LOCUS       sequence-427250-        5147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway cloning donor vector containing CTNNB1 ASD mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5147)
  AUTHORS   Allan, Loewen Labs
  TITLE     Allan and Loewen Labs SFARI funded autism variants
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5147)
  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            "Unpublished"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: CTNNB1 - R376H in pENTR
FEATURES             Location/Qualifiers
     source          1..5147
                     /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             202..2547
                     /codon_start=1
                     /gene="CTNNB1"
                     /product="protein involved both in cell-cell adhesion as 
                     part of the cadherin protein complex, and in gene 
                     transcription as part of the Wnt signaling pathway"
                     /label=CTNNB1
                     /note="also known as catenin beta-1; related to Drosophila 
                     armadillo"
                     /translation="MATQADLMELDMAMEPDRKAAVSHWQQQSYLDSGIHSGATTTAPS
                     LSGKGNPEEEDVDTSQVLYEWEQGFSQSFTQEQVADIDGQYAMTRAQRVRAAMFPETLD
                     EGMQIPSTQFDAAHPTNVQRLAEPSQMLKHAVVNLINYQDDAELATRAIPELTKLLNDE
                     DQVVVNKAAVMVHQLSKKEASRHAIMRSPQMVSAIVRTMQNTNDVETARCTAGTLHNLS
                     HHREGLLAIFKSGGIPALVKMLGSPVDSVLFYAITTLHNLLLHQEGAKMAVRLAGGLQK
                     MVALLNKTNVKFLAITTDCLQILAYGNQESKLIILASGGPQALVNIMRTYTYEKLLWTT
                     SRVLKVLSVCSSNKPAIVEAGGMQALGLHLTDPSQHLVQNCLWTLRNLSDAATKQEGME
                     GLLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRTVLRAGDRE
                     DITEPAICALRHLTSRHQEAEMAQNAVRLHYGLPVVVKLLHPPSHWPLIKATVGLIRNL
                     ALCPANHAPLREQGAIPRLVQLLVRAHQDTQRRTSMGGTQQQFVEGVRMEEIVEGCTGA
                     LHILARDVHNRIVIRGLNTIPLFVQLLYSPIENIQRVAAGVLCELAQDKEAAEAIEAEG
                     ATAPLTELLHSRNEGVATYAAAVLFRMSEDKPQDYKKRLSVELTSSLFRTEPMAWNETA
                     DLGLDIGAQGEPLGYRQDDPSYRSFHSGGYGQDALGMDPMMEHEMGGHHPGADYPVDGL
                     PDLGHAQDLMDGLPPGDSNQLAWFDTDL"
     protein_bind    complement(2551..2650)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2667..2686)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2668..2686)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2691..2707)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2691..2707)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3138..3929
                     /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      4022..4610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4511..4530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4764..4781
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4940..4967)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5059..5145)
                     /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 catggctact caagctgatt tgatggagtt ggacatggcc
      241 atggaaccag acagaaaagc ggctgttagt cactggcagc aacagtctta cctggactct
      301 ggaatccatt ctggtgccac taccacagct ccttctctga gtggtaaagg caatcctgag
      361 gaagaggatg tggatacctc ccaagtcctg tatgagtggg aacagggatt ttctcagtcc
      421 ttcactcaag aacaagtagc tgatattgat ggacagtatg caatgactcg agctcagagg
      481 gtacgagctg ctatgttccc tgagacatta gatgagggca tgcagatccc atctacacag
      541 tttgatgctg ctcatcccac taatgtccag cgtttggctg aaccatcaca gatgctgaaa
      601 catgcagttg taaacttgat taactatcaa gatgatgcag aacttgccac acgtgcaatc
      661 cctgaactga caaaactgct aaatgacgag gaccaggtgg tggttaataa ggctgcagtt
      721 atggtccatc agctttctaa aaaggaagct tccagacacg ctatcatgcg ttctcctcag
      781 atggtgtctg ctattgtacg taccatgcag aatacaaatg atgtagaaac agctcgttgt
      841 accgctggga ccttgcataa cctttcccat catcgtgagg gcttactggc catctttaag
      901 tctggaggca ttcctgccct ggtgaaaatg cttggttcac cagtggattc tgtgttgttt
      961 tatgccatta caactctcca caacctttta ttacatcaag aaggagctaa aatggcagtg
     1021 cgtttagctg gtgggctgca gaaaatggtt gccttgctca acaaaacaaa tgttaaattc
     1081 ttggctatta cgacagactg ccttcaaatt ttagcttatg gcaaccaaga aagcaagctc
     1141 atcatactgg ctagtggtgg accccaagct ttagtaaata taatgaggac ctatacttac
     1201 gaaaaactac tgtggaccac aagcagagtg ctgaaggtgc tatctgtctg ctctagtaat
