LOCUS       sequence-471919-        6819 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express Beta catenin-SmBiT Fusion Protein in Mammalian Cells under a
            CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6819)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6819)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Beta catenin-SmBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6819
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1189..3531
                     /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
                     SRVLKVLSVCSSNKPAIVEAGGMQALGLHLTDPSQRLVQNCLWTLRNLSDAATKQEGME
                     GLLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRTVLRAGDRE
                     DITEPAICALRHLTSRHQEAEMAQNAVRLHYGLPVVVKLLHPPSHWPLIKATVGLIRNL
                     ALCPANHAPLREQGAIPRLVQLLVRAHQDTQRRTSMGGTQQQFVEGVRMEEIVEGCTGA
                     LHILARDVHNRIVIRGLNTIPLFVQLLYSPIENIQRVAAGVLCELAQDKEAAEAIEAEG
                     ATAPLTELLHSRNEGVATYAAAVLFRMSEDKPQDYKKRLSVELTSSLFRTEPMAWNETA
                     DLGLDIGAQGEPLGYRQDDPSYRSFHSGGYGQDALGMDPMMEHEMGGHHPGADYPVDGL
                     PDLGHAQDLMDGLPPGDSNQLAWFDTDL"
     CDS             1189..3531
                     /codon_start=1
                     /gene="Homo sapiens CTNNB1"
                     /product="human cadherin-associated protein that also plays
                     a role in the canonical Wnt signaling pathway"
                     /label=beta-catenin
                     /note="also known as catenin beta-1"
                     /translation="MATQADLMELDMAMEPDRKAAVSHWQQQSYLDSGIHSGATTTAPS
                     LSGKGNPEEEDVDTSQVLYEWEQGFSQSFTQEQVADIDGQYAMTRAQRVRAAMFPETLD
                     EGMQIPSTQFDAAHPTNVQRLAEPSQMLKHAVVNLINYQDDAELATRAIPELTKLLNDE
                     DQVVVNKAAVMVHQLSKKEASRHAIMRSPQMVSAIVRTMQNTNDVETARCTAGTLHNLS
                     HHREGLLAIFKSGGIPALVKMLGSPVDSVLFYAITTLHNLLLHQEGAKMAVRLAGGLQK
                     MVALLNKTNVKFLAITTDCLQILAYGNQESKLIILASGGPQALVNIMRTYTYEKLLWTT
                     SRVLKVLSVCSSNKPAIVEAGGMQALGLHLTDPSQRLVQNCLWTLRNLSDAATKQEGME
                     GLLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRTVLRAGDRE
                     DITEPAICALRHLTSRHQEAEMAQNAVRLHYGLPVVVKLLHPPSHWPLIKATVGLIRNL
                     ALCPANHAPLREQGAIPRLVQLLVRAHQDTQRRTSMGGTQQQFVEGVRMEEIVEGCTGA
                     LHILARDVHNRIVIRGLNTIPLFVQLLYSPIENIQRVAAGVLCELAQDKEAAEAIEAEG
                     ATAPLTELLHSRNEGVATYAAAVLFRMSEDKPQDYKKRLSVELTSSLFRTEPMAWNETA
                     DLGLDIGAQGEPLGYRQDDPSYRSFHSGGYGQDALGMDPMMEHEMGGHHPGADYPVDGL
                     PDLGHAQDLMDGLPPGDSNQLAWFDTDL"
     polyA_signal    3708..3829
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3719..3738)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3944..3977)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3987..4034
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4066..4860
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4120..4139)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4730..4749
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      5077..5622
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     5523..5542
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5739..5756
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    5906..5954
                     /note="synthetic polyadenylation signal"
     CDS             complement(5978..6376)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6373..6382
