LOCUS       sequence-471918-        6878 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express SmBiT-Beta catenin 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 6878)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6878)
  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: SmBiT-Beta catenin Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6878
                     /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             1279..3627
                     /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
                     PDLGHAQDLMDGLPPGDSNQLAWFDTDLV"
     polyA_signal    3767..3888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3778..3797)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4003..4036)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4046..4093
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4125..4919
                     /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(4179..4198)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4789..4808
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      5136..5681
                     /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     5582..5601
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5798..5815
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    5965..6013
                     /note="synthetic polyadenylation signal"
     CDS             complement(6037..6435)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6432..6441
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6466..6823)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6480..6615
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6632..6823
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6780..6800
                     /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 cgctcaccat ggtgaccggc
     1201 taccggctgt tcgaggagat tctcgggagc tccggtggtg gcgggagcgg aggtggaggc
     1261 tcgagcggtg cgatcgccat ggctactcaa gctgatttga tggagttgga catggccatg
     1321 gaaccagaca gaaaagcggc tgttagtcac tggcagcaac agtcttacct ggactctgga
     1381 atccattctg gtgccactac cacagctcct tctctgagtg gtaaaggcaa tcctgaggaa
     1441 gaggatgtgg atacctccca agtcctgtat gagtgggaac agggattttc tcagtccttc
     1501 actcaagaac aagtagctga tattgatgga cagtatgcaa tgactcgagc tcagagggta
     1561 cgagctgcta tgttccctga gacattagat gagggcatgc agatcccatc tacacagttt
     1621 gatgctgctc atcccactaa tgtccagcgt ttggctgaac catcacagat gctgaaacat
     1681 gcagttgtaa acttgattaa ctatcaagat gatgcagaac ttgccacacg tgcaatccct
     1741 gaactgacaa aactgctaaa tgacgaggac caggtggtgg ttaataaggc tgcagttatg
     1801 gtccatcagc tttctaaaaa ggaagcttcc agacacgcta tcatgcgttc tcctcagatg
     1861 gtgtctgcta ttgtacgtac catgcagaat acaaatgatg tagaaacagc tcgttgtacc
     1921 gctgggacct tgcataacct ttcccatcat cgtgagggct tactggccat ctttaagtct
     1981 ggaggcattc ctgccctggt gaaaatgctt ggttcaccag tggattctgt gttgttttat
     2041 gccattacaa ctctccacaa ccttttatta catcaagaag gagctaaaat ggcagtgcgt
     2101 ttagctggtg ggctgcagaa aatggttgcc ttgctcaaca aaacaaatgt taaattcttg
     2161 gctattacga cagactgcct tcaaatttta gcttatggca accaagaaag caagctcatc
     2221 atactggcta gtggtggacc ccaagcttta gtaaatataa tgaggaccta tacttacgaa
     2281 aaactactgt ggaccacaag cagagtgctg aaggtgctat ctgtctgctc tagtaataag
     2341 ccggctattg tagaagctgg tggaatgcaa gctttaggac ttcacctgac agatccaagt
     2401 caacgtcttg ttcagaactg tctttggact ctcaggaatc tttcagatgc tgcaactaaa
     2461 caggaaggga tggaaggtct ccttgggact cttgttcagc ttctgggttc agatgatata
     2521 aatgtggtca cctgtgcagc tggaattctt tctaacctca cttgcaataa ttataagaac
     2581 aagatgatgg tctgccaagt gggtggtata gaggctcttg tgcgtactgt ccttcgggct
     2641 ggtgacaggg aagacatcac tgagcctgcc atctgtgctc ttcgtcatct gaccagccga
     2701 caccaagaag cagagatggc ccagaatgca gttcgccttc actatggact accagttgtg
     2761 gttaagctct tacacccacc atcccactgg cctctgataa aggctactgt tggattgatt
     2821 cgaaatcttg ccctttgtcc cgcaaatcat gcacctttgc gtgagcaggg tgccattcca
     2881 cgactagttc agttgcttgt tcgtgcacat caggataccc agcgccgtac gtccatgggt
     2941 gggacacagc agcaatttgt ggagggggtc cgcatggaag aaatagttga aggttgtacc
     3001 ggagcccttc acatcctagc tcgggatgtt cacaaccgaa ttgttatcag aggactaaat
     3061 accattccat tgtttgtgca gctgctttat tctcccattg aaaacatcca aagagtagct
     3121 gcaggggtcc tctgtgaact tgctcaggac aaggaagctg cagaagctat tgaagctgag
