LOCUS       sequence-427156-        4262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway cloning donor vector containing MECP2 ASD mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4262)
  AUTHORS   Allan, Loewen Labs
  TITLE     Allan and Loewen Labs SFARI funded autism variants
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4262)
  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            "Unpublished"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: MECP2 - E282G in pENTR
FEATURES             Location/Qualifiers
     source          1..4262
                     /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"
     CDS             1297..1317
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     protein_bind    complement(1666..1765)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1782..1801)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1783..1801)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1806..1822)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1806..1822)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2253..3044
                     /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      3137..3725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3626..3645
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3879..3896
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4055..4082)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4174..4260)
                     /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 catggtagct gggatgttag ggctcaggga agaaaagtca
      241 gaagaccagg acctccaggg cctcaaggac aaacccctca agtttaaaaa ggtgaagaaa
      301 gataagaaag aagagaaaga gggcaagcat gagcccgtgc agccatcagc ccaccactct
      361 gctgagcccg cagaggcagg caaagcagag acatcagaag ggtcaggctc cgccccggct
      421 gtgccggaag cttctgcctc ccccaaacag cggcgctcca tcatccgtga ccggggaccc
      481 atgtatgatg accccaccct gcctgaaggc tggacacgga agcttaagca aaggaaatct
      541 ggccgctctg ctgggaagta tgatgtgtat ttgatcaatc cccagggaaa agcctttcgc
      601 tctaaagtgg agttgattgc gtacttcgaa aaggtaggcg acacatccct ggaccctaat
      661 gattttgact tcacggtaac tgggagaggg agcccctccc ggcgagagca gaaaccacct
      721 aagaagccca aatctcccaa agctccagga actggcagag gccggggacg ccccaaaggg
      781 agcggcacca cgagacccaa ggcggccacg tcagagggtg tgcaggtgaa aagggtcctg
      841 gagaaaagtc ctgggaagct ccttgtcaag atgccttttc aaacttcgcc agggggcaag
      901 gctgaggggg gtggggccac cacatccacc caggtcatgg tgatcaaacg ccccggcagg
      961 aagcgaaaag ctgaggccga ccctcaggcc attcccaaga aacggggccg aaagccgggg
     1021 agtgtggtgg cagccgctgc cgccggggcc aaaaagaaag ccgtgaagga gtcttctatc
     1081 cgatctgtgc aggagaccgt actccccatc aagaagcgca agacccggga gacggtcagc
     1141 atcgaggtca aggaagtggt gaagcccctg ctggtgtcca ccctcggtga gaagagcggg
     1201 aaaggactga agacctgtaa gagccctggg cggaaaagca aggagagcag ccccaagggg
     1261 cgcagcagca gcgcctcctc accccccaag aaggagcacc accaccatca ccaccactca
     1321 gagtccccaa aggcccccgt gccactgctc ccacccctgc ccccacctcc acctgagccc
     1381 gagagctccg aggaccccac cagcccccct gagccccagg acttgagcag cagcgtctgc
     1441 aaagaggaga agatgcccag aggaggctca ctggagagcg acggctgccc caaggagcca
     1501 gctaagactc agcccgcggt tgccaccgcc gccacggccg cagaaaagta caaacaccga
     1561 ggggagggag agcgcaaaga cattgtttca tcctccatgc caaggccaaa cagagaggag
     1621 cctgtggaca gccggacgcc cgtgaccgag agagttagct gatagaccca gctttcttgt
     1681 acaaagttgg cattataaaa aataattgct catcaatttg ttgcaacgaa caggtcacta
     1741 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
     1801 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
     1861 ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac agaaatgcct
     1921 cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac
     1981 gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct
     2041 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
     2101 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc
     2161 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
     2221 agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat
     2281 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg
     2341 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt
     2401 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc
     2461 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca
     2521 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg
     2581 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg
     2641 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg
     2701 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct
     2761 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt
     2821 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg
     2881 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg
     2941 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg
     3001 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat
     3061 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa
     3121 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3181 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3241 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3301 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3361 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3421 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3481 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3541 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3601 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3661 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3721 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3781 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3841 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     3901 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     3961 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc
     4021 gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt
     4081 ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg
     4141 ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac
     4201 cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg
     4261 at
//
