LOCUS       sequence-167980-        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.L597Q
FEATURES             Location/Qualifiers
     source          1..5094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      18..606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     507..526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     760..777
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(936..963)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1055..1141)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1190..1212
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1204..1221
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1205..1221
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1237..1336
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1337..1346
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3641..3740)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3757..3776)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3758..3776)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3781..3797)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3781..3797)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4228..5019
                     /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 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
       61 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
      121 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
      181 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
      241 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
      301 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
      361 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
      421 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      481 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
      541 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      601 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
      661 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
      721 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
      781 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
      841 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
      901 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
      961 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     1021 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     1081 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     1141 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
     1201 cgttgtaaaa cgacggccag tcttaagctc gggccccaaa taatgatttt attttgactg
     1261 atagtgacct gttcgttgca acaaattgat gagcaatgct tttttataat gccaactttg
     1321 tacaaaaaag ttggcaccat ggcggcgctg agcggtggcg gtggtggcgg cgcggagccg
     1381 ggccaggctc tgttcaacgg ggacatggag cccgaggccg gcgccggcgc cggcgccgcg
     1441 gcctcttcgg ctgcggaccc tgccattccg gaggaggtgt ggaatatcaa acaaatgatt
     1501 aagttgacac aggaacatat agaggcccta ttggacaaat ttggtgggga gcataatcca
     1561 ccatcaatat atctggaggc ctatgaagaa tacaccagca agctagatgc actccaacaa
     1621 agagaacaac agttattgga atctctgggg aacggaactg atttttctgt ttctagctct
     1681 gcatcaatgg ataccgttac atcttcttcc tcttctagcc tttcagtgct accttcatct
     1741 ctttcagttt ttcaaaatcc cacagatgtg gcacggagca accccaagtc accacaaaaa
     1801 cctatcgtta gagtcttcct gcccaacaaa cagaggacag tggtacctgc aaggtgtgga
     1861 gttacagtcc gagacagtct aaagaaagca ctgatgatga gaggtctaat cccagagtgc
     1921 tgtgctgttt acagaattca ggatggagag aagaaaccaa ttggttggga cactgatatt
     1981 tcctggctta ctggagaaga attgcatgtg gaagtgttgg agaatgttcc acttacaaca
     2041 cacaactttg tacgaaaaac gtttttcacc ttagcatttt gtgacttttg tcgaaagctg
     2101 cttttccagg gtttccgctg tcaaacatgt ggttataaat ttcaccagcg ttgtagtaca
     2161 gaagttccac tgatgtgtgt taattatgac caacttgatt tgctgtttgt ctccaagttc
     2221 tttgaacacc acccaatacc acaggaagag gcgtccttag cagagactgc cctaacatct
     2281 ggatcatccc cttccgcacc cgcctcggac tctattgggc cccaaattct caccagtccg
     2341 tctccttcaa aatccattcc aattccacag cccttccgac cagcagatga agatcatcga
     2401 aatcaatttg ggcaacgaga ccgatcctca tcagctccca atgtgcatat aaacacaata
     2461 gaacctgtca atattgatga cttgattaga gaccaaggat ttcgtggtga tggaggatca
     2521 accacaggtt tgtctgctac cccccctgcc tcattacctg gctcactaac taacgtgaaa
     2581 gccttacaga aatctccagg acctcagcga gaaaggaagt catcttcatc ctcagaagac
     2641 aggaatcgaa tgaaaacact tggtagacgg gactcgagtg atgattggga gattcctgat
     2701 gggcagatta cagtgggaca aagaattgga tctggatcat ttggaacagt ctacaaggga
     2761 aagtggcatg gtgatgtggc agtgaaaatg ttgaatgtga cagcacctac acctcagcag
     2821 ttacaagcct tcaaaaatga agtaggagta ctcaggaaaa cacgacatgt gaatatccta
     2881 ctcttcatgg gctattccac aaagccacaa ctggctattg ttacccagtg gtgtgagggc
     2941 tccagcttgt atcaccatct ccatatcatt gagaccaaat ttgagatgat caaacttata
     3001 gatattgcac gacagactgc acagggcatg gattacttac acgccaagtc aatcatccac
     3061 agagacctca agagtaataa tatatttctt catgaagacc tcacagtaaa aataggtgat
     3121 tttggtcaag ctacagtgaa atctcgatgg agtgggtccc atcagtttga acagttgtct
     3181 ggatccattt tgtggatggc accagaagtc atcagaatgc aagataaaaa tccatacagc
     3241 tttcagtcag atgtatatgc atttggaatt gttctgtatg aattgatgac tggacagtta
     3301 ccttattcaa acatcaacaa cagggaccag ataattttta tggtgggacg aggatacctg
     3361 tctccagatc tcagtaaggt acggagtaac tgtccaaaag ccatgaagag attaatggca
     3421 gagtgcctca aaaagaaaag agatgagaga ccactctttc cccaaattct cgcctctatt
     3481 gagctgctgg cccgctcatt gccaaaaatt caccgcagtg catcagaacc ctccttgaat
     3541 cgggctggtt tccaaacaga ggattttagt ctatatgctt gtgcttctcc aaaaacaccc
     3601 atccaggcag ggggatatgg tgcgtttcct gtccactgat tgccaacttt cttgtacaaa
     3661 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     3721 caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg
     3781 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     3841 caagataaaa atatatcatc atgcctcctc tagaccagcc aggacagaaa tgcctcgact
     3901 tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc
     3961 cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc
     4021 aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg
     4081 gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg
     4141 cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc
     4201 agtcgcccta aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact
     4261 caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta
     4321 catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg
     4381 gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg
     4441 gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt
     4501 gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa
     4561 tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg
     4621 gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag
     4681 gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg
     4741 ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc
     4801 cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg
     4861 gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat
     4921 cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac
     4981 tacgtgaaag gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca
     5041 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaa
//
