LOCUS       sequence-161650-        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.L485S
FEATURES             Location/Qualifiers
     source          1..5094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      45..54
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(55..154)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(170..187)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(170..186)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(179..201)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      250..336
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      428..455
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(614..631)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(785..1373)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(865..884)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1466..2257)
                     /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     2688..2704
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2688..2704
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2709..2728
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2709..2727
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2745..2844
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 gagcctggcc cggctccgcg ccgccaccac cgccaccgct cagcgccgcc atggtgccaa
       61 cttttttgta caaagttggc attataaaaa agcattgctc atcaatttgt tgcaacgaac
      121 aggtcactat cagtcaaaat aaaatcatta tttggggccc gagcttaaga ctggccgtcg
      181 ttttacaacg tcgtgactgg gaaaacatcc atgctagcgt taacgcgaga gtagggaact
      241 gccaggcatc aaataaaacg aaaggctcag tcggaagact gggcctttcg ttttatctgt
      301 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
      361 gtgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
      421 actaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttcctggc
      481 tagcggtacg cgtattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
      541 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
      601 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
      661 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
      721 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      781 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      841 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      901 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      961 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1021 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1081 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1141 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1201 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1261 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1321 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1381 ctttgatctt ttctacgggg tctgacgctc agtggaacga cgcgtaactc acgttaaggg
     1441 attttggtca tgggtggctc gagggttatt tgccgactac cttggtgatc tcgcctttca
     1501 cgtagtggac aaattcttcc aactgatctg cgcgcgaggc caagcgatct tcttcttgtc
     1561 caagataagc ctgtctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca
     1621 ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa
     1681 tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc
     1741 atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga
     1801 gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga
     1861 tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct
     1921 gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt
     1981 gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag
     2041 tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg
     2101 taaccagcaa atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca
     2161 aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata
     2221 gttgagtcga tacttcggcg atcaccgctt ccctcataat gtttaacttt gttttagggc
     2281 gactgccctg ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccacggcgt
     2341 aacgcgcttg ctgcttggat gcccgaggca tagactgtac cccaaaaaaa cagtcataac
     2401 aagccatgaa aaccgccact gcgccgttac caccgctgcg ttcggtcaag gttctggacc
     2461 agttgcgtga gcgcatacgc tacttgcatt acagcttacg aaccgaacag gcttatgtcc
     2521 actgggttcg tgccttcatc cgtttccacg gtgtgcgtca cccggcaacc ttgggtagca
     2581 gcgaagtcga ggcatttctg tcctggctgg tctagaggag gcatgatgat atatttttat
     2641 cttgtgcaat gtaacatcag agattttgag acacgggcca gagctgccag gaaacagcta
     2701 tgaccatgta atacgactca ctatagggga tatcagctgg atggcaaata atgattttat
     2761 tttgactgat agtgacctgt tcgttgcaac aaattgataa gcaatgcttt cttataatgc
     2821 caactttgta caagaaagtt ggcaatcagt ggacaggaaa cgcaccatat ccccctgcct
     2881 ggatgggtgt ttttggagaa gcacaagcat atagactaaa atcctctgtt tggaaaccag
     2941 cccgattcaa ggagggttct gatgcactgc ggtgaatttt tggcaatgag cgggccagca
     3001 gctcaataga ggcgagaatt tggggaaaga gtggtctctc atctcttttc tttttgaggc
     3061 actctgccat taatctcttc atggcttttg gacagttact ccgtacctta ctgagatctg
     3121 gagacaggta tcctcgtccc accataaaaa ttatctggtc cctgttgttg atgtttgaat
     3181 aaggtaactg tccagtcatc aattcataca gaacaattcc aaatgcatat acatctgact
     3241 gaaagctgta tggattttta tcttgcattc tgatgacttc tggtgccatc cacaaaatgg
     3301 atccagacaa ctgttcaaac tgatgggacc cactccatcg agatttcact gtagctagac
     3361 caaaatcacc tatttttact gtgaggtctt catgaagaaa tatattatta ctcttgaggt
     3421 ctctgtggat gattgacttg gcgtgtaagt aatccatgcc ctgtgcagtc tgtcgtgcaa
     3481 tatctataag tttgatcatc tcaaatttgg tctcaatgat atggagatgg tgatacaagc
     3541 tggagccctc acaccactgg gtaacaatag ccagttgtgg ctttgtggaa tagcccatga
     3601 agagtaggat attcacatgt cgtgttttcc tgagtactcc tacttcattt ttgaaggctt
     3661 gtaactgctg aggtgtaggt gctgtcacat tcgacatttt cactgccaca tcaccatgcc
     3721 actttccctt gtagactgtt ccaaatgatc cagatccaat tctttgtccc actgtaatct
     3781 gcccatcagg aatctcccaa tcatcactcg agtcccgtct accaagtgtt ttcattcgat
     3841 tcctgtcttc tgaggatgaa gatgacttcc tttctcgctg aggtcctgga gatttctgta
     3901 aggctttcac gttagttagt gagccaggta atgaggcagg gggggtagca gacaaacctg
     3961 tggttgatcc tccatcacca cgaaatcctt ggtctctaat caagtcatca atattgacag
     4021 gttctattgt gtttatatgc acattgggag ctgatgagga tcggtctcgt tgcccaaatt
     4081 gatttcgatg atcttcatct gctggtcgga agggctgtgg aattggaatg gattttgaag
     4141 gagacggact ggtgagaatt tggggcccaa tagagtccga ggcgggtgcg gaaggggatg
     4201 atccagatgt tagggcagtc tctgctaagg acgcctcttc ctgtggtatt gggtggtgtt
     4261 caaagaactt ggagacaaac agcaaatcaa gttggtcata attaacacac atcagtggaa
     4321 cttctgtact acaacgctgg tgaaatttat aaccacatgt ttgacagcgg aaaccctgga
     4381 aaagcagctt tcgacaaaag tcacaaaatg ctaaggtgaa aaacgttttt cgtacaaagt
     4441 tgtgtgttgt aagtggaaca ttctccaaca cttccacatg caattcttct ccagtaagcc
     4501 aggaaatatc agtgtcccaa ccaattggtt tcttctctcc atcctgaatt ctgtaaacag
     4561 cacagcactc tgggattaga cctctcatca tcagtgcttt ctttagactg tctcggactg
     4621 taactccaca ccttgcaggt accactgtcc tctgtttgtt gggcaggaag actctaacga
     4681 taggtttttg tggtgacttg gggttgctcc gtgccacatc tgtgggattt tgaaaaactg
     4741 aaagagatga aggtagcact gaaaggctag aagaggaaga agatgtaacg gtatccattg
     4801 atgcagagct agaaacagaa aaatcagttc cgttccccag agattccaat aactgttgtt
     4861 ctctttgttg gagtgcatct agcttgctgg tgtattcttc ataggcctcc agatatattg
     4921 atggtggatt atgctcccca ccaaatttgt ccaatagggc ctctatatgt tcctgtgtca
     4981 acttaatcat ttgtttgata ttccacacct cctccggaat ggcagggtcc gcagccgaag
     5041 aggccgcggc gccggcgccg gcgccggcct cgggctccat gtccccgttg aaca
//
