LOCUS       sequence-343696-        5130 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 5130)
  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 5130)
  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-PFKM
FEATURES             Location/Qualifiers
     source          1..5130
                     /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(2441..2540)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2557..2576)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2558..2576)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2581..2597)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2581..2597)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3028..3819
                     /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      3912..4500
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4401..4420
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4654..4671
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4830..4857)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4949..5035)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5084..5106
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5098..5115
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5099..5115
                     /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 tgacccatga agagcaccat
      121 gcagccaaaa ccctggggat tggcaaagcc attgctgtct taacctctgg tggagatgcc
      181 caaggtatga atgctgctgt cagggctgtg gttcgagttg gtatcttcac cggtgcccgt
      241 gtcttctttg tccatgaggg ttatcaaggc ctggtggatg gtggagatca catcaaggaa
      301 gccacctggg agagcgtttc gatgatgctt cagctgggag gcacggtgat tggaagtgcc
      361 cggtgcaagg actttcggga acgagaagga cgactccgag ctgcctacaa cctggtgaag
      421 cgtgggatca ccaatctctg tgtcattggg ggtgatggca gcctcactgg ggctgacacc
      481 ttccgttctg agtggagtga cttgttgagt gacctccaga aagcaggtaa gatcacagat
      541 gaggaggcta cgaagtccag ctacctgaac attgtgggcc tggttgggtc aattgacaat
      601 gacttctgtg gcaccgatat gaccattggc actgactctg ccctgcatcg gatcatggaa
      661 attgtagatg ccatcactac cactgcccag agccaccaga ggacatttgt gttagaagta
      721 atgggccgcc actgtggata cctggccctt gtcacctctc tgtcctgtgg ggccgactgg
      781 gtttttattc ctgaatgtcc accagatgac gactgggagg aacacctttg tcgccgactc
      841 agcgagacaa ggacccgtgg ttctcgtctc aacatcatca ttgtggctga gggtgcaatt
      901 gacaagaatg gaaaaccaat cacctcagaa gacatcaaga atctggtggt taagcgtctg
      961 ggatatgaca cccgggttac tgtcttgggg catgtgcaga ggggtgggac gccatcagcc
     1021 tttgacagaa ttctgggcag caggatgggt gtggaagcag tgatggcact tttggagggg
     1081 accccagata ccccagcctg tgtagtgagc ctctctggta accaggctgt gcgcctgccc
     1141 ctcatggaat gtgtccaggt gaccaaagat gtgaccaagg ccatggatga gaagaaattt
     1201 gacgaagccc tgaagctgag aggccggagc ttcatgaaca actgggaggt gtacaagctt
     1261 ctagctcatg tcagaccccc ggtatctaag agtggttcgc acacagtggc tgtgatgaac
     1321 gtgggggctc cggctgcagg catgaatgct gctgttcgct ccactgtgag gattggcctt
     1381 atccagggca accgagtgct cgttgtccat gatggtttcg agggcctggc caaggggcag
     1441 atagaggaag ctggctggag ctatgttggg ggctggactg gccaaggtgg ctctaaactt
     1501 gggactaaaa ggactctacc caagaagagc tttgaacaga tcagtgccaa tataactaag
     1561 tttaacattc agggccttgt catcattggg ggctttgagg cttacacagg gggcctggaa
     1621 ctgatggagg gcaggaagca gtttgatgag ctctgcatcc catttgtggt cattcctgct
     1681 acagtctcca acaatgtccc tggctcagac ttcagcgttg gggctgacac agcactcaat
     1741 actatctgca caacctgtga ccgcatcaag cagtcagcag ctggcaccaa gcgtcgggtg
     1801 tttatcattg agactatggg tggctactgt ggctacctgg ctaccatggc tggactggca
     1861 gctggggccg atgctgccta catttttgag gagcccttca ccattcgaga cctgcaggca
     1921 aatgttgaac atctggtgca aaagatgaaa acaactgtga aaaggggctt ggtgttaagg
     1981 aatgaaaagt gcaatgagaa ctataccact gacttcattt tcaacctgta ctctgaggag
     2041 gggaagggca tcttcgacag caggaagaat gtgcttggtc acatgcagca gggtgggagc
     2101 ccaaccccat ttgataggaa ttttgccact aagatgggcg ccaaggctat gaactggatg
     2161 tctgggaaaa tcaaagagag ttaccgtaat gggcggatct ttgccaatac tccagattcg
     2221 ggctgtgttc tggggatgcg taagagggct ctggtcttcc aaccagtggc tgagctgaag
     2281 gaccagacag attttgagca tcgaatcccc aaggaacagt ggtggctgaa actgaggccc
     2341 atcctcaaaa tcctagccaa gtacgagatt gacttggaca cttcagacca tgcccacctg
     2401 gagcacatca cccggaagcg gtccggggaa gctgccgtct acccaacttt cttgtacaaa
     2461 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     2521 caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg
     2581 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     2641 caagataaaa atatatcatc atgcctcctc tagaccagcc aggacagaaa tgcctcgact
     2701 tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc
     2761 cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc
     2821 aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg
     2881 gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg
     2941 cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc
     3001 agtcgcccta aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact
     3061 caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta
     3121 catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg
     3181 gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg
     3241 gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt
     3301 gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa
     3361 tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg
     3421 gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag
     3481 gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg
     3541 ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc
     3601 cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg
     3661 gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat
     3721 cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac
     3781 tacgtgaaag gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca
     3841 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     3901 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3961 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4021 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4081 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4141 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4201 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4261 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4321 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4381 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4441 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4501 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4561 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     4621 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     4681 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     4741 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
     4801 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     4861 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
     4921 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
     4981 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
     5041 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt
     5101 aaaacgacgg ccagtcttaa gctcgggccc
//