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6407..6764)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6421..6556
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6573..6764
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6721..6741
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact atagggctag cgatcgccat ggctactcaa
     1201 gctgatttga tggagttgga catggccatg gaaccagaca gaaaagcggc tgttagtcac
     1261 tggcagcaac agtcttacct ggactctgga atccattctg gtgccactac cacagctcct
     1321 tctctgagtg gtaaaggcaa tcctgaggaa gaggatgtgg atacctccca agtcctgtat
     1381 gagtgggaac agggattttc tcagtccttc actcaagaac aagtagctga tattgatgga
     1441 cagtatgcaa tgactcgagc tcagagggta cgagctgcta tgttccctga gacattagat
     1501 gagggcatgc agatcccatc tacacagttt gatgctgctc atcccactaa tgtccagcgt
     1561 ttggctgaac catcacagat gctgaaacat gcagttgtaa acttgattaa ctatcaagat
     1621 gatgcagaac ttgccacacg tgcaatccct gaactgacaa aactgctaaa tgacgaggac
     1681 caggtggtgg ttaataaggc tgcagttatg gtccatcagc tttctaaaaa ggaagcttcc
     1741 agacacgcta tcatgcgttc tcctcagatg gtgtctgcta ttgtacgtac catgcagaat
     1801 acaaatgatg tagaaacagc tcgttgtacc gctgggacct tgcataacct ttcccatcat
     1861 cgtgagggct tactggccat ctttaagtct ggaggcattc ctgccctggt gaaaatgctt
     1921 ggttcaccag tggattctgt gttgttttat gccattacaa ctctccacaa ccttttatta
     1981 catcaagaag gagctaaaat ggcagtgcgt ttagctggtg ggctgcagaa aatggttgcc
     2041 ttgctcaaca aaacaaatgt taaattcttg gctattacga cagactgcct tcaaatttta
     2101 gcttatggca accaagaaag caagctcatc atactggcta gtggtggacc ccaagcttta
     2161 gtaaatataa tgaggaccta tacttacgaa aaactactgt ggaccacaag cagagtgctg
     2221 aaggtgctat ctgtctgctc tagtaataag ccggctattg tagaagctgg tggaatgcaa
     2281 gctttaggac ttcacctgac agatccaagt caacgtcttg ttcagaactg tctttggact
     2341 ctcaggaatc tttcagatgc tgcaactaaa caggaaggga tggaaggtct ccttgggact
     2401 cttgttcagc ttctgggttc agatgatata aatgtggtca cctgtgcagc tggaattctt
     2461 tctaacctca cttgcaataa ttataagaac aagatgatgg tctgccaagt gggtggtata
     2521 gaggctcttg tgcgtactgt ccttcgggct ggtgacaggg aagacatcac tgagcctgcc
     2581 atctgtgctc ttcgtcatct gaccagccga caccaagaag cagagatggc ccagaatgca
     2641 gttcgccttc actatggact accagttgtg gttaagctct tacacccacc atcccactgg
     2701 cctctgataa aggctactgt tggattgatt cgaaatcttg ccctttgtcc cgcaaatcat
     2761 gcacctttgc gtgagcaggg tgccattcca cgactagttc agttgcttgt tcgtgcacat
     2821 caggataccc agcgccgtac gtccatgggt gggacacagc agcaatttgt ggagggggtc
     2881 cgcatggaag aaatagttga aggttgtacc ggagcccttc acatcctagc tcgggatgtt
     2941 cacaaccgaa ttgttatcag aggactaaat accattccat tgtttgtgca gctgctttat
     3001 tctcccattg aaaacatcca aagagtagct gcaggggtcc tctgtgaact tgctcaggac
     3061 aaggaagctg cagaagctat tgaagctgag ggagccacag ctcctctgac agagttactt
     3121 cactctagga atgaaggtgt ggcgacatat gcagctgctg ttttgttccg aatgtctgag
     3181 gacaagccac aagattacaa gaaacggctt tcagttgagc tgaccagctc tctcttcaga
     3241 acagagccaa tggcttggaa tgagactgct gatcttggac ttgatattgg tgcccaggga
     3301 gaaccccttg gatatcgcca ggatgatcct agctatcgtt cttttcactc tggtggatat
     3361 ggccaggatg ccttgggtat ggaccccatg atggaacatg agatgggtgg ccaccaccct
     3421 ggtgctgact atccagttga tgggctgcca gatctggggc atgcccagga cctcatggat
     3481 gggctgcctc caggtgacag caatcagctg gcctggtttg atactgacct ggtttctcaa
     3541 ggcagttcag gtggtggcgg gagcggaggt ggaggctcga gcggtgtgac cggctaccgg
     3601 ctgttcgagg agattctgta aggccgcgac tctagagtcg acctgcaggc atgcaagctg
     3661 atttggctgc tgccaccgct gagcaaggcc gcttcgagca gacatgataa gatacattga
