LOCUS       sequence-161507-        5094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing BRAF , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5094)
  AUTHORS   Berger AH, Brooks AN, Wu X, Shrestha Y, Chouinard C, Piccioni F, 
            Bagul M, Kamburov A, Imielinski M, Hogstrom L, Zhu C, Yang X, Pantel
            S, Sakai R, Watson J, Kaplan N, Campbell JD, Singh S, Root DE, 
            Narayan R, Natoli T, Lahr DL, Tirosh I, Tamayo P, Getz G, Wong B, 
            Doench J, Subramanian A, Golub TR, Meyerson M, Boehm JS
  TITLE     High-throughput Phenotyping of Lung Cancer Somatic Mutations.
  JOURNAL   Cancer Cell. 2016 Aug 8;30(2):214-28. doi: 
            10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28.
  PUBMED    27478040
REFERENCE   2  (bases 1 to 5094)
  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: 
            "10.1016/j.ccell.2016.06.022"; journalName: "Cancer Cell"; date: 
            "2016-08-8- 8"; volume: "30"; issue: "2"; pages: "214-28"
COMMENT     Sequence Label: pDONR223_BRAF_p.W450L
FEATURES             Location/Qualifiers
     source          1..5094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      626..635
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(636..735)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(751..768)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(751..767)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(760..782)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      831..917
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1009..1036
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1195..1212)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1366..1954)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1446..1465)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2047..2838)
                     /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     3269..3285
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3269..3285
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3290..3309
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3290..3308
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3326..3425
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 gtcccaacca attggtttct tctctccatc ctgaattctg taaacagcac agcactctgg
       61 gattagacct ctcatcatca gtgctttctt tagactgtct cggactgtaa ctccacacct
      121 tgcaggtacc actgtcctct gtttgttggg caggaagact ctaacgatag gtttttgtgg
      181 tgacttgggg ttgctccgtg ccacatctgt gggattttga aaaactgaaa gagatgaagg
      241 tagcactgaa aggctagaag aggaagaaga tgtaacggta tccattgatg cagagctaga
      301 aacagaaaaa tcagttccgt tccccagaga ttccaataac tgttgttctc tttgttggag
      361 tgcatctagc ttgctggtgt attcttcata ggcctccaga tatattgatg gtggattatg
      421 ctccccacca aatttgtcca atagggcctc tatatgttcc tgtgtcaact taatcatttg
      481 tttgatattc cacacctcct ccggaatggc agggtccgca gccgaagagg ccgcggcgcc
      541 ggcgccggcg ccggcctcgg gctccatgtc cccgttgaac agagcctggc ccggctccgc
      601 gccgccacca ccgccaccgc tcagcgccgc catggtgcca acttttttgt acaaagttgg
      661 cattataaaa aagcattgct catcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
      721 taaaatcatt atttggggcc cgagcttaag actggccgtc gttttacaac gtcgtgactg
      781 ggaaaacatc catgctagcg ttaacgcgag agtagggaac tgccaggcat caaataaaac
      841 gaaaggctca gtcggaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
      901 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgtgaagcaa cggcccggag
      961 ggtggcgggc aggacgcccg ccataaactg ccaggcatca aactaagcag aaggccatcc
     1021 tgacggatgg cctttttgcg tttctacaaa ctcttcctgg ctagcggtac gcgtattaat
     1081 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     1141 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     1201 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     1261 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1321 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1381 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1441 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1501 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1561 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1621 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1681 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1741 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1801 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1861 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1921 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1981 gtctgacgct cagtggaacg acgcgtaact cacgttaagg gattttggtc atgggtggct
