LOCUS       sequence-168669-        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   Kim E, Ilic N, Shrestha Y, Zou L, Kamburov A, Zhu C, Yang X, Lubonja
            R, Tran N, Nguyen C, Lawrence MS, Piccioni F, Bagul M, Doench JG, 
            Chouinard CR, Wu X, Hogstrom L, Natoli T, Tamayo P, Horn H, Corsello
            SM, Lage K, Root DE, Subramanian A, Golub TR, Getz G, Boehm JS, Hahn
            WC
  TITLE     Systematic Functional Interrogation of Rare Cancer Variants 
            Identifies Oncogenic Alleles.
  JOURNAL   Cancer Discov. 2016 Jul;6(7):714-26. doi: 
            10.1158/2159-8290.CD-16-0160. Epub 2016 May 4.
  PUBMED    27147599
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: 
            "10.1158/2159-8290.CD-16-0160"; journalName: "Cancer Discov"; date: 
            "2016-07"; volume: "6"; issue: "7"; pages: "714-26"
COMMENT     Sequence Label: pDONR223_BRAF_p.G466V
FEATURES             Location/Qualifiers
     source          1..5094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      23..611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     512..531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     765..782
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(941..968)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1060..1146)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1195..1217
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1209..1226
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1210..1226
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1242..1341
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1342..1351
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3646..3745)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3762..3781)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3763..3781)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3786..3802)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3786..3802)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4233..5024
                     /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"
ORIGIN
        1 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
       61 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
      121 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
      181 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
      241 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
      301 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
      361 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
      421 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
      481 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
      541 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      601 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      661 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
      721 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
      781 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
      841 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
      901 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat
      961 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
     1021 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc
     1081 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt
     1141 tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg ttttcccagt
     1201 cacgacgttg taaaacgacg gccagtctta agctcgggcc ccaaataatg attttatttt
     1261 gactgatagt gacctgttcg ttgcaacaaa ttgatgagca atgctttttt ataatgccaa
     1321 ctttgtacaa aaaagttggc accatggcgg cgctgagcgg tggcggtggt ggcggcgcgg
     1381 agccgggcca ggctctgttc aacggggaca tggagcccga ggccggcgcc ggcgccggcg
     1441 ccgcggcctc ttcggctgcg gaccctgcca ttccggagga ggtgtggaat atcaaacaaa
     1501 tgattaagtt gacacaggaa catatagagg ccctattgga caaatttggt ggggagcata
     1561 atccaccatc aatatatctg gaggcctatg aagaatacac cagcaagcta gatgcactcc
     1621 aacaaagaga acaacagtta ttggaatctc tggggaacgg aactgatttt tctgtttcta
     1681 gctctgcatc aatggatacc gttacatctt cttcctcttc tagcctttca gtgctacctt
     1741 catctctttc agtttttcaa aatcccacag atgtggcacg gagcaacccc aagtcaccac
     1801 aaaaacctat cgttagagtc ttcctgccca acaaacagag gacagtggta cctgcaaggt
     1861 gtggagttac agtccgagac agtctaaaga aagcactgat gatgagaggt ctaatcccag
     1921 agtgctgtgc tgtttacaga attcaggatg gagagaagaa accaattggt tgggacactg
     1981 atatttcctg gcttactgga gaagaattgc atgtggaagt gttggagaat gttccactta
     2041 caacacacaa ctttgtacga aaaacgtttt tcaccttagc attttgtgac ttttgtcgaa
     2101 agctgctttt ccagggtttc cgctgtcaaa catgtggtta taaatttcac cagcgttgta
     2161 gtacagaagt tccactgatg tgtgttaatt atgaccaact tgatttgctg tttgtctcca
     2221 agttctttga acaccaccca ataccacagg aagaggcgtc cttagcagag actgccctaa
     2281 catctggatc atccccttcc gcacccgcct cggactctat tgggccccaa attctcacca
     2341 gtccgtctcc ttcaaaatcc attccaattc cacagccctt ccgaccagca gatgaagatc
     2401 atcgaaatca atttgggcaa cgagaccgat cctcatcagc tcccaatgtg catataaaca
     2461 caatagaacc tgtcaatatt gatgacttga ttagagacca aggatttcgt ggtgatggag
     2521 gatcaaccac aggtttgtct gctacccccc ctgcctcatt acctggctca ctaactaacg
     2581 tgaaagcctt acagaaatct ccaggacctc agcgagaaag gaagtcatct tcatcctcag
     2641 aagacaggaa tcgaatgaaa acacttggta gacgggactc gagtgatgat tgggagattc
     2701 ctgatgggca gattacagtg ggacaaagaa ttggatctgt atcatttgga acagtctaca
     2761 agggaaagtg gcatggtgat gtggcagtga aaatgttgaa tgtgacagca cctacacctc
     2821 agcagttaca agccttcaaa aatgaagtag gagtactcag gaaaacacga catgtgaata
     2881 tcctactctt catgggctat tccacaaagc cacaactggc tattgttacc cagtggtgtg
     2941 agggctccag cttgtatcac catctccata tcattgagac caaatttgag atgatcaaac
     3001 ttatagatat tgcacgacag actgcacagg gcatggatta cttacacgcc aagtcaatca
     3061 tccacagaga cctcaagagt aataatatat ttcttcatga agacctcaca gtaaaaatag
     3121 gtgattttgg tctagctaca gtgaaatctc gatggagtgg gtcccatcag tttgaacagt
     3181 tgtctggatc cattttgtgg atggcaccag aagtcatcag aatgcaagat aaaaatccat
     3241 acagctttca gtcagatgta tatgcatttg gaattgttct gtatgaattg atgactggac
     3301 agttacctta ttcaaacatc aacaacaggg accagataat ttttatggtg ggacgaggat
     3361 acctgtctcc agatctcagt aaggtacgga gtaactgtcc aaaagccatg aagagattaa
     3421 tggcagagtg cctcaaaaag aaaagagatg agagaccact ctttccccaa attctcgcct
     3481 ctattgagct gctggcccgc tcattgccaa aaattcaccg cagtgcatca gaaccctcct
     3541 tgaatcgggc tggtttccaa acagaggatt ttagtctata tgcttgtgct tctccaaaaa
     3601 cacccatcca ggcaggggga tatggtgcgt ttcctgtcca ctgattgcca actttcttgt
     3661 acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta
     3721 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
     3781 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
     3841 ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac agaaatgcct
     3901 cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac
     3961 gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct
     4021 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
     4081 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc
     4141 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
     4201 agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat
     4261 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg
     4321 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt
     4381 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc
     4441 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca
     4501 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg
     4561 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg
     4621 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg
     4681 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct
     4741 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt
     4801 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg
     4861 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg
     4921 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg
     4981 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat
     5041 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccg
//
