LOCUS       sequence-275219-        6003 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 6003)
  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 6003)
  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-PTK7
FEATURES             Location/Qualifiers
     source          1..6003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3314..3413)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3429..3446)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3429..3445)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3438..3460)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      3509..3595
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3687..3714
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3873..3890)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4044..4632)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4124..4143)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4725..5516)
                     /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"
     primer_bind     5947..5963
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5947..5963
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5968..5987
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5968..5986
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgataagcaa
       61 tgctttctta taatgccaac tttgtacaag aaagttggca ccatgggagc tgcgcgggga
      121 tccccggcca gaccccgccg gttgcctctg ctcagcgtcc tgctgctgcc gctgctgggc
      181 ggtacccaga cagccattgt cttcatcaag cagccgtcct cccaggatgc actgcagggg
      241 cgccgggcgc tgcttcgctg tgaggttgag gctccgggcc cggtacatgt gtactggctg
      301 ctcgatgggg cccctgtcca ggacacggaa cggcgtttcg cccagggcag cagcctgagc
      361 tttgcagctg tggaccggct gcaggactct ggcaccttcc agtgtgtggc tcgggatgat
      421 gtcactggag aagaagcccg cagtgccaac gcctccttca acatcaaatg gattgaggca
      481 ggtcctgtgg tcctgaagca tccagcctcg gaagctgaga tccagccaca gacccaggtc
      541 acacttcgtt gccacattga tgggcaccct cggcccacct accaatggtt ccgagatggg
      601 accccccttt ctgatggtca gagcaaccac acagtcagca gcaaggagcg gaacctgacg
      661 ctccggccag ctggtcctga gcatagtggg ctgtattcct gctgcgccca cagtgctttt
      721 ggccaggctt gcagcagcca gaacttcacc ttgagcattg ctgatgaaag ctttgccagg
      781 gtggtgctgg caccccagga cgtggtagta gcgaggtatg aggaggccat gttccattgc
      841 cagttctcag cccagccacc cccgagcctg cagtggctct ttgaggatga gactcccatc
      901 actaaccgca gtcgcccccc acacctccgc agagccacag tgtttgccaa cgggtctctg
      961 ctgctgaccc aggtccggcc acgcaatgca gggatctacc gctgcattgg ccaggggcag
     1021 aggggcccac ccatcatcct ggaagccaca cttcacctag cagagattga agacatgccg
     1081 ctatttgagc cacgggtgtt tacagctggc agcgaggagc gtgtgacctg ccttcccccc
     1141 aagggtctgc cagagcccag cgtgtggtgg gagcacgcgg gagtccggct gcccacccat
     1201 ggcagggtct accagaaggg ccacgagctg gtgttggcca atattgctga aagtgatgct
     1261 ggtgtctaca cctgccacgc ggccaacctg gctggtcagc ggagacagga tgtcaacatc
     1321 actgtggcca ctgtgccctc ctggctgaag aagccccaag acagccagct ggaggagggc
     1381 aaacccggct acttggattg cctgacccag gccacaccaa aacctacagt tgtctggtac
     1441 agaaaccaga tgctcatctc agaggactca cggttcgagg tcttcaagaa tgggaccttg
     1501 cgcatcaaca gcgtggaggt gtatgatggg acatggtacc gttgtatgag cagcacccca
     1561 gccggcagca tcgaggcgca agcccgtgtc caagtgctgg aaaagctcaa gttcacacca
     1621 ccaccccagc cacagcagtg catggagttt gacaaggagg ccacggtgcc ctgttcagcc
     1681 acaggccgag agaagcccac tattaagtgg gaacgggcag atgggagcag cctcccagag
     1741 tgggtgacag acaacgctgg gaccctgcat tttgcccggg tgactcgaga tgacgctggc
     1801 aactacactt gcattgcctc caacgggccg cagggccaga ttcgtgccca tgtccagctc
     1861 actgtggcag tttttatcac cttcaaagtg gaaccagagc gtacgactgt gtaccagggc
     1921 cacacagccc tactgcagtg cgaggcccag ggggacccca agccgctgat tcagtggaaa
     1981 ggcaaggacc gcatcctgga ccccaccaag ctgggaccca ggatgcacat cttccagaat
     2041 ggctccctgg tgatccatga cgtggcccct gaggactcag gccgctacac ctgcattgca
     2101 ggcaacagct gcaacatcaa gcacacggag gcccccctct atgtcgtgga caagcctgtg
     2161 ccggaggagt cggagggccc tggcagccct cccccctaca agatgatcca gaccattggg
     2221 ttgtcggtgg gtgccgctgt ggcctacatc attgccgtgc tgggcctcat gttctactgc
     2281 aagaagcgct gcaaagccaa gcggctgcag aagcagcccg agggcgagga gccagagatg
     2341 gaatgcctca acggtgggcc tttgcagaac gggcagccct cagcagagat ccaagaagaa
     2401 gtggccttga ccagcttggg ctccggcccc gcggccacca acaaacgcca cagcacaagt
