LOCUS       sequence-125749-        4991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF clone of human BRAF [NM_004333.4] without stop codon 
            (for C-terminal fusions).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4991)
  TITLE     Esposito lab
REFERENCE   2  (bases 1 to 4991)
  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-E016 Hs.BRAF-nostop
FEATURES             Location/Qualifiers
     source          1..4991
                     /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(2482..2581)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2958..3749
                     /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      3842..4430
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4331..4350
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4584..4601
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4760..4787)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4879..4965)
                     /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 cactcaactt tcttgtacaa agttggcatt ataggaaagc
     2521 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
     2581 gcatgcaagc ttagatcctc tagaccagcc aggacagaaa tgcctcgact tcgctgctac
     2641 ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat
     2701 aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca
     2761 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
     2821 actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg
     2881 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta
     2941 aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag
     3001 aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg
     3061 gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga
     3121 ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg
     3181 cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg
     3241 acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca
     3301 atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc
     3361 tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg
     3421 atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact
     3481 cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt
     3541 acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc
     3601 gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag
     3661 aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag
     3721 gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta
     3781 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3841 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3901 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3961 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     4021 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4081 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4141 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4201 cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag
     4261 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4321 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4381 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4441 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4501 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4561 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     4621 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     4681 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg
     4741 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg
     4801 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt
     4861 caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
     4921 aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc
     4981 ctactctcgc g
//
