LOCUS       sequence-471917-        7307 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express Beta catenin-LgBiT 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 7307)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 7307)
  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-LgBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7307
                     /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(806..3148)
                     /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(806..3148)
                     /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(3187..3205)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3188..3205)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        3325..3628
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3783..3803
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3836..3883
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3915..4709
                     /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(3969..3988)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4579..4598
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      4926..5471
                     /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     5372..5391
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5588..5605
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    5755..5803
                     /note="synthetic polyadenylation signal"
     CDS             complement(5827..6864)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5840..5848
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(6895..7252)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6909..7044
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        7061..7252
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7209..7229
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
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 ctagagtcgc ggccttagct gttgatggtt actcggaaca
      301 gcatggagcc gtcgggggtg atcaggcgct cgtcgataat tttgttgccg ttccacaggg
      361 tccctgttac agtgatcttt ttgccgtcga acacggcgat gccttcatac ggccgtccga
      421 aatagttcag catgttcggc gtaaccccgt cgattaccag tgtgccatag ggcaggatca
      481 ccttaaagtg atgatcatcc acagggtaca ccaccttaaa cacctcttcg atctgggcca
      541 tttggtcggc gctcagacct tcatacggga tgatgacatg gatgtcgatc ttcagggcat
      601 tttcaccgct ccggacaatc ctttggatcg gagttacgga cacggcgaga ttctgcagca
      661 aactggacac acctccctgt tcaaggactt ggtccaggtt gtaggcggct gtctgttccc
      721 agtccccaac gaaatcttcg agtgtgaaga caccgctcga gcctccacct ccgctcccgc
      781 caccacctga actgccttga gaaaccaggt cagtatcaaa ccaggccagc tgattgctgt
      841 cacctggagg cagcccatcc atgaggtcct gggcatgccc cagatctggc agcccatcaa
      901 ctggatagtc agcaccaggg tggtggccac ccatctcatg ttccatcatg gggtccatac
      961 ccaaggcatc ctggccatat ccaccagagt gaaaagaacg atagctagga tcatcctggc
     1021 gatatccaag gggttctccc tgggcaccaa tatcaagtcc aagatcagca gtctcattcc
     1081 aagccattgg ctctgttctg aagagagagc tggtcagctc aactgaaagc cgtttcttgt
     1141 aatcttgtgg cttgtcctca gacattcgga acaaaacagc agctgcatat gtcgccacac
     1201 cttcattcct agagtgaagt aactctgtca gaggagctgt ggctccctca gcttcaatag
     1261 cttctgcagc ttccttgtcc tgagcaagtt cacagaggac ccctgcagct actctttgga
     1321 tgttttcaat gggagaataa agcagctgca caaacaatgg aatggtattt agtcctctga
     1381 taacaattcg gttgtgaaca tcccgagcta ggatgtgaag ggctccggta caaccttcaa
     1441 ctatttcttc catgcggacc ccctccacaa attgctgctg tgtcccaccc atggacgtac
     1501 ggcgctgggt atcctgatgt gcacgaacaa gcaactgaac tagtcgtgga atggcaccct
     1561 gctcacgcaa aggtgcatga tttgcgggac aaagggcaag atttcgaatc aatccaacag
     1621 tagcctttat cagaggccag tgggatggtg ggtgtaagag cttaaccaca actggtagtc
     1681 catagtgaag gcgaactgca ttctgggcca tctctgcttc ttggtgtcgg ctggtcagat
     1741 gacgaagagc acagatggca ggctcagtga tgtcttccct gtcaccagcc cgaaggacag
     1801 tacgcacaag agcctctata ccacccactt ggcagaccat catcttgttc ttataattat
     1861 tgcaagtgag gttagaaaga attccagctg cacaggtgac cacatttata tcatctgaac
     1921 ccagaagctg aacaagagtc ccaaggagac cttccatccc ttcctgttta gttgcagcat
     1981 ctgaaagatt cctgagagtc caaagacagt tctgaacaag acgttgactt ggatctgtca
     2041 ggtgaagtcc taaagcttgc attccaccag cttctacaat agccggctta ttactagagc
     2101 agacagatag caccttcagc actctgcttg tggtccacag tagtttttcg taagtatagg
     2161 tcctcattat atttactaaa gcttggggtc caccactagc cagtatgatg agcttgcttt
     2221 cttggttgcc ataagctaaa atttgaaggc agtctgtcgt aatagccaag aatttaacat
     2281 ttgttttgtt gagcaaggca accattttct gcagcccacc agctaaacgc actgccattt
     2341 tagctccttc ttgatgtaat aaaaggttgt ggagagttgt aatggcataa aacaacacag
     2401 aatccactgg tgaaccaagc attttcacca gggcaggaat gcctccagac ttaaagatgg
     2461 ccagtaagcc ctcacgatga tgggaaaggt tatgcaaggt cccagcggta caacgagctg
     2521 tttctacatc atttgtattc tgcatggtac gtacaatagc agacaccatc tgaggagaac
     2581 gcatgatagc gtgtctggaa gcttcctttt tagaaagctg atggaccata actgcagcct
     2641 tattaaccac cacctggtcc tcgtcattta gcagttttgt cagttcaggg attgcacgtg
     2701 tggcaagttc tgcatcatct tgatagttaa tcaagtttac aactgcatgt ttcagcatct
     2761 gtgatggttc agccaaacgc tggacattag tgggatgagc agcatcaaac tgtgtagatg
     2821 ggatctgcat gccctcatct aatgtctcag ggaacatagc agctcgtacc ctctgagctc
