LOCUS       sequence-159437-        6108 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 6108)
  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 6108)
  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-PDGFRB
FEATURES             Location/Qualifiers
     source          1..6108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(149..176)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(268..354)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     403..425
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     417..434
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     418..434
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    450..549
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2088..2234
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(3868..3967)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3984..4003)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3985..4003)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4008..4024)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4008..4024)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4455..5246
                     /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      5339..5927
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5828..5847
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6081..6098
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
      121 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
      181 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
      241 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
      301 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
      361 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
      421 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      481 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
      541 aagttggcac catgcggctt ccgggtgcga tgccagctct ggccctcaaa ggcgagctgc
      601 tgttgctgtc tctcctgtta cttctggaac cacagatctc tcagggcctg gtcgtcacac
      661 ccccggggcc agagcttgtc ctcaatgtct ccagcacctt cgttctgacc tgctcgggtt
      721 cagctccggt ggtgtgggaa cggatgtccc aggagccccc acaggaaatg gccaaggccc
      781 aggatggcac cttctccagc gtgctcacac tgaccaacct cactgggcta gacacgggag
      841 aatacttttg cacccacaat gactcccgtg gactggagac cgatgagcgg aaacggctct
      901 acatctttgt gccagatccc accgtgggct tcctccctaa tgatgccgag gaactattca
      961 tctttctcac ggaaataact gagatcacca ttccatgccg agtaacagac ccacagctgg
     1021 tggtgacact gcacgagaag aaaggggacg ttgcactgcc tgtcccctat gatcaccaac
     1081 gtggcttttt tggtatcttt gaggacagaa gctacatctg caaaaccacc attggggaca
     1141 gggaggtgga ttctgatgcc tactatgtct acagactcca ggtgtcatcc atcaacgtct
     1201 ctgtgaacgc agtgcagact gtggtccgcc agggtgagaa catcaccctc atgtgcattg
     1261 tgatcgggaa tgaggtggtc aacttcgagt ggacataccc ccgcaaagaa agtgggcggc
     1321 tggtggagcc ggtgactgac ttcctcttgg atatgcctta ccacatccgc tccatcctgc
     1381 acatccccag tgccgagtta gaagactcgg ggacctacac ctgcaatgtg acggagagtg
     1441 tgaatgacca tcaggatgaa aaggccatca acatcaccgt ggttgagagc ggctacgtgc
     1501 ggctcctggg agaggtgggc acactacaat ttgctgagct gcatcggagc cggacactgc
     1561 aggtagtgtt cgaggcctac ccaccgccca ctgtcctgtg gttcaaagac aaccgcaccc
     1621 tgggcgactc cagcgctggc gaaatcgccc tgtccacgcg caacgtgtcg gagacccggt
     1681 atgtgtcaga gctgacactg gttcgcgtga aggtggcaga ggctggccac tacaccatgc
     1741 gggccttcca tgaggatgct gaggtccagc tctccttcca gctacagatc aatgtccctg
     1801 tccgagtgct ggagctaagt gagagccacc ctgacagtgg ggaacagaca gtccgctgtc
     1861 gtggccgggg catgccccag ccgaacatca tctggtctgc ctgcagagac ctcaaaaggt
     1921 gtccacgtga gctgccgccc acgctgctgg ggaacagttc cgaagaggag agccagctgg
     1981 agactaacgt gacgtactgg gaggaggagc aggagtttga ggtggtgagc acactgcgtc
     2041 tgcagcacgt ggatcggcca ctgtcggtgc gctgcacgct gcgcaacgct gtgggccagg
     2101 acacgcagga ggtcatcgtg gtgccacact ccttgccctt taaggtggtg gtgatctcag
     2161 ccatcctggc cctggtggtg ctcaccatca tctcccttat catcctcatc atgctttggc
     2221 agaagaagcc acgttacgag atccgatgga aggtgattga gtctgtgagc tctgacggcc
     2281 atgagtacat ctacgtggac cccatgcagc tgccctatga ctccacgtgg gagctgccgc
     2341 gggaccagct tgtgctggga cgcaccctcg gctctggggc ctttgggcag gtggtggagg
