LOCUS       sequence-323564-        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.G466A
FEATURES             Location/Qualifiers
     source          1..5094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      43..631
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     532..551
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     785..802
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(961..988)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1080..1166)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1215..1237
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1229..1246
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1230..1246
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1262..1361
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1362..1371
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3666..3765)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3782..3801)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3783..3801)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3806..3822)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3806..3822)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4253..5044
                     /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 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
       61 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
      121 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
      181 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
      241 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
      301 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
      361 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
      421 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
      481 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
      541 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
      601 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
      661 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
      721 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
      781 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
      841 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      901 cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca
      961 aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg
     1021 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
     1081 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     1141 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct
     1201 agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc
     1261 ccaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgatgagca
     1321 atgctttttt ataatgccaa ctttgtacaa aaaagttggc accatggcgg cgctgagcgg
     1381 tggcggtggt ggcggcgcgg agccgggcca ggctctgttc aacggggaca tggagcccga
     1441 ggccggcgcc ggcgccggcg ccgcggcctc ttcggctgcg gaccctgcca ttccggagga
     1501 ggtgtggaat atcaaacaaa tgattaagtt gacacaggaa catatagagg ccctattgga
     1561 caaatttggt ggggagcata atccaccatc aatatatctg gaggcctatg aagaatacac
     1621 cagcaagcta gatgcactcc aacaaagaga acaacagtta ttggaatctc tggggaacgg
     1681 aactgatttt tctgtttcta gctctgcatc aatggatacc gttacatctt cttcctcttc
     1741 tagcctttca gtgctacctt catctctttc agtttttcaa aatcccacag atgtggcacg
     1801 gagcaacccc aagtcaccac aaaaacctat cgttagagtc ttcctgccca acaaacagag
     1861 gacagtggta cctgcaaggt gtggagttac agtccgagac agtctaaaga aagcactgat
     1921 gatgagaggt ctaatcccag agtgctgtgc tgtttacaga attcaggatg gagagaagaa
     1981 accaattggt tgggacactg atatttcctg gcttactgga gaagaattgc atgtggaagt
     2041 gttggagaat gttccactta caacacacaa ctttgtacga aaaacgtttt tcaccttagc
     2101 attttgtgac ttttgtcgaa agctgctttt ccagggtttc cgctgtcaaa catgtggtta
     2161 taaatttcac cagcgttgta gtacagaagt tccactgatg tgtgttaatt atgaccaact
     2221 tgatttgctg tttgtctcca agttctttga acaccaccca ataccacagg aagaggcgtc
     2281 cttagcagag actgccctaa catctggatc atccccttcc gcacccgcct cggactctat
     2341 tgggccccaa attctcacca gtccgtctcc ttcaaaatcc attccaattc cacagccctt
     2401 ccgaccagca gatgaagatc atcgaaatca atttgggcaa cgagaccgat cctcatcagc
     2461 tcccaatgtg catataaaca caatagaacc tgtcaatatt gatgacttga ttagagacca
     2521 aggatttcgt ggtgatggag gatcaaccac aggtttgtct gctacccccc ctgcctcatt
     2581 acctggctca ctaactaacg tgaaagcctt acagaaatct ccaggacctc agcgagaaag
     2641 gaagtcatct tcatcctcag aagacaggaa tcgaatgaaa acacttggta gacgggactc
     2701 gagtgatgat tgggagattc ctgatgggca gattacagtg ggacaaagaa ttggatctgc
     2761 atcatttgga acagtctaca agggaaagtg gcatggtgat gtggcagtga aaatgttgaa
     2821 tgtgacagca cctacacctc agcagttaca agccttcaaa aatgaagtag gagtactcag
     2881 gaaaacacga catgtgaata tcctactctt catgggctat tccacaaagc cacaactggc
     2941 tattgttacc cagtggtgtg agggctccag cttgtatcac catctccata tcattgagac
     3001 caaatttgag atgatcaaac ttatagatat tgcacgacag actgcacagg gcatggatta
     3061 cttacacgcc aagtcaatca tccacagaga cctcaagagt aataatatat ttcttcatga
     3121 agacctcaca gtaaaaatag gtgattttgg tctagctaca gtgaaatctc gatggagtgg
     3181 gtcccatcag tttgaacagt tgtctggatc cattttgtgg atggcaccag aagtcatcag
     3241 aatgcaagat aaaaatccat acagctttca gtcagatgta tatgcatttg gaattgttct
     3301 gtatgaattg atgactggac agttacctta ttcaaacatc aacaacaggg accagataat
     3361 ttttatggtg ggacgaggat acctgtctcc agatctcagt aaggtacgga gtaactgtcc
     3421 aaaagccatg aagagattaa tggcagagtg cctcaaaaag aaaagagatg agagaccact
     3481 ctttccccaa attctcgcct ctattgagct gctggcccgc tcattgccaa aaattcaccg
     3541 cagtgcatca gaaccctcct tgaatcgggc tggtttccaa acagaggatt ttagtctata
     3601 tgcttgtgct tctccaaaaa cacccatcca ggcaggggga tatggtgcgt ttcctgtcca
     3661 ctgattgcca actttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     3721 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     3781 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     3841 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc tcctctagac
     3901 cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc
     3961 gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta
     4021 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
     4081 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat
     4141 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
     4201 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga
     4261 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
     4321 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
     4381 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
     4441 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
     4501 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
     4561 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
     4621 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
     4681 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
     4741 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
     4801 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
     4861 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     4921 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     4981 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa
     5041 ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgtt
//