     2881 gagtcattgc atactgtcca tcaatatcag ctacttgttc ttgagtgaag gactgagaaa
     2941 atccctgttc ccactcatac aggacttggg aggtatccac atcctcttcc tcaggattgc
     3001 ctttaccact cagagaagga gctgtggtag tggcaccaga atggattcca gagtccaggt
     3061 aagactgttg ctgccagtga ctaacagccg cttttctgtc tggttccatg gccatgtcca
     3121 actccatcaa atcagcttga gtagccatgg cgatcgctag ctgcaaaaag aacaagtagc
     3181 ttgtattcta tagtgtcacc taaatgggcg atctgacggt tcactaaacc agctctgctt
     3241 atataggcct ccctggcgcg ccactagtta ttaatagtaa tcaattacgg ggtcattagt
     3301 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     3361 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     3421 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     3481 agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg
     3541 gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat
     3601 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg
     3661 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     3721 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc ccgccccgtt
     3781 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtcaaa tcgaattcta atactgttga
     3841 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     3901 ccaggaggca gatcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3961 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4021 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4081 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4141 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4201 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4261 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4321 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4381 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4441 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4501 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4561 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4621 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4681 tctatcgcct tcttgacgag ttcttctgat tcgaaatgac cgaccaagcg acgcccaacc
     4741 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4801 aaagaacatg tcaaacatga gaattacaac ttatatcgta tggggctgac ttcaggtgct
     4861 acatttgaag agataaattg cactgaaatc tagaaatatt ttatctgatt aataagatga
     4921 tcttcttgag atcgttttgg tctgcgcgta atctcttgct ctgaaaacga aaaaaccgcc
     4981 ttgcagggcg gtttttcgaa ggttctctga gctaccaact ctttgaaccg aggtaactgg
     5041 cttggaggag cgcagtcacc aaaacttgtc ctttcagttt agccttaacc ggcgcatgac
     5101 ttcaagacta actcctctaa atcaattacc agtggctgct gccagtggtg cttttgcatg
     5161 tctttccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cggactgaac
     5221 ggggggttcg tgcatacagt ccagcttgga gcgaactgcc tacccggaac tgagtgtcag
     5281 gcgtggaatg agacaaacgc ggccataaca gcggaatgac accggtaaac cgaaaggcag
     5341 gaacaggaga gcgcacgagg gagccgccag ggggaaacgc ctggtatctt tatagtcctg
     5401 tcgggtttcg ccaccactga tttgagcgtc agatttcgtg atgcttgtca ggggggcgga
     5461 gcctatggaa aaacggcttt gccgcggccc tctcacttcc ctgttaagta tcttcctggc
     5521 atcttccagg aaatctccgc cccgttcgta agccatttcc gctcgccgca gtcgaacgac
     5581 cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc ctgtatcaca tattctgctg
     5641 acgcaccggt gcagcctttt ttctcctgcc acatgaagca cttcactgac accctcatca
     5701 gtgccaacat agtaagccag tatacactcc gctagcgctg atgtccggaa tcgtcacaca
     5761 aaaaaccaac acacagatgt aatgaaaata aagatatttt attgcggcca tcgtgatggc
     5821 tagagtctaa actcgaccta cctccttagc gcgcggtcgt gtactgggcc ggcggttgcc
     5881 gctgtcggcc agcacctcga cgcagccgtc ggtccatacg gctgcgctcc ggcgagcgat
     5941 ttgtgtgcga ccgacggtgc cggccccgct gcggacgatg gcatcgcagc ggccttgagc
     6001 ccaggcagca tcgtcgaagt tgccgtccac gaggctctgg tagagctgat ccaggccgat
     6061 gcgcagcatg taggctcgca gacgagggct gccggccagc tcgggatgcc ggcgctcgaa
     6121 gtagcgagtc tgctgctcca tgcaagccag ccagggccgc cagaagaaga tgttggccac
     6181 ctcgtactga ctgtccccga acatagcttc ggaccagtcg attacggcgg tgatgcggcc
     6241 gttgtcggtc aggacgttgt tgctgccgaa gtcggcatgg acgaggtggc gcacctcggg
     6301 acagtcttcg gcccacagca tcagttcgtc cagggcttga gctacgctgg cggacacggt
     6361 gtcgtccatc acggtctgcc agtggtagac atggggatca gcaatggcgc aaatgaaatc
     6421 ccgccaggtt gtgtactggc cgatgccttg gggcccgaat gggccgaagc cgctggtttg
     6481 actgaggtcg gcggcggcaa tagcatccat agcttcggcg acaggctgta acacagcagg
     6541 cagctctgtt tcggggaggt cttggagagt aacgccttgg gcgcgtctac tgatgcagta
     6601 tgtcaggctc tcgctgaact cgccgatgtc caacacttcg gggatgggta gtgcagcgct
     6661 ggcgaagtgg cggtacacgt agcggtcttt gtagaagcca tcagcgcagc tattcacccg
     6721 cagtacatag ccgcgtccgc cgacatcgaa gctgaaggct cggctctctt cgccctccga
     6781 caactgcatc aggtcgctca cactgtcgaa cttctcgatg agaaattttt caacgctggt
     6841 agcggtgagt tcgggcttct tcatggtggc gctagtgtca gaagaatcga gctttttgca
     6901 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     6961 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     7021 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     7081 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
     7141 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     7201 ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc agaaggtacc
     7261 taaccaagtt cctctttcag aggttatttc aggccatggt gctgcgc
//
