LOCUS       sequence-481949-        7911 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Express NanoLuc-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 7911)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7911)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-beta-Catenin Vector
FEATURES             Location/Qualifiers
     source          1..7911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(295..2643)
                     /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"
     CDS             complement(2665..3177)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     promoter        complement(3216..3234)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3217..3234)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        3354..3657
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3812..3832
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3865..3912
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3944..4603
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4010..4029)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(4775..5363)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4855..4874)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5489..5537
                     /note="synthetic polyadenylation signal"
     CDS             complement(5561..6355)
                     /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(5672..5691)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6282..6301
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6733..7911)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6742..7680
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(6765..6785)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagaggatc cccgggtacc gagcccgaat tcgtttaaac
      301 caggtcagta tcaaaccagg ccagctgatt gctgtcacct ggaggcagcc catccatgag
      361 gtcctgggca tgccccagat ctggcagccc atcaactgga tagtcagcac cagggtggtg
      421 gccacccatc tcatgttcca tcatggggtc catacccaag gcatcctggc catatccacc
      481 agagtgaaaa gaacgatagc taggatcatc ctggcgatat ccaaggggtt ctccctgggc
      541 accaatatca agtccaagat cagcagtctc attccaagcc attggctctg ttctgaagag
      601 agagctggtc agctcaactg aaagccgttt cttgtaatct tgtggcttgt cctcagacat
      661 tcggaacaaa acagcagctg catatgtcgc cacaccttca ttcctagagt gaagtaactc
      721 tgtcagagga gctgtggctc cctcagcttc aatagcttct gcagcttcct tgtcctgagc
      781 aagttcacag aggacccctg cagctactct ttggatgttt tcaatgggag aataaagcag
      841 ctgcacaaac aatggaatgg tatttagtcc tctgataaca attcggttgt gaacatcccg
      901 agctaggatg tgaagggctc cggtacaacc ttcaactatt tcttccatgc ggaccccctc
      961 cacaaattgc tgctgtgtcc cacccatgga cgtacggcgc tgggtatcct gatgtgcacg
     1021 aacaagcaac tgaactagtc gtggaatggc accctgctca cgcaaaggtg catgatttgc
     1081 gggacaaagg gcaagatttc gaatcaatcc aacagtagcc tttatcagag gccagtggga
     1141 tggtgggtgt aagagcttaa ccacaactgg tagtccatag tgaaggcgaa ctgcattctg
     1201 ggccatctct gcttcttggt gtcggctggt cagatgacga agagcacaga tggcaggctc
     1261 agtgatgtct tccctgtcac cagcccgaag gacagtacgc acaagagcct ctataccacc
     1321 cacttggcag accatcatct tgttcttata attattgcaa gtgaggttag aaagaattcc
     1381 agctgcacag gtgaccacat ttatatcatc tgaacccaga agctgaacaa gagtcccaag
     1441 gagaccttcc atcccttcct gtttagttgc agcatctgaa agattcctga gagtccaaag
     1501 acagttctga acaagacgtt gacttggatc tgtcaggtga agtcctaaag cttgcattcc
     1561 accagcttct acaatagccg gcttattact agagcagaca gatagcacct tcagcactct
     1621 gcttgtggtc cacagtagtt tttcgtaagt ataggtcctc attatattta ctaaagcttg
     1681 gggtccacca ctagccagta tgatgagctt gctttcttgg ttgccataag ctaaaatttg
     1741 aaggcagtct gtcgtaatag ccaagaattt aacatttgtt ttgttgagca aggcaaccat
     1801 tttctgcagc ccaccagcta aacgcactgc cattttagct ccttcttgat gtaataaaag
     1861 gttgtggaga gttgtaatgg cataaaacaa cacagaatcc actggtgaac caagcatttt
     1921 caccagggca ggaatgcctc cagacttaaa gatggccagt aagccctcac gatgatggga
     1981 aaggttatgc aaggtcccag cggtacaacg agctgtttct acatcatttg tattctgcat
     2041 ggtacgtaca atagcagaca ccatctgagg agaacgcatg atagcgtgtc tggaagcttc
     2101 ctttttagaa agctgatgga ccataactgc agccttatta accaccacct ggtcctcgtc
     2161 atttagcagt tttgtcagtt cagggattgc acgtgtggca agttctgcat catcttgata
     2221 gttaatcaag tttacaactg catgtttcag catctgtgat ggttcagcca aacgctggac
     2281 attagtggga tgagcagcat caaactgtgt agatgggatc tgcatgccct catctaatgt
     2341 ctcagggaac atagcagctc gtaccctctg agctcgagtc attgcatact gtccatcaat
     2401 atcagctact tgttcttgag tgaaggactg agaaaatccc tgttcccact catacaggac
     2461 ttgggaggta tccacatcct cttcctcagg attgccttta ccactcagag aaggagctgt
     2521 ggtagtggca ccagaatgga ttccagagtc caggtaagac tgttgctgcc agtgactaac
     2581 agccgctttt ctgtctggtt ccatggccat gtccaactcc atcaaatcag cttgagtagc
     2641 catggcgatc gcgccgctcg agcccgccag aatgcgttcg cacagccgcc agccggtcac
     2701 tccgttgatg gttactcgga acagcaggga gccgtcgggg ttgatcaggc gctcgtcgat
     2761 aattttgttg ccgttccaca gggtccctgt tacagtgatc tttttgccgt cgaacacggc
     2821 gatgccttca tacggccgtc cgaaatagtc gatcatgttc ggcgtaaccc cgtcgattac
     2881 cagtgtgcca tagtgcagga tcaccttaaa gtgatgatca tccacagggt acaccacctt
     2941 aaaaattttt tcgatctggc ccatttggtc gccgctcaga ccttcatacg ggatgatgac
     3001 atggatgtcg atcttcagcc cattttcacc gctcaggaca atcctttgga tcggagttac
     3061 ggacaccccg agattctgaa acaaactgga cacacctccc tgttcaagga cttggtccag
     3121 gttgtagccg gctgtctgtc gccagtcccc aacgaaatct tcgagtgtga agaccatggt