     2461 gataagatgc acttcccacg gtctagcctg cagcccatca ccacgctggg gaagagtgag
     2521 tttggggagg tgttcctggc aaaggctcag ggcttggagg agggagtggc agagaccctg
     2581 gtacttgtga agagcctgca gagcaaggat gagcagcagc agctggactt ccggagggag
     2641 ttggagatgt ttgggaagct gaaccacgcc aacgtggtgc ggctcctggg gctgtgccgg
     2701 gaggctgagc cccactacat ggtgctggaa tatgtggatc tgggagacct caagcagttc
     2761 ctgaggattt ccaagagcaa ggatgaaaaa ttgaagtcac agcccctcag caccaagcag
     2821 aaggtggccc tatgcaccca ggtagccctg ggcatggagc acctgtccaa caaccgcttt
     2881 gtgcataagg acttggctgc gcgtaactgc ctggtcagtg cccagagaca agtgaaggtg
     2941 tctgccctgg gcctcagcaa ggatgtgtac aacagtgagt actaccactt ccgccaggcc
     3001 tgggtgccgc tgcgctggat gtcccccgag gccatcctgg agggtgactt ctctaccaag
     3061 tctgatgtct gggccttcgg tgtgctgatg tgggaagtgt ttacacatgg agagatgccc
     3121 catggtgggc aggcagatga tgaagtactg gcagatttgc aggctgggaa ggctagactt
     3181 cctcagcccg agggctgccc ttccaaactc tatcggctga tgcagcgctg ctgggccctc
     3241 agccccaagg accggccctc cttcagtgag attgccagcg ccctgggaga cagcaccgtg
     3301 gacagcaagc cgtagccaac tttcttgtac aaagttggca ttataaaaaa gcattgctca
     3361 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttggggcccg
     3421 agcttaagac tggccgtcgt tttacaacgt cgtgactggg aaaacatcca tgctagcgtt
     3481 aacgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cggaagactg
     3541 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     3601 gggagcggat ttgaacgttg tgaagcaacg gcccggaggg tggcgggcag gacgcccgcc
     3661 ataaactgcc aggcatcaaa ctaagcagaa ggccatcctg acggatggcc tttttgcgtt
     3721 tctacaaact cttcctggct agcggtacgc gtattaattg cgttgcgctc actgcccgct
     3781 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3841 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3901 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3961 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4021 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4081 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4141 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4201 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4261 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4321 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4381 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4441 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4501 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4561 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4621 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgac
     4681 gcgtaactca cgttaaggga ttttggtcat gggtggctcg agggttattt gccgactacc
     4741 ttggtgatct cgcctttcac gtagtggaca aattcttcca actgatctgc gcgcgaggcc
     4801 aagcgatctt cttcttgtcc aagataagcc tgtctagctt caagtatgac gggctgatac
     4861 tgggccggca ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc gattttgccg
     4921 gttactgcgc tgtaccaaat gcgggacaac gtaagcacta catttcgctc atcgccagcc
     4981 cagtcgggcg gcgagttcca tagcgttaag gtttcattta gcgcctcaaa tagatcctgt
     5041 tcaggaaccg gatcaaagag ttcctccgcc gctggaccta ccaaggcaac gctatgttct
     5101 cttgcttttg tcagcaagat agccagatca atgtcgatcg tggctggctc gaagatacct
     5161 gcaagaatgt cattgcgctg ccattctcca aattgcagtt cgcgcttagc tggataacgc
     5221 cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag aatctcgctc
     5281 tctccagggg aagccgaagt ttccaaaagg tcgttgatca aagctcgccg cgttgtttca
     5341 tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt caggccgcca
     5401 tccactgcgg agccgtacaa atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg
     5461 acgccaacta cctctgatag ttgagtcgat acttcggcga tcaccgcttc cctcataatg
     5521 tttaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct ccataacatc
     5581 aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtacc
     5641 ccaaaaaaac agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt
     5701 tcggtcaagg ttctggacca gttgcgtgag cgcatacgct acttgcatta cagcttacga
     5761 accgaacagg cttatgtcca ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac
     5821 ccggcaacct tgggtagcag cgaagtcgag gcatttctgt cctggctggt ctagaggagg
     5881 catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga cacgggccag
     5941 agctgccagg aaacagctat gaccatgtaa tacgactcac tataggggat atcagctgga
     6001 tgg
//
