LOCUS       sequence-421914-        4650 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 4650)
  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 4650)
  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-ITK
FEATURES             Location/Qualifiers
     source          1..4650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(116..143)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(235..321)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     370..392
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     384..401
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     385..401
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    417..516
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2377..2476)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2493..2512)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2494..2512)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2517..2533)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2517..2533)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2964..3755
                     /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      3848..4436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4337..4356
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4590..4607
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
       61 agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag
      121 gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc
      181 tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt
      241 cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga
      301 ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta acgctagcat
      361 ggatgttttc ccagtcacga cgttgtaaaa cgacggccag tcttaagctc gggccccaaa
      421 taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat gagcaatgct
      481 tttttataat gccaactttg tacaaaaaag ttggcaccat gaacaacttt atcctcctgg
      541 aagaacagct catcaagaaa tcccaacaaa agagaagaac ttctccctcg aactttaaag
      601 tccgcttctt tgtgttaacc aaagccagcc tggcatactt tgaagatcgt catgggaaga
      661 agcgcacgct gaaggggtcc attgagctct cccgaatcaa atgtgttgag attgtgaaaa
      721 gtgacatcag catcccatgc cactataaat acccgtttca ggtggtgcat gacaactacc
      781 tcctatatgt gtttgctcca gatcgtgaga gccggcagcg ctgggtgctg gcccttaaag
      841 aagaaacgag gaataataac agtttggtgc ctaaatatca tcctaatttc tggatggatg
      901 ggaagtggag gtgctgttct cagctggaga agcttgcaac aggctgtgcc caatatgatc
      961 caaccaagaa tgcttcaaag aagcctcttc ctcctactcc tgaagacaac aggcgaccac
     1021 tttgggaacc tgaagaaact gtggtcattg ccttatatga ctaccaaacc aatgatcctc
     1081 aggaactcgc actgcggcgc aacgaagagt actgcctgct ggacagttct gagattcact
     1141 ggtggagagt ccaggacagg aatgggcatg aaggatatgt accaagcagt tatctggtgg
     1201 aaaaatctcc aaataatctg gaaacctatg agtggtacaa taagagtatc agccgagaca
     1261 aagctgaaaa acttcttttg gacacaggca aagaaggagc cttcatggta agggattcca
     1321 ggactgcagg aacatacacc gtgtctgttt tcaccaaggc tgttgtaagt gagaacaatc
     1381 cctgtataaa gcattatcac atcaaggaaa caaatgacaa tcctaagcga tactatgtgg
     1441 ctgaaaagta tgtgttcgat tccatccctc ttctcatcaa ctatcaccaa cataatggag
     1501 gaggcctggt gactcgactc cggtatccag tttgttttgg gaggcagaaa gccccagtta
     1561 cagcagggct gagatacggg aaatgggtga tcgacccctc agagctcact tttgtccaag
     1621 agattggcag tgggcaattt gggttggtgc atctgggcta ctggctcaac aaggacaagg
     1681 tggctatcaa aaccattcgg gaaggggcta tgtcagaaga ggacttcata gaggaggctg
     1741 aagtaatgat gaaactctct catcccaaac tggtgcagct gtatggggtg tgcctggagc
     1801 aggcccccat ctgcctggtg tttgagttca tggagcacgg ctgcctgtca gattatctac
     1861 gcacccagcg gggacttttt gctgcagaga ccctgctggg catgtgtctg gatgtgtgtg
     1921 agggcatggc ctacctggaa gaggcatgtg tcatccacag agacttggct gccagaaatt
     1981 gtttggtggg agaaaaccaa gtcatcaagg tgtctgactt tgggatgaca aggttcgttc
     2041 tggatgatca gtacaccagt tccacaggca ccaaattccc ggtgaagtgg gcatccccag
     2101 aggttttctc tttcagtcgc tatagcagca agtccgatgt gtggtcattt ggtgtgctga
     2161 tgtgggaagt tttcagtgaa ggcaaaatcc cgtatgaaaa ccgaagcaac tcagaggtgg
     2221 tggaagacat cagtaccgga tttcggttgt acaagccccg gctggcctcc acacacgtct
     2281 accagattat gaatcactgc tggaaagaga gaccagaaga tcggccagcc ttctccagac
     2341 tgctgcgtca actggctgaa attgcagaat caggacttcc aactttcttg tacaaagttg
     2401 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     2461 ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca
     2521 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     2581 ataaaaatat atcatcatgc ctcctctaga ccagccagga cagaaatgcc tcgacttcgc
     2641 tgctacccaa ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt
     2701 ggacataagc ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact
     2761 ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt
     2821 gttatgactg tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt
     2881 acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc
     2941 gccctaaaac aaagttaaac attatgaggg aagcggtgat cgccgaagta tcgactcaac
     3001 tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt
     3061 tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta
     3121 cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa
     3181 cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc
     3241 acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc
     3301 agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta
     3361 tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac
     3421 tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat
     3481 ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca
     3541 tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa
     3601 tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg
     3661 gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg
     3721 tgaaaggcga gatcaccaag gtagtcggca aataaccctc gagccaccca tgaccaaaat
     3781 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3841 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3901 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3961 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4021 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4081 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4141 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4201 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4261 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4321 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4381 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4441 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4501 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4561 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4621 gcccaatacg caaaccgcct ctccccgcgc
//
