tgcccaactt tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc
     1921 aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc
     1981 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
     2041 aatctctgat gttacattgc acaagataaa aatatatcat catgcctcct ctagaccagc
     2101 caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg
     2161 aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc
     2221 aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac
     2281 ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct
     2341 cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc
     2401 aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg
     2461 gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc
     2521 gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca
     2581 cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga
     2641 gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc
     2701 gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag
     2761 cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca
     2821 gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc
     2881 ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag
     2941 gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga
     3001 aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg
     3061 aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata
     3121 cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat
     3181 cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa
     3241 ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     3301 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3361 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3421 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     3481 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3541 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3601 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3661 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3721 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3781 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3841 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3901 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3961 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4021 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4081 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     4141 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     4201 aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat
     4261 ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc
     4321 gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac
     4381 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag
     4441 cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg
     4501 ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc c
//
