LOCUS       sequence-125748-        4994 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF clone of human BRAF [NM_004333.4] with stop codon (for 
            native or N-terminal fusions).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4994)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 4994)
  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"
COMMENT     Sequence Label: R777-E015 Hs.BRAF
FEATURES             Location/Qualifiers
     source          1..4994
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..45
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     37..54
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     38..54
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    84..183
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      184..193
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2485..2584)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2961..3752
                     /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      3845..4433
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4334..4353
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4587..4604
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4763..4790)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4882..4968)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaaa
       61 attcgagctc ggtccgagga tccgaaataa tgattttatt ttgactgata gtgacctgtt
      121 cgttgcaaca aattgataag caatgctttt ttataatgcc aactttgtac aaaaaagttg
      181 gcaccatggc ggcgctgagc ggtggcggtg gtggcggcgc ggagccgggc caggctctgt
      241 tcaacgggga catggagccc gaggccggcg ccggcgccgg cgccgcggcc tcttcggctg
      301 cggaccctgc cattccggag gaggtgtgga atatcaaaca aatgattaag ttgacacagg
      361 aacatataga ggccctattg gacaaatttg gtggggagca taatccacca tcaatatatc
      421 tggaggccta tgaagaatac accagcaagc tagatgcact ccaacaaaga gaacaacagt
      481 tattggaatc tctggggaac ggaactgatt tttctgtttc tagctctgca tcaatggata
      541 ccgttacatc ttcttcctct tctagccttt cagtgctacc ttcatctctt tcagtttttc
      601 aaaatcccac agatgtggca cggagcaacc ccaagtcacc acaaaaacct atcgttagag
      661 tcttcctgcc caacaaacag aggacagtgg tacctgcaag gtgtggagtt acagtccgag
      721 acagtctaaa gaaagcactg atgatgagag gtctaatccc agagtgctgt gctgtttaca
      781 gaattcagga tggagagaag aaaccaattg gttgggacac tgatatttcc tggcttactg
      841 gagaagaatt gcatgtggaa gtgttggaga atgttccact tacaacacac aactttgtac
      901 gaaaaacgtt tttcacctta gcattttgtg acttttgtcg aaagctgctt ttccagggtt
      961 tccgctgtca aacatgtggt tataaatttc accagcgttg tagtacagaa gttccactga
     1021 tgtgtgttaa ttatgaccaa cttgatttgc tgtttgtctc caagttcttt gaacaccacc
     1081 caataccaca ggaagaggcg tccttagcag agactgccct aacatctgga tcatcccctt
     1141 ccgcacccgc ctcggactct attgggcccc aaattctcac cagtccgtct ccttcaaaat
     1201 ccattccaat tccacagccc ttccgaccag cagatgaaga tcatcgaaat caatttgggc
     1261 aacgagaccg atcctcatca gctcccaatg tgcatataaa cacaatagaa cctgtcaata
     1321 ttgatgactt gattagagac caaggatttc gtggtgatgg aggatcaacc acaggtttgt
     1381 ctgctacccc ccctgcctca ttacctggct cactaactaa cgtgaaagcc ttacagaaat
     1441 ctccaggacc tcagcgagaa aggaagtcat cttcatcctc agaagacagg aatcgaatga
     1501 aaacacttgg tagacgggac tcgagtgatg attgggagat tcctgatggg cagattacag
     1561 tgggacaaag aattggatct ggatcatttg gaacagtcta caagggaaag tggcatggtg
     1621 atgtggcagt gaaaatgttg aatgtgacag cacctacacc tcagcagtta caagccttca
     1681 aaaatgaagt aggagtactc aggaaaacac gacatgtgaa tatcctactc ttcatgggct
     1741 attccacaaa gccacaactg gctattgtta cccagtggtg tgagggctcc agcttgtatc
     1801 accatctcca tatcattgag accaaatttg agatgatcaa acttatagat attgcacgac
     1861 agactgcaca gggcatggat tacttacacg ccaagtcaat catccacaga gacctcaaga
     1921 gtaataatat atttcttcat gaagacctca cagtaaaaat aggtgatttt ggtctagcta
     1981 cagtgaaatc tcgatggagt gggtcccatc agtttgaaca gttgtctgga tccattttgt
     2041 ggatggcacc agaagtcatc agaatgcaag ataaaaatcc atacagcttt cagtcagatg
     2101 tatatgcatt tggaattgtt ctgtatgaat tgatgactgg acagttacct tattcaaaca
     2161 tcaacaacag ggaccagata atttttatgg tgggacgagg atacctgtct ccagatctca
     2221 gtaaggtacg gagtaactgt ccaaaagcca tgaagagatt aatggcagag tgcctcaaaa
     2281 agaaaagaga tgagagacca ctctttcccc aaattctcgc ctctattgag ctgctggccc
     2341 gctcattgcc aaaaattcac cgcagtgcat cagaaccctc cttgaatcgg gctggtttcc
     2401 aaacagagga ttttagtcta tatgcttgtg cttctccaaa aacacccatc caggcagggg
     2461 gatatggtgc gtttcctgtc cactaatcaa ctttcttgta caaagttggc attataggaa
     2521 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
     2581 tttgcatgca agcttagatc ctctagacca gccaggacag aaatgcctcg acttcgctgc
     2641 tacccaaggt tgccgggtga cgcacaccgt ggaaacggat gaaggcacga acccagtgga
     2701 cataagcctg ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca cgcaactggt
     2761 ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt
     2821 atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg
     2881 ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc
     2941 ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc cgaagtatcg actcaactat
     3001 cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt
     3061 acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg
     3121 tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt
     3181 cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg
     3241 acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc
     3301 gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct
     3361 tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct
     3421 ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga
     3481 actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt
     3541 ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg
     3601 agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac
     3661 aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga
     3721 aaggcgagat caccaaggta gtcggcaaat aaccctcgag ccacccatga ccaaaatccc
     3781 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3841 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3901 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3961 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     4021 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4081 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4141 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4201 cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc acgcttcccg
     4261 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4321 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4381 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4441 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4501 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4561 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     4621 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     4681 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc
     4741 aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg
     4801 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac
     4861 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     4921 gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt
     4981 tccctactct cgcg
//
