LOCUS       sequence-427166-        5102 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway cloning donor vector containing BRAF ASD mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5102)
  AUTHORS   Allan, Loewen Labs
  TITLE     Allan and Loewen Labs SFARI funded autism variants
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5102)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: BRAF - R146W in pENTR
FEATURES             Location/Qualifiers
     source          1..5102
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     47..69
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     61..78
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     62..78
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    94..193
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      200..209
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2506..2605)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2622..2641)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2623..2641)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2646..2662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2646..2662)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3093..3884
                     /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      3977..4565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4466..4485
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4719..4736
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4895..4922)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5014..5100)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt
       61 tgtaaaacga cggccagtct taagctcggg ccccaaataa tgattttatt ttgactgata
      121 gtgacctgtt cgttgcaaca aattgatgag caatgctttt ttataatgcc aactttgtac
      181 aaaaaagcag gctccggcac catggcggcg ctgagcggtg gcggtggtgg cggcgcggag
      241 ccgggccagg ctctgttcaa cggggacatg gagcccgagg ccggcgccgg cgccggcgcc
      301 gcggcctctt cggctgcgga ccctgccatt ccggaggagg tgtggaatat caaacaaatg
      361 attaagttga cacaggaaca tatagaggcc ctattggaca aatttggtgg ggagcataat
      421 ccaccatcaa tatatctgga ggcctatgaa gaatacacca gcaagctaga tgcactccaa
      481 caaagagaac aacagttatt ggaatctctg gggaacggaa ctgatttttc tgtttctagc
      541 tctgcatcaa tggataccgt tacatcttct tcctcttcta gcctttcagt gctaccttca
      601 tctctttcag tttttcaaaa tcccacagat gtggcatgga gcaaccccaa gtcaccacaa
      661 aaacctatcg ttagagtctt cctgcccaac aaacagagga cagtggtacc tgcaaggtgt
      721 ggagttacag tccgagacag tctaaagaaa gcactgatga tgagaggtct aatcccagag
      781 tgctgtgctg tttacagaat tcaggatgga gagaagaaac caattggttg ggacactgat
      841 atttcctggc ttactggaga agaattgcat gtggaagtgt tggagaatgt tccacttaca
      901 acacacaact ttgtacgaaa aacgtttttc accttagcat tttgtgactt ttgtcgaaag
      961 ctgcttttcc agggtttccg ctgtcaaaca tgtggttata aatttcacca gcgttgtagt
     1021 acagaagttc cactgatgtg tgttaattat gaccaacttg atttgctgtt tgtctccaag
     1081 ttctttgaac accacccaat accacaggaa gaggcgtcct tagcagagac tgccctaaca
     1141 tctggatcat ccccttccgc acccgcctcg gactctattg ggccccaaat tctcaccagt
     1201 ccgtctcctt caaaatccat tccaattcca cagcccttcc gaccagcaga tgaagatcat
     1261 cgaaatcaat ttgggcaacg agaccgatcc tcatcagctc ccaatgtgca tataaacaca
     1321 atagaacctg tcaatattga tgacttgatt agagaccaag gatttcgtgg tgatggagga
     1381 tcaaccacag gtttgtctgc taccccccct gcctcattac ctggctcact aactaacgtg
     1441 aaagccttac agaaatctcc aggacctcag cgagaaagga agtcatcttc atcctcagaa
     1501 gacaggaatc gaatgaaaac acttggtaga cgggactcga gtgatgattg ggagattcct
     1561 gatgggcaga ttacagtggg acaaagaatt ggatctggat catttggaac agtctacaag
     1621 ggaaagtggc atggtgatgt ggcagtgaaa atgttgaatg tgacagcacc tacacctcag
     1681 cagttacaag ccttcaaaaa tgaagtagga gtactcagga aaacacgaca tgtgaatatc
     1741 ctactcttca tgggctattc cacaaagcca caactggcta ttgttaccca gtggtgtgag
     1801 ggctccagct tgtatcacca tctccatatc attgagacca aatttgagat gatcaaactt
     1861 atagatattg cacgacagac tgcacagggc atggattact tacacgccaa gtcaatcatc
     1921 cacagagacc tcaagagtaa taatatattt cttcatgaag acctcacagt aaaaataggt
     1981 gattttggtc tagctacagt gaaatctcga tggagtgggt cccatcagtt tgaacagttg
     2041 tctggatcca ttttgtggat ggcaccagaa gtcatcagaa tgcaagataa aaatccatac
     2101 agctttcagt cagatgtata tgcatttgga attgttctgt atgaattgat gactggacag
     2161 ttaccttatt caaacatcaa caacagggac cagataattt ttatggtggg acgaggatac
     2221 ctgtctccag atctcagtaa ggtacggagt aactgtccaa aagccatgaa gagattaatg
     2281 gcagagtgcc tcaaaaagaa aagagatgag agaccactct ttccccaaat tctcgcctct
     2341 attgagctgc tggcccgctc attgccaaaa attcaccgca gtgcatcaga accctccttg
     2401 aatcgggctg gtttccaaac agaggatttt agtctatatg cttgtgcttc tccaaaaaca
     2461 cccatccagg cagggggata tggtgcgttt cctgtccact gatagaccca gctttcttgt
     2521 acaaagttgg cattataaaa aataattgct catcaatttg ttgcaacgaa caggtcacta
     2581 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
     2641 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
     2701 ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac agaaatgcct
     2761 cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac
     2821 gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct
     2881 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
     2941 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc
     3001 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
     3061 agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat
     3121 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg
     3181 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt
     3241 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc
     3301 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca
     3361 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg
     3421 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg
     3481 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg
     3541 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct
     3601 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt
     3661 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg
     3721 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg
     3781 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg
     3841 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat
     3901 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa
     3961 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4021 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4081 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4141 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4201 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4261 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4321 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4381 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4441 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4501 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4561 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4621 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4681 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4741 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     4801 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc
     4861 gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt
     4921 ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg
     4981 ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac
     5041 cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg
     5101 at
//
