LOCUS       sequence-370152-        5250 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 5250)
  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 5250)
  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-FGFR1
FEATURES             Location/Qualifiers
     source          1..5250
                     /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"
     CDS             complement(514..531)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(2593..2692)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2709..2728)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2710..2728)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2733..2749)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2733..2749)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3180..3971
                     /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      4064..4652
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4553..4572
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4806..4823
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4982..5009)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5101..5187)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(5236..5250,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5250,1..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 caccatgtgg agctggaagt gcctcctctt ctgggctgtg ctggtcacag
      181 ccacactctg caccgctagg ccgtccccga ccttgcctga acaagcccag ccctggggag
      241 cccctgtgga agtggagtcc ttcctggtcc accccggtga cctgctgcag cttcgctgtc
      301 ggctgcggga cgatgtgcag agcatcaact ggctgcggga cggggtgcag ctggcggaaa
      361 gcaaccgcac ccgcatcaca ggggaggagg tggaggtgca ggactccgtg cccgcagact
      421 ccggcctcta tgcttgcgta accagcagcc cctcgggcag tgacaccacc tacttctccg
      481 tcaatgtttc agatgctctc ccctcctcgg aggatgatga tgatgatgat gactcctctt
      541 cagaggagaa agaaacagat aacaccaaac caaaccccgt agctccatat tggacatccc
      601 cagaaaagat ggaaaagaaa ttgcatgcag tgccggctgc caagacagtg aagttcaaat
      661 gcccttccag tgggacccca aaccccacac tgcgctggtt gaaaaatggc aaagaattca
      721 aacctgacca cagaattgga ggctacaagg tccgttatgc caccgggagc atcataatgg
      781 actctgtggt gccctctgac aagggcaact acacctgcat tgtggagaat gagtacggca
      841 gcatcaacca cacataccag ctggatgtcg tggagcggtc ccctcaccgg cccatcctgc
      901 aagcagggtt gcccgccaac aaaacagtgg ccctgggtag caacgtggag ttcatgtgta
      961 aggtgtacag tgacccgcag ccgcacatcc agtggctaaa gcacatcgag gtgaatggga
     1021 gcaagattgg cccagacaac ctgccttatg tccagatctt gaagactgct ggagttaata
     1081 ccaccgacaa agagatggag gtgcttcact taagaaatgt ctcctttgag gacgcagggg
     1141 agtatacgtg cttggcgggt aactctatcg gactctccca tcactttgca tggttgaccg
     1201 ttctggaagc cctggaagag aggccggcag tgatgacctc gcccctgtac ctggagatca
     1261 tcatctattg cacaggggcc ttcctcatct cctgcatggt ggggtcggtc atcgtctaca
     1321 agatgaagag tggtaccaag aagagtgact tccacagcca gatggctgtg cacaagctgg
     1381 ccaagagcat ccctctgcgc agacaggtaa cagtgtctgc tgactccagt gcatccatga
     1441 actctggggt tcttctggtt cggccatcac ggctctcctc cagtgggact cccatgctag
     1501 caggggtctc tgagtatgag cttcccgaag accctcgctg ggagctgcct cgggacagac
     1561 tggtcttagg caaacccctg ggagagggct gctttgggca ggtggtgttg gcagaggcta
     1621 tcgggctgga caaggacaaa cccaaccgtg tgaccaaagt ggctgtgaag atgttgaagt
     1681 cggacgcaac agagaaagac ttgtcagacc tgatctcaga aatggagatg atgaagatga
     1741 tcgggaagca taagaatatc atcaacctgc tgggggcctg cacgcaggat ggtcccttgt
     1801 atgtcatcgt ggagtatgcc tccaagggca acctgcggga gtacctgcag gcccggaggc
     1861 ccccagggct ggaatactgc tacaacccca gccacaaccc agaggagcag ctctcctcca
     1921 aggacctggt gtcctgcgcc taccaggtgg cccgaggcat ggagtatctg gcctccaaga
     1981 agtgcataca ccgagacctg gcagccagga atgtcctggt gacagaggac aatgtgatga
     2041 agatagcaga ctttggcctc gcacgggaca ttcaccacat cgactactat aaaaagacaa
     2101 ccaacggccg actgcctgtg aagtggatgg cacccgaggc attatttgac cggatctaca
     2161 cccaccagag tgatgtgtgg tctttcgggg tgctcctgtg ggagatcttc actctgggcg
     2221 gctccccata ccccggtgtg cctgtggagg aacttttcaa gctgctgaag gagggtcacc
     2281 gcatggacaa gcccagtaac tgcaccaacg agctgtacat gatgatgcgg gactgctggc
     2341 atgcagtgcc ctcacagaga cccaccttca agcagctggt ggaagacctg gaccgcatcg
     2401 tggccttgac ctccaaccag gagtacctgg acctgtccat gcccctggac cagtactccc
     2461 ccagctttcc cgacacccgg agctctacgt gctcctcagg ggaggattcc gtcttctctc
     2521 atgagccgct gcccgaggag ccctgcctgc cccgacaccc agcccagctt gccaatggcg
     2581 gactcaaacg ccgcccaact ttcttgtaca aagttggcat tataagaaag cattgcttat
     2641 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag
     2701 ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc
     2761 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgcctcc
     2821 tctagaccag ccaggacaga aatgcctcga cttcgctgct acccaaggtt gccgggtgac
     2881 gcacaccgtg gaaacggatg aaggcacgaa cccagtggac ataagcctgt tcggttcgta
     2941 agctgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca
     3001 gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttggggtac
     3061 agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt
     3121 tatggagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatta
     3181 tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg
     3241 agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg
     3301 gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa
     3361 caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg
     3421 agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt
     3481 atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta
     3541 tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac
     3601 atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg
     3661 atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg
     3721 gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca
     3781 aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc
     3841 agcccgtcat acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct
     3901 cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag
     3961 tcggcaaata accctcgagc cacccatgac caaaatccct taacgtgagt tacgcgtcgt
     4021 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4081 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4141 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4201 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4261 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4321 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4381 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4441 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4501 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4561 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4621 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4681 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4741 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4801 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     4861 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     4921 gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc
     4981 aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg
     5041 gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg
     5101 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
     5161 ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc
     5221 tagcatggat gttttcccag tcacgacgtt
//