     2401 ccacggctca tggcctgagc cattctcagg ccacgatgaa agtggccgtc aagatgctta
     2461 aatccacagc ccgcagcagt gagaagcaag cccttatgtc ggagctgaag atcatgagtc
     2521 accttgggcc ccacctgaac gtggtcaacc tgttgggggc ctgcaccaaa ggaggaccca
     2581 tctatatcat cactgagtac tgccgctacg gagacctggt ggactacctg caccgcaaca
     2641 aacacacctt cctgcagcac cactccgaca agcgccgccc gcccagcgcg gagctctaca
     2701 gcaatgctct gcccgttggg ctccccctgc ccagccatgt gtccttgacc ggggagagcg
     2761 acggtggcta catggacatg agcaaggacg agtcggtgga ctatgtgccc atgctggaca
     2821 tgaaaggaga cgtcaaatat gcagacatcg agtcctccaa ctacatggcc ccttacgata
     2881 actacgttcc ctctgcccct gagaggacct gccgagcaac tttgatcaac gagtctccag
     2941 tgctaagcta catggacctc gtgggcttca gctaccaggt ggccaatggc atggagtttc
     3001 tggcctccaa gaactgcgtc cacagagacc tggcggctag gaacgtgctc atctgtgaag
     3061 gcaagctggt caagatctgt gactttggcc tggctcgaga catcatgcgg gactcgaatt
     3121 acatctccaa aggcagcacc tttttgcctt taaagtggat ggctccggag agcatcttca
     3181 acagcctcta caccaccctg agcgacgtgt ggtccttcgg gatcctgctc tgggagatct
     3241 tcaccttggg tggcacccct tacccagagc tgcccatgaa cgagcagttc tacaatgcca
     3301 tcaaacgggg ttaccgcatg gcccagcctg cccatgcctc cgacgagatc tatgagatca
     3361 tgcagaagtg ctgggaagag aagtttgaga ttcggccccc cttctcccag ctggtgctgc
     3421 ttctcgagag actgttgggc gaaggttaca aaaagaagta ccagcaggtg gatgaggagt
     3481 ttctgaggag tgaccaccca gccatccttc ggtcccaggc ccgcttgcct gggttccatg
     3541 gcctccgatc tcccctggac accagctccg tcctctatac tgccgtgcag cccaatgagg
     3601 gtgacaacga ctatatcatc cccctgcctg accccaaacc cgaggttgct gacgagggcc
     3661 cactggaggg ttcccccagc ctagccagct ccaccctgaa tgaagtcaac acctcctcaa
     3721 ccatctcctg tgacagcccc ctggagcccc aggacgaacc agagccagag ccccagcttg
     3781 agctccaggt ggagccggag ccagagctgg aacagttgcc ggattcgggg tgccctgcgc
     3841 ctcgggcgga agcagaggat agcttcctgc caactttctt gtacaaagtt ggcattataa
     3901 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca
     3961 ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt
     4021 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     4081 tatcatcatg cctcctctag accagccagg acagaaatgc ctcgacttcg ctgctaccca
     4141 aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag
     4201 cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa
     4261 ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact
     4321 gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg
     4381 gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa
     4441 caaagttaaa cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg
     4501 tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct
     4561 ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg
     4621 taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt
     4681 cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca
     4741 tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg
     4801 acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga
     4861 caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc
     4921 cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc
     4981 cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca
     5041 gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc
     5101 tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag
     5161 aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg
     5221 agatcaccaa ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg
     5281 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5341 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5401 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5461 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5521 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5581 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5641 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5701 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5761 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5821 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5881 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5941 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6001 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6061 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
