LOCUS       sequence-482208-        5898 bp    DNA     circular SYN 06-AUG-2025
DEFINITION  Expresses V5-tagged and LgBiT-tagged B-catenin for use with the 
            Promega HiBiT system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5898)
  AUTHORS   Kassel S, Yuan K, Bunnag N, Neitzel LR, Lu W, Schwarzkopf A, Maines 
            B, Loberg MA, Xu G, Adams A, McCray AD, Cho A, Rockouski M, Orton G,
            Goldsmith L, Aronno MMA, Spencer ZT, Khan OM, Ye F, Williams C, 
            Lebensohn AM, Rohatgi R, Wang X, Weiss VL, Hong CC, Kettenbach AN, 
            Robbins DJ, Ahmed Y, Lee E
  TITLE     The TRIP12 E3 ligase induces SWI/SNF component BRG1-beta-catenin 
            interaction to promote Wnt signaling.
  JOURNAL   Nat Commun. 2025 Jun 5;16(1):5248. doi: 10.1038/s41467-025-60535-5.
  PUBMED    40473626
REFERENCE   2  (bases 1 to 5898)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2025-06-5"; volume: "16(1):5248. doi"; pages: " 
            10.1038/s41467-025-60535-5"
COMMENT     Sequence Label: pRP-B-catenin-V5-LgBiT
FEATURES             Location/Qualifiers
     source          1..5898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..55
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    81..101
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     enhancer        162..465
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        466..669
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     620..640
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    690..714
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      715..724
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             721..3063
                     /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             721..3063
                     /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"
     CDS             3112..3153
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(3682..3706)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3735..3856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3746..3765)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3800..3819
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3972..3989)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4143..4731)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4223..4242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4902..5693,5694..5762))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5525..5544
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5763..5867)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
       61 cgtcggcgcg ccgcggccgc caactttgta tagaaaagtt gtagttatta atagtaatca
      121 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      181 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      241 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      301 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      361 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      421 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      481 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      541 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      601 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      661 agcagagctg gtttagtgaa ccgtcagatc caagtttgta caaaaaagca ggctgccacc
      721 atggctactc aagctgattt gatggagttg gacatggcca tggaaccaga cagaaaagcg
      781 gctgttagtc actggcagca acagtcttac ctggactctg gaatccattc tggtgccact
      841 accacagctc cttctctgag tggtaaaggc aatcctgagg aagaggatgt ggatacctcc
      901 caagtcctgt atgagtggga acagggattt tctcagtcct tcactcaaga acaagtagct
      961 gatattgatg gacagtatgc aatgactcga gctcagaggg tacgagctgc tatgttccct
     1021 gagacattag atgagggcat gcagatccca tctacacagt ttgatgctgc tcatcccact
     1081 aatgtccagc gtttggctga accatcacag atgctgaaac atgcagttgt aaacttgatt
     1141 aactatcaag atgatgcaga acttgccaca cgtgcaatcc ctgaactgac aaaactgcta
     1201 aatgacgagg accaggtggt ggttaataag gctgcagtta tggtccatca gctttctaaa
     1261 aaggaagctt ccagacacgc tatcatgcgt tctcctcaga tggtgtctgc tattgtacgt
     1321 accatgcaga atacaaatga tgtagaaaca gctcgttgta ccgctgggac cttgcataac
     1381 ctttcccatc atcgtgaggg cttactggcc atctttaagt ctggaggcat tcctgccctg
     1441 gtgaaaatgc ttggttcacc agtggattct gtgttgtttt atgccattac aactctccac
     1501 aaccttttat tacatcaaga aggagctaaa atggcagtgc gtttagctgg tgggctgcag
     1561 aaaatggttg ccttgctcaa caaaacaaat gttaaattct tggctattac gacagactgc
     1621 cttcaaattt tagcttatgg caaccaagaa agcaagctca tcatactggc tagtggtgga
     1681 ccccaagctt tagtaaatat aatgaggacc tatacttacg aaaaactact gtggaccaca