     3181 gagcgctagc tgcaaaaaga acaagtagct tgtattctat agtgtcacct aaatgggcga
     3241 tctgacggtt cactaaacca gctctgctta tataggcctc cctggcgcgc cactagttat
     3301 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     3361 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     3421 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     3481 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtccg
     3541 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     3601 ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     3661 atgcggtttt ggcagtacac caatgggcgt ggatagcggt ttgactcacg gggatttcca
     3721 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     3781 ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg
     3841 gaggtcaaat cgaattctaa tactgttgac aattaatcat cggcatagta tatcggcata
     3901 gtataatacg acaaggtgag gaactaaacc caggaggcag atcatggaga aaaaaatcac
     3961 tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca
     4021 gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa
     4081 gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct
     4141 gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg tgatatggga
     4201 tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg
     4261 gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg
     4321 ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc
     4381 agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt
     4441 cttcgccccc gttttcacta tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc
     4501 gctggcgatt caggttcatc atgccgtctg tgatggcttc catgtcggca gaatgcttaa
     4561 tgaattacaa cagtactgcg atgagtggca gggcggggcg taagcgggac tctggggttc
     4621 gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt
     4681 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4741 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4801 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4861 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4921 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4981 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5041 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5101 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5161 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     5221 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5281 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5341 cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5401 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5461 cctagatcct tttatagtcc ggaatcgtca cacaaaaaac caacacacag atgtaatgaa
     5521 aataaagata ttttattgcg gccatcgtga tggctagagt tcagaagaac tcgtcaagaa
     5581 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
     5641 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
     5701 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
     5761 ccaccatgat attcggcaag caggcatcgc cgtgggtcac gacgagatcc tcgccgtcgg
     5821 gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
     5881 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
     5941 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
     6001 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
     6061 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
     6121 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt
     6181 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
     6241 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
     6301 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatggtgg
     6361 catcgattcc cggtcaacat ccagtacacc tccggcacac cggcttcccc aagggcgcca
     6421 cgctctccca ccacaacatc ctcaacaacg gctactgggt gggccgcacg gtcggctaca
     6481 cggagcagga ccgggtgtgt ctgccggtgc ccttctacca ctgcttcggc atggtcatgg
     6541 gcaacttggg cgccacctcc cacgccgcct gcatcgtcat cccggccccg tccttcgagc
     6601 cggcggcgac gctggaggcg gtgcagcggg agaggtgcac gtccctgtac ggcgtcccca
     6661 cgaagccgaa ttccaccaca ctggactaga tgcatgctcg acagtgtgat ggatatctgc
     6721 agaattcggc tttcacgaca cctgaaatgg aagaaaaaaa ctttgaacca ctgtctgagg
     6781 cttgagaatg aaccaagatc caaactcaaa aagggcaaat tccaaggaga attacatcaa
     6841 gtgccaagct ggcctaactt cagtctccac ccactcagtg tggggaaact ccatcgcata
     6901 aaacccctcc ccccaaccta aagacgacgt actccaaaag ctcgagaact aatcgaggtg
     6961 cctggacggc gcccggtact ccgtggagtc acatgaagcg acggctgagg acggaaaggc
     7021 ccttttcctt tgtgtgggtg actcacccgc ccgctctccc gagcgccgcg tcctccattt
     7081 tgagctgcct gcagcagggc cgggaagcgg ccatctttcc gctcacgcaa ctggtgccga
     7141 ccgggccagc cttgccgccc agggcggggc gatacacggc ggcgcgaggc caggcaccag
     7201 agcaggccgg ccagcttgag actacccccg tccgattctc ggtggccgcg ctcgcaggcc
     7261 ccgcctcgcc gaacatgtgc gctgggacgc acgggccccg tcgccgcccg cggccccaaa
     7321 aaccgaaata ccagtgtgca gatcttggcc cgcatttaca agactatctt gccagaaaaa
     7381 aagcgtcgca gcaggtcatc aaaaatttta aatggctaga gacttatcga aagcagcgag
     7441 acaggcgcga aggtgccacc agattcgcac gcggcggccc cagcgcccag gccaggcctc
     7501 aactcaagca cgaggcgaag gggctcctta agcgcaaggc ctcgaactct cccacccact
     7561 tccaacccga agctcgggat caagaatcac gtactgcagc caggtggaag taattcaagg
     7621 cacgcaaggg ccataacccg taaagaggcc aggcccgcgg gaaccacaca cggcacttac
     7681 ctgtgttctg gcggcaaacc cgttgcgaaa aagaacgttc acggcgacta ctgcacttat
     7741 atacggttct cccccaccct cgggaaaaag gcggagccag tacacgacat cactttccca
     7801 gtttaccccg cgccaccttc tctaggcacc ggttcaattg ccgacccctc cccccaactt
     7861 ctcggggact gtgggcgatg tgcgctctgc ccactgacgg gcaccggagc c
//