     2041 cgagggttat ttgccgacta ccttggtgat ctcgcctttc acgtagtgga caaattcttc
     2101 caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag cctgtctagc
     2161 ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt cggcagcgac
     2221 atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca acgtaagcac
     2281 tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta aggtttcatt
     2341 tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg ccgctggacc
     2401 taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat caatgtcgat
     2461 cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc caaattgcag
     2521 ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc
     2581 tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat
     2641 caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca aatcaatatc
     2701 actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg ccagcaacgt
     2761 cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg atacttcggc
     2821 gatcaccgct tccctcataa tgtttaactt tgttttaggg cgactgccct gctgcgtaac
     2881 atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt gctgcttgga
     2941 tgcccgaggc atagactgta ccccaaaaaa acagtcataa caagccatga aaaccgccac
     3001 tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg
     3061 ctacttgcat tacagcttac gaaccgaaca ggcttatgtc cactgggttc gtgccttcat
     3121 ccgtttccac ggtgtgcgtc acccggcaac cttgggtagc agcgaagtcg aggcatttct
     3181 gtcctggctg gtctagagga ggcatgatga tatattttta tcttgtgcaa tgtaacatca
     3241 gagattttga gacacgggcc agagctgcca ggaaacagct atgaccatgt aatacgactc
     3301 actatagggg atatcagctg gatggcaaat aatgatttta ttttgactga tagtgacctg
     3361 ttcgttgcaa caaattgata agcaatgctt tcttataatg ccaactttgt acaagaaagt
     3421 tggcaatcag tggacaggaa acgcaccata tccccctgcc tggatgggtg tttttggaga
     3481 agcacaagca tatagactaa aatcctctgt ttggaaacca gcccgattca aggagggttc
     3541 tgatgcactg cggtgaattt ttggcaatga gcgggccagc agctcaatag aggcgagaat
     3601 ttggggaaag agtggtctct catctctttt ctttttgagg cactctgcca ttaatctctt
     3661 catggctttt ggacagttac tccgtacctt actgagatct ggagacaggt atcctcgtcc
     3721 caccataaaa attatctggt ccctgttgtt gatgtttgaa taaggtaact gtccagtcat
     3781 caattcatac agaacaattc caaatgcata tacatctgac tgaaagctgt atggattttt
     3841 atcttgcatt ctgatgactt ctggtgccat ccacaaaatg gatccagaca actgttcaaa
     3901 ctgatgggac ccactccatc gagatttcac tgtagctaga ccaaaatcac ctatttttac
     3961 tgtgaggtct tcatgaagaa atatattatt actcttgagg tctctgtgga tgattgactt
     4021 ggcgtgtaag taatccatgc cctgtgcagt ctgtcgtgca atatctataa gtttgatcat
     4081 ctcaaatttg gtctcaatga tatggagatg gtgatacaag ctggagccct cacaccactg
     4141 ggtaacaata gccagttgtg gctttgtgga atagcccatg aagagtagga tattcacatg
     4201 tcgtgttttc ctgagtactc ctacttcatt tttgaaggct tgtaactgct gaggtgtagg
     4261 tgctgtcaca ttcaacattt tcactgccac atcaccatgc cactttccct tgtagactgt
     4321 tccaaatgat ccagatccaa ttctttgtcc cactgtaatc tgcccatcag gaatctccaa
     4381 atcatcactc gagtcccgtc taccaagtgt tttcattcga ttcctgtctt ctgaggatga
     4441 agatgacttc ctttctcgct gaggtcctgg agatttctgt aaggctttca cgttagttag
     4501 tgagccaggt aatgaggcag ggggggtagc agacaaacct gtggttgatc ctccatcacc
     4561 acgaaatcct tggtctctaa tcaagtcatc aatattgaca ggttctattg tgtttatatg
     4621 cacattggga gctgatgagg atcggtctcg ttgcccaaat tgatttcgat gatcttcatc
     4681 tgctggtcgg aagggctgtg gaattggaat ggattttgaa ggagacggac tggtgagaat
     4741 ttggggccca atagagtccg aggcgggtgc ggaaggggat gatccagatg ttagggcagt
     4801 ctctgctaag gacgcctctt cctgtggtat tgggtggtgt tcaaagaact tggagacaaa
     4861 cagcaaatca agttggtcat aattaacaca catcagtgga acttctgtac tacaacgctg
     4921 gtgaaattta taaccacatg tttgacagcg gaaaccctgg aaaagcagct ttcgacaaaa
     4981 gtcacaaaat gctaaggtga aaaacgtttt tcgtacaaag ttgtgtgttg taagtggaac
     5041 attctccaac acttccacat gcaattcttc tccagtaagc caggaaatat cagt
//