     1741 agcagagtgc tgaaggtgct atctgtctgc tctagtaata agccggctat tgtagaagct
     1801 ggtggaatgc aagctttagg acttcacctg acagatccaa gtcaacgtct tgttcagaac
     1861 tgtctttgga ctctcaggaa tctttcagat gctgcaacta aacaggaagg gatggaaggt
     1921 ctccttggga ctcttgttca gcttctgggt tcagatgata taaatgtggt cacctgtgca
     1981 gctggaattc tttctaacct cacttgcaat aattataaga acaagatgat ggtctgccaa
     2041 gtgggtggta tagaggctct tgtgcgtact gtccttcggg ctggtgacag ggaagacatc
     2101 actgagcctg ccatctgtgc tcttcgtcat ctgaccagcc gacaccaaga agcagagatg
     2161 gcccagaatg cagttcgcct tcactatgga ctaccagttg tggttaagct cttacaccca
     2221 ccatcccact ggcctctgat aaaggctact gttggattga ttcgaaatct tgccctttgt
     2281 cccgcaaatc atgcaccttt gcgtgagcag ggtgccattc cacgactagt tcagttgctt
     2341 gttcgtgcac atcaggatac ccagcgccgt acgtccatgg gtgggacaca gcagcaattt
     2401 gtggaggggg tccgcatgga agaaatagtt gaaggttgta ccggagccct tcacatccta
     2461 gctcgggatg ttcacaaccg aattgttatc agaggactaa ataccattcc attgtttgtg
     2521 cagctgcttt attctcccat tgaaaacatc caaagagtag ctgcaggggt cctctgtgaa
     2581 cttgctcagg acaaggaagc tgcagaagct attgaagctg agggagccac agctcctctg
     2641 acagagttac ttcactctag gaatgaaggt gtggcgacat atgcagctgc tgttttgttc
     2701 cgaatgtctg aggacaagcc acaagattac aagaaacggc tttcagttga gctgaccagc
     2761 tctctcttca gaacagagcc aatggcttgg aatgagactg ctgatcttgg acttgatatt
     2821 ggtgcccagg gagaacccct tggatatcgc caggatgatc ctagctatcg ttcttttcac
     2881 tctggtggat atggccagga tgccttgggt atggacccca tgatggaaca tgagatgggt
     2941 ggccaccacc ctggtgctga ctatccagtt gatgggctgc cagatctggg gcatgcccag
     3001 gacctcatgg atgggctgcc tccaggtgac agcaatcagc tggcctggtt tgatactgac
     3061 ctgtctggtg gcggaggctc gggcggaggt gggtcgggtg gcggcggatc aggtaagcct
     3121 atccctaacc ctctcctcgg tctcgattct acgtctggtg gcggaggctc gggcggaggt
     3181 gggtcgggtg gcggcggatc aatggtgttc accctggagg acttcgtggg cgactgggag
     3241 cagaccgccg cctacaacct ggaccaggtg ctggagcagg gcggcgtgag cagcctgctg
     3301 cagaacctgg ccgtgagcgt gacccccatc cagagaatcg tgagaagcgg cgagaacgcc
     3361 ctgaagatcg acatccacgt gatcatcccc tacgagggcc tgagcgccga ccagatggcc
     3421 cagatcgagg aggtgttcaa ggtggtgtac cccgtggacg accaccactt caaggtgatc
     3481 ctgccctacg gcaccctggt gatcgacggc gtgaccccca acatgctgaa ctacttcggc
     3541 agaccctacg agggcatcgc cgtgttcgac ggcaagaaga tcaccgtgac cggcaccctg
     3601 tggaacggca acaagatcat cgacgagaga ctgatcaccc ccgacggcag catgctgttc
     3661 agagtgacca tcaacagcta aacccagctt tcttgtacaa agtggtgatg gccggccgct
     3721 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3781 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3841 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3901 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtagcg gccgcggcgc
     3961 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4021 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4081 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4141 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4201 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4261 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct ctcttcggga
     4321 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4381 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4441 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4501 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4561 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4621 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4681 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4741 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4801 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4861 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4921 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4981 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5041 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5101 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5161 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5221 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5281 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5341 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5401 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5461 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5521 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5581 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5641 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5701 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5761 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5821 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5881 aaagtgccac ctgacgtc
//