     3721 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3781 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3841 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta
     3901 aaacctctac aaatgtggta aaatcttaat taagaattct aatataactt cgtatagcat
     3961 acattatacg aagttatgga tccactgttg acaattaatc atcggcatag tatatcggca
     4021 tagtataata cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat
     4081 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4141 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4201 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4261 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4321 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4381 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4441 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4501 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4561 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4621 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4681 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4741 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4801 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4861 ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac
     4921 ggttatccac agaatcaggg gataacgcag gaaagaacat gtcaaacatg agaattacaa
     4981 cttatatcgt atggggctga cttcaggtgc tacatttgaa gagataaatt gcactgaaat
     5041 ctagaaatat tttatctgat taataagatg atcttcttga gatcgttttg gtctgcgcgt
     5101 aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga aggttctctg
     5161 agctaccaac tctttgaacc gaggtaactg gcttggagga gcgcagtcac caaaacttgt
     5221 cctttcagtt tagccttaac cggcgcatga cttcaagact aactcctcta aatcaattac
     5281 cagtggctgc tgccagtggt gcttttgcat gtctttccgg gttggactca agacgatagt
     5341 taccggataa ggcgcagcgg tcggactgaa cggggggttc gtgcatacag tccagcttgg
     5401 agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg cggccataac
     5461 agcggaatga caccggtaaa ccgaaaggca ggaacaggag agcgcacgag ggagccgcca
     5521 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg atttgagcgt
     5581 cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt tgccgcggcc
     5641 ctctcacttc cctgttaagt atcttcctgg catcttccag gaaatctccg ccccgttcgt
     5701 aagccatttc cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag
     5761 cggaatatat cctgtatcac atattctgct gacgcaccgg tgcagccttt tttctcctgc
     5821 cacatgaagc acttcactga caccctcatc agtgccaaca tagtaagcca gtatacactc
     5881 cgctagcgct gatgtccgga atcgtcacac aaaaaaccaa cacacagatg taatgaaaat
     5941 aaagatattt tattgcggcc atcgtgatgg ctagagttta gccctcccac acataaccag
     6001 agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc ttcactatgg
     6061 ctttgatccc aggatgcaga tcgagaagca cctgtcggca ccgtccgcag gggctcaaga
     6121 tgcccctgtt ctcatttccg atcgcgacga tacaagtcag gttgccagct gccgcagcag
     6181 cagcagtgcc cagcaccacg agttctgcac aaggtccccc agtaaaatga tatacattga
     6241 caccagtgaa gatgcggccg tcgctagaga gagctgcgct ggcgacgctg tagtcttcag
     6301 agatggggat gctgttgatt gtagccgttg ctctttcaat gagggtggat tcttcttgag
     6361 acaaaggctt ggccatggtg gcgctagtgt cagaagaatc gagctttttg caaaagccta
     6421 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     6481 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     6541 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
     6601 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
     6661 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     6721 accctaactg acacacattc cacagctggt tctttccgcc tcagaaggta cctaaccaag
     6781 ttcctctttc agaggttatt tcaggccatg gtgctgcgc
//
