LOCUS       sequence-163021-        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.H574Q
FEATURES             Location/Qualifiers
     source          1..5094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      669..678
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2973..3072)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3089..3108)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3090..3108)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3113..3129)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3113..3129)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3560..4351
                     /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      4444..5032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4933..4952
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcacc atggcggcgc tgagcggtgg cggtggtggc ggcgcggagc cgggccaggc
      721 tctgttcaac ggggacatgg agcccgaggc cggcgccggc gccggcgccg cggcctcttc
      781 ggctgcggac cctgccattc cggaggaggt gtggaatatc aaacaaatga ttaagttgac
      841 acaggaacat atagaggccc tattggacaa atttggtggg gagcataatc caccatcaat
      901 atatctggag gcctatgaag aatacaccag caagctagat gcactccaac aaagagaaca
      961 acagttattg gaatctctgg ggaacggaac tgatttttct gtttctagct ctgcatcaat
     1021 ggataccgtt acatcttctt cctcttctag cctttcagtg ctaccttcat ctctttcagt
     1081 ttttcaaaat cccacagatg tggcacggag caaccccaag tcaccacaaa aacctatcgt
     1141 tagagtcttc ctgcccaaca aacagaggac agtggtacct gcaaggtgtg gagttacagt
     1201 ccgagacagt ctaaagaaag cactgatgat gagaggtcta atcccagagt gctgtgctgt
     1261 ttacagaatt caggatggag agaagaaacc aattggttgg gacactgata tttcctggct
     1321 tactggagaa gaattgcatg tggaagtgtt ggagaatgtt ccacttacaa cacacaactt
     1381 tgtacgaaaa acgtttttca ccttagcatt ttgtgacttt tgtcgaaagc tgcttttcca
     1441 gggtttccgc tgtcaaacat gtggttataa atttcaccag cgttgtagta cagaagttcc
     1501 actgatgtgt gttaattatg accaacttga tttgctgttt gtctccaagt tctttgaaca
     1561 ccacccaata ccacaggaag aggcgtcctt agcagagact gccctaacat ctggatcatc
     1621 cccttccgca cccgcctcgg actctattgg gccccaaatt ctcaccagtc cgtctccttc
     1681 aaaatccatt ccaattccac agcccttccg accagcagat gaagatcatc gaaatcaatt
     1741 tgggcaacga gaccgatcct catcagctcc caatgtgcat ataaacacaa tagaacctgt
     1801 caatattgat gacttgatta gagaccaagg atttcgtggt gatggaggat caaccacagg
     1861 tttgtctgct accccccctg cctcattacc tggctcacta actaacgtga aagccttaca
     1921 gaaatctcca ggacctcagc gagaaaggaa gtcatcttca tcctcagaag acaggaatcg
     1981 aatgaaaaca cttggtagac gggactcgag tgatgattgg gagattcctg atgggcagat
     2041 tacagtggga caaagaattg gatctggatc atttggaaca gtctacaagg gaaagtggca
     2101 tggtgatgtg gcagtgaaaa tgttgaatgt gacagcacct acacctcagc agttacaagc
     2161 cttcaaaaat gaagtaggag tactcaggaa aacacgacat gtgaatatcc tactcttcat
     2221 gggctattcc acaaagccac aactggctat tgttacccag tggtgtgagg gctccagctt
     2281 gtatcaccat ctccatatca ttgagaccaa atttgagatg atcaaactta tagatattgc
     2341 acgacagact gcacagggca tggattactt acacgccaag tcaatcatcc aaagagacct
     2401 caagagtaat aatatatttc ttcatgaaga cctcacagta aaaataggtg attttggtct
     2461 agctacagtg aaatctcgat ggagtgggtc ccatcagttt gaacagttgt ctggatccat
     2521 tttgtggatg gcaccagaag tcatcagaat gcaagataaa aatccataca gctttcagtc
     2581 agatgtatat gcatttggaa ttgttctgta tgaattgatg actggacagt taccttattc
     2641 aaacatcaac aacagggacc agataatttt tatggtggga cgaggatacc tgtctccaga
     2701 tctcagtaag gtacggagta actgtccaaa agccatgaag agattaatgg cagagtgcct
     2761 caaaaagaaa agagatgaga gaccactctt tccccaaatt ctcgcctcta ttgagctgct
     2821 ggcccgctca ttgccaaaaa ttcaccgcag tgcatcagaa ccctccttga atcgggctgg
     2881 tttccaaaca gaggatttta gtctatatgc ttgtgcttct ccaaaaacac ccatccaggc
     2941 agggggatat ggtgcgtttc ctgtccactg attgccaact ttcttgtaca aagttggcat
     3001 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     3061 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     3121 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     3181 aaatatatca tcatgcctcc tctagaccag ccaggacaga aatgcctcga cttcgctgct
     3241 acccaaggtt gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa cccagtggac
     3301 ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac gcaactggtc
     3361 cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta
     3421 tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc
     3481 cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc
     3541 taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc
     3601 agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta
     3661 cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt
     3721 gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc
     3781 ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga
     3841 cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg
     3901 caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt
     3961 gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt
     4021 tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa
     4081 ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg
     4141 gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga
     4201 gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca
     4261 agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa
     4321 aggcgagatc accaaggtag tcggcaaata accctcgagc cacccatgac caaaatccct
     4381 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4441 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4501 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4561 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4621 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4681 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4741 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4801 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4861 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4921 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4981 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5041 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