     1261 aagccggcta ttgtagaagc tggtggaatg caagctttag gacttcacct gacagatcca
     1321 agtcaacatc ttgttcagaa ctgtctttgg actctcagga atctttcaga tgctgcaact
     1381 aaacaggaag ggatggaagg tctccttggg actcttgttc agcttctggg ttcagatgat
     1441 ataaatgtgg tcacctgtgc agctggaatt ctttctaacc tcacttgcaa taattataag
     1501 aacaagatga tggtctgcca agtgggtggt atagaggctc ttgtgcgtac tgtccttcgg
     1561 gctggtgaca gggaagacat cactgagcct gccatctgtg ctcttcgtca tctgaccagc
     1621 cgacaccaag aagcagagat ggcccagaat gcagttcgcc ttcactatgg actaccagtt
     1681 gtggttaagc tcttacaccc accatcccac tggcctctga taaaggctac tgttggattg
     1741 attcgaaatc ttgccctttg tcccgcaaat catgcacctt tgcgtgagca gggtgccatt
     1801 ccacgactag ttcagttgct tgttcgtgca catcaggata cccagcgccg tacgtccatg
     1861 ggtgggacac agcagcaatt tgtggagggg gtccgcatgg aagaaatagt tgaaggttgt
     1921 accggagccc ttcacatcct agctcgggat gttcacaacc gaattgttat cagaggacta
     1981 aataccattc cattgtttgt gcagctgctt tattctccca ttgaaaacat ccaaagagta
     2041 gctgcagggg tcctctgtga acttgctcag gacaaggaag ctgcagaagc tattgaagct
     2101 gagggagcca cagctcctct gacagagtta cttcactcta ggaatgaagg tgtggcgaca
     2161 tatgcagctg ctgttttgtt ccgaatgtct gaggacaagc cacaagatta caagaaacgg
     2221 ctttcagttg agctgaccag ctctctcttc agaacagagc caatggcttg gaatgagact
     2281 gctgatcttg gacttgatat tggtgcccag ggagaacccc ttggatatcg ccaggatgat
     2341 cctagctatc gttcttttca ctctggtgga tatggccagg atgccttggg tatggacccc
     2401 atgatggaac atgagatggg tggccaccac cctggtgctg actatccagt tgatgggctg
     2461 ccagatctgg ggcatgccca ggacctcatg gatgggctgc ctccaggtga cagcaatcag
     2521 ctggcctggt ttgatactga cctgtaatag acccagcttt cttgtacaaa gttggcatta
     2581 taaaaaataa ttgctcatca atttgttgca acgaacaggt cactatcagt caaaataaaa
     2641 tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg
     2701 tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     2761 atatatcatc atgcctcctc tagaccagcc aggacagaaa tgcctcgact tcgctgctac
     2821 ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat
     2881 aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca
     2941 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
     3001 actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg
     3061 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta
     3121 aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag
     3181 aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg
     3241 gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga
     3301 ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg
     3361 cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg
     3421 acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca
     3481 atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc
     3541 tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg
     3601 atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact
     3661 cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt
     3721 acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc
     3781 gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag
     3841 aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag
     3901 gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta
     3961 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4021 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4081 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4141 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4201 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4261 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4321 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4381 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4441 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4501 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4561 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4621 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4681 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4741 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     4801 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     4861 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg
     4921 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg
     4981 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt
     5041 caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
     5101 aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgat
//