     3181 ggagccacag ctcctctgac agagttactt cactctagga atgaaggtgt ggcgacatat
     3241 gcagctgctg ttttgttccg aatgtctgag gacaagccac aagattacaa gaaacggctt
     3301 tcagttgagc tgaccagctc tctcttcaga acagagccaa tggcttggaa tgagactgct
     3361 gatcttggac ttgatattgg tgcccaggga gaaccccttg gatatcgcca ggatgatcct
     3421 agctatcgtt cttttcactc tggtggatat ggccaggatg ccttgggtat ggaccccatg
     3481 atggaacatg agatgggtgg ccaccaccct ggtgctgact atccagttga tgggctgcca
     3541 gatctggggc atgcccagga cctcatggat gggctgcctc caggtgacag caatcagctg
     3601 gcctggtttg atactgacct ggtttaaacg aattcgggct cggtacccgg ggatcctcta
     3661 gagtcgacct gcaggcatgc aagctgatcc ggctgctaac aaagcccgaa aggaagctga
     3721 gttggctgct gccaccgctg agcaaggccg cttcgagcag acatgataag atacattgat
     3781 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     3841 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     3901 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt aagcaagtaa
     3961 aacctctaca aatgtggtaa aatcttaatt aagaattcta atataacttc gtatagcata
     4021 cattatacga agttatggat ccactgttga caattaatca tcggcatagt atatcggcat
     4081 agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgatt gaacaagatg
     4141 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4201 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4261 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4321 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4381 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4441 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4501 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4561 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4621 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4681 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4741 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4801 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4861 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgat
     4921 tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg cggtaatacg
     4981 gttatccaca gaatcagggg ataacgcagg aaagaacatg tcaaacatga gaattacaac
     5041 ttatatcgta tggggctgac ttcaggtgct acatttgaag agataaattg cactgaaatc
     5101 tagaaatatt ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta
     5161 atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga
     5221 gctaccaact ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc
     5281 ctttcagttt agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc
     5341 agtggctgct gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt
     5401 accggataag gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga
     5461 gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca
     5521 gcggaatgac accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag
     5581 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc
     5641 agatttcgtg atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc
     5701 tctcacttcc ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta
     5761 agccatttcc gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc
     5821 ggaatatatc ctgtatcaca tattctgctg acgcaccggt gcagcctttt ttctcctgcc
     5881 acatgaagca cttcactgac accctcatca gtgccaacat agtaagccag tatacactcc
     5941 gctagcgctg atgtccggaa tcgtcacaca aaaaaccaac acacagatgt aatgaaaata
     6001 aagatatttt attgcggcca tcgtgatggc tagagtttag ccctcccaca cataaccaga
     6061 gggcagcaat tcacgaatcc caactgccgt cggctgtcca tcactgtcct tcactatggc
     6121 tttgatccca ggatgcagat cgagaagcac ctgtcggcac cgtccgcagg ggctcaagat
     6181 gcccctgttc tcatttccga tcgcgacgat acaagtcagg ttgccagctg ccgcagcagc
     6241 agcagtgccc agcaccacga gttctgcaca aggtccccca gtaaaatgat atacattgac
     6301 accagtgaag atgcggccgt cgctagagag agctgcgctg gcgacgctgt agtcttcaga
     6361 gatggggatg ctgttgattg tagccgttgc tctttcaatg agggtggatt cttcttgaga
     6421 caaaggcttg gccatggtgg cgctagtgtc agaagaatcg agctttttgc aaaagcctag
     6481 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc
     6541 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     6601 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     6661 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     6721 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     6781 ccctaactga cacacattcc acagctggtt ctttccgcct cagaaggtac ctaaccaagt
     6841 tcctctttca gaggttattt caggccatgg tgctgcgc
//
