LOCUS       sequence-480843-        7885 bp    DNA     circular SYN 28-JUL-2025
DEFINITION  Express Beta-Catenin-NanoLuc 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 7885)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7885)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 28, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: Beta-Catenin-NanoLuc Vector
FEATURES             Location/Qualifiers
     source          1..7885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(251..2593)
                     /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             complement(251..2593)
                     /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"
     promoter        complement(2632..2650)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2633..2650)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2770..3073
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3228..3248
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3281..3328
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3360..4019
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3426..3445)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(4191..4779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4271..4290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4905..4953
                     /note="synthetic polyadenylation signal"
     CDS             complement(4977..5771)
                     /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(5088..5107)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5698..5717
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6149..7327)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6158..7096
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(6181..6201)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     polyA_signal    7344..7425
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     7369..7388
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(7602..7885,1..229))
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
ORIGIN
        1 ttttttcgat ctggcccatt tggtcgccgc tcagaccttc atacgggatg atgacatgga
       61 tgtcgatctt cagcccattt tcaccgctca ggacaatcct ttggatcgga gttacggaca
      121 ccccgagatt ctgaaacaaa ctggacacac ctccctgttc aaggacttgg tccaggttgt
      181 agccggctgt ctgtcgccag tccccaacga aatcttcgag tgtgaagacg ccgctcgagc
      241 cgagagaaac caggtcagta tcaaaccagg ccagctgatt gctgtcacct ggaggcagcc
      301 catccatgag gtcctgggca tgccccagat ctggcagccc atcaactgga tagtcagcac
      361 cagggtggtg gccacccatc tcatgttcca tcatggggtc catacccaag gcatcctggc
      421 catatccacc agagtgaaaa gaacgatagc taggatcatc ctggcgatat ccaaggggtt
      481 ctccctgggc accaatatca agtccaagat cagcagtctc attccaagcc attggctctg
      541 ttctgaagag agagctggtc agctcaactg aaagccgttt cttgtaatct tgtggcttgt
      601 cctcagacat tcggaacaaa acagcagctg catatgtcgc cacaccttca ttcctagagt
      661 gaagtaactc tgtcagagga gctgtggctc cctcagcttc aatagcttct gcagcttcct
      721 tgtcctgagc aagttcacag aggacccctg cagctactct ttggatgttt tcaatgggag
      781 aataaagcag ctgcacaaac aatggaatgg tatttagtcc tctgataaca attcggttgt
      841 gaacatcccg agctaggatg tgaagggctc cggtacaacc ttcaactatt tcttccatgc
      901 ggaccccctc cacaaattgc tgctgtgtcc cacccatgga cgtacggcgc tgggtatcct
      961 gatgtgcacg aacaagcaac tgaactagtc gtggaatggc accctgctca cgcaaaggtg
     1021 catgatttgc gggacaaagg gcaagatttc gaatcaatcc aacagtagcc tttatcagag
     1081 gccagtggga tggtgggtgt aagagcttaa ccacaactgg tagtccatag tgaaggcgaa
     1141 ctgcattctg ggccatctct gcttcttggt gtcggctggt cagatgacga agagcacaga
     1201 tggcaggctc agtgatgtct tccctgtcac cagcccgaag gacagtacgc acaagagcct
     1261 ctataccacc cacttggcag accatcatct tgttcttata attattgcaa gtgaggttag
     1321 aaagaattcc agctgcacag gtgaccacat ttatatcatc tgaacccaga agctgaacaa
     1381 gagtcccaag gagaccttcc atcccttcct gtttagttgc agcatctgaa agattcctga
     1441 gagtccaaag acagttctga acaagacgtt gacttggatc tgtcaggtga agtcctaaag
     1501 cttgcattcc accagcttct acaatagccg gcttattact agagcagaca gatagcacct
     1561 tcagcactct gcttgtggtc cacagtagtt tttcgtaagt ataggtcctc attatattta
     1621 ctaaagcttg gggtccacca ctagccagta tgatgagctt gctttcttgg ttgccataag
     1681 ctaaaatttg aaggcagtct gtcgtaatag ccaagaattt aacatttgtt ttgttgagca
     1741 aggcaaccat tttctgcagc ccaccagcta aacgcactgc cattttagct ccttcttgat
     1801 gtaataaaag gttgtggaga gttgtaatgg cataaaacaa cacagaatcc actggtgaac
     1861 caagcatttt caccagggca ggaatgcctc cagacttaaa gatggccagt aagccctcac
     1921 gatgatggga aaggttatgc aaggtcccag cggtacaacg agctgtttct acatcatttg
     1981 tattctgcat ggtacgtaca atagcagaca ccatctgagg agaacgcatg atagcgtgtc
     2041 tggaagcttc ctttttagaa agctgatgga ccataactgc agccttatta accaccacct
     2101 ggtcctcgtc atttagcagt tttgtcagtt cagggattgc acgtgtggca agttctgcat
     2161 catcttgata gttaatcaag tttacaactg catgtttcag catctgtgat ggttcagcca
     2221 aacgctggac attagtggga tgagcagcat caaactgtgt agatgggatc tgcatgccct
     2281 catctaatgt ctcagggaac atagcagctc gtaccctctg agctcgagtc attgcatact
     2341 gtccatcaat atcagctact tgttcttgag tgaaggactg agaaaatccc tgttcccact
     2401 catacaggac ttgggaggta tccacatcct cttcctcagg attgccttta ccactcagag
     2461 aaggagctgt ggtagtggca ccagaatgga ttccagagtc caggtaagac tgttgctgcc
     2521 agtgactaac agccgctttt ctgtctggtt ccatggccat gtccaactcc atcaaatcag
     2581 cttgagtagc catggcgatc gctagctgca aaaagaacaa gtagcttgta ttctatagtg
     2641 tcacctaaat gggcgatctg acggttcact aaaccagctc tgcttatata ggcctccctg
     2701 gcgcgccact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     2761 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     2821 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     2881 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     2941 tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     3001 tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat
     3061 cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga
     3121 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     3181 aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg
     3241 taggcgtgta cggtgggagg tcaaatcgaa ttctaatact gttgacaatt aatcatcggc
     3301 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca
     3361 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
     3421 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
     3481 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
     3541 acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg
     3601 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa
     3661 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt
     3721 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga
     3781 atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg
     3841 ccaatatgga caacttcttc gcccccgttt tcactatggg caaatattat acgcaaggcg
     3901 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtctgtgat ggcttccatg
     3961 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaag
     4021 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aaccggtatc agctcactca
     4081 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4141 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4201 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4261 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4321 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4381 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4441 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4501 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4561 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4621 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4681 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4741 tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt gatcttttct
     4801 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4861 tcaaaaagga tcttcaccta gatcctttta tagtccggaa tcgtcacaca aaaaaccaac
     4921 acacagatgt aatgaaaata aagatatttt attgcggcca tcgtgatggc tagagttcag
     4981 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg
     5041 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta
     5101 gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca
     5161 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccgtg ggtcacgacg
     5221 agatcctcgc cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc
     5281 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt
     5341 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta
     5401 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat
     5461 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg
     5521 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct
     5581 gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
     5641 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
     5701 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
     5761 tgttcaatca tggtggcatc gattcccggt caacatccag tacacctccg gcacaccggc
     5821 ttccccaagg gcgccacgct ctcccaccac aacatcctca acaacggcta ctgggtgggc
     5881 cgcacggtcg gctacacgga gcaggaccgg gtgtgtctgc cggtgccctt ctaccactgc
     5941 ttcggcatgg tcatgggcaa cttgggcgcc acctcccacg ccgcctgcat cgtcatcccg
     6001 gccccgtcct tcgagccggc ggcgacgctg gaggcggtgc agcgggagag gtgcacgtcc
     6061 ctgtacggcg tccccacgaa gccgaattcc accacactgg actagatgca tgctcgacag
     6121 tgtgatggat atctgcagaa ttcggctttc acgacacctg aaatggaaga aaaaaacttt
     6181 gaaccactgt ctgaggcttg agaatgaacc aagatccaaa ctcaaaaagg gcaaattcca
     6241 aggagaatta catcaagtgc caagctggcc taacttcagt ctccacccac tcagtgtggg
     6301 gaaactccat cgcataaaac ccctcccccc aacctaaaga cgacgtactc caaaagctcg
     6361 agaactaatc gaggtgcctg gacggcgccc ggtactccgt ggagtcacat gaagcgacgg
     6421 ctgaggacgg aaaggccctt ttcctttgtg tgggtgactc acccgcccgc tctcccgagc
     6481 gccgcgtcct ccattttgag ctgcctgcag cagggccggg aagcggccat ctttccgctc
     6541 acgcaactgg tgccgaccgg gccagccttg ccgcccaggg cggggcgata cacggcggcg
     6601 cgaggccagg caccagagca ggccggccag cttgagacta cccccgtccg attctcggtg
     6661 gccgcgctcg caggccccgc ctcgccgaac atgtgcgctg ggacgcacgg gccccgtcgc
     6721 cgcccgcggc cccaaaaacc gaaataccag tgtgcagatc ttggcccgca tttacaagac
     6781 tatcttgcca gaaaaaaagc gtcgcagcag gtcatcaaaa attttaaatg gctagagact
     6841 tatcgaaagc agcgagacag gcgcgaaggt gccaccagat tcgcacgcgg cggccccagc
     6901 gcccaggcca ggcctcaact caagcacgag gcgaaggggc tccttaagcg caaggcctcg
     6961 aactctccca cccacttcca acccgaagct cgggatcaag aatcacgtac tgcagccagg
     7021 tggaagtaat tcaaggcacg caagggccat aacccgtaaa gaggccaggc ccgcgggaac
     7081 cacacacggc acttacctgt gttctggcgg caaacccgtt gcgaaaaaga acgttcacgg
     7141 cgactactgc acttatatac ggttctcccc caccctcggg aaaaaggcgg agccagtaca
     7201 cgacatcact ttcccagttt accccgcgcc accttctcta ggcaccggtt caattgccga
     7261 cccctccccc caacttctcg gggactgtgg gcgatgtgcg ctctgcccac tgacgggcac
     7321 cggagccaag acgcgtgacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
     7381 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     7441 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     7501 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
     7561 tgacttcgta cgagagccta ggtgtttcta gagtcgcggc cttacgccag aatgcgttcg
     7621 cacagccgcc agccggtcac tccgttgatg gttactcgga acagcaggga gccgtcgggg
     7681 ttgatcaggc gctcgtcgat aattttgttg ccgttccaca gggtccctgt tacagtgatc
     7741 tttttgccgt cgaacacggc gatgccttca tacggccgtc cgaaatagtc gatcatgttc
     7801 ggcgtaaccc cgtcgattac cagtgtgcca tagtgcagga tcaccttaaa gtgatgatca
     7861 tccacagggt acaccacctt aaaaa
//
