LOCUS       sequence-471877-        7018 bp    DNA     circular SYN 24-JUN-2025
DEFINITION  Express beta catenin-HaloTag(R) 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 7018)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 7018)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoBRET Assay Vector, beta-catenin
FEATURES             Location/Qualifiers
     source          1..7018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1065..3407
                     /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             1065..3407
                     /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             3429..3449
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             3465..4349
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(4437..4455)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4492..4613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4503..4522)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4557..4576
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             5012..5806
                     /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="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5066..5085)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5676..5695
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5978..6566)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6058..6077)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6682..6965
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgat cgccatggct actcaagctg
     1081 atttgatgga gttggacatg gccatggaac cagacagaaa agcggctgtt agtcactggc
     1141 agcaacagtc ttacctggac tctggaatcc attctggtgc cactaccaca gctccttctc
     1201 tgagtggtaa aggcaatcct gaggaagagg atgtggatac ctcccaagtc ctgtatgagt
     1261 gggaacaggg attttctcag tccttcactc aagaacaagt agctgatatt gatggacagt
     1321 atgcaatgac tcgagctcag agggtacgag ctgctatgtt ccctgagaca ttagatgagg
     1381 gcatgcagat cccatctaca cagtttgatg ctgctcatcc cactaatgtc cagcgtttgg
     1441 ctgaaccatc acagatgctg aaacatgcag ttgtaaactt gattaactat caagatgatg
     1501 cagaacttgc cacacgtgca atccctgaac tgacaaaact gctaaatgac gaggaccagg
     1561 tggtggttaa taaggctgca gttatggtcc atcagctttc taaaaaggaa gcttccagac
     1621 acgctatcat gcgttctcct cagatggtgt ctgctattgt acgtaccatg cagaatacaa
     1681 atgatgtaga aacagctcgt tgtaccgctg ggaccttgca taacctttcc catcatcgtg
     1741 agggcttact ggccatcttt aagtctggag gcattcctgc cctggtgaaa atgcttggtt
     1801 caccagtgga ttctgtgttg ttttatgcca ttacaactct ccacaacctt ttattacatc
     1861 aagaaggagc taaaatggca gtgcgtttag ctggtgggct gcagaaaatg gttgccttgc
     1921 tcaacaaaac aaatgttaaa ttcttggcta ttacgacaga ctgccttcaa attttagctt
     1981 atggcaacca agaaagcaag ctcatcatac tggctagtgg tggaccccaa gctttagtaa
     2041 atataatgag gacctatact tacgaaaaac tactgtggac cacaagcaga gtgctgaagg
     2101 tgctatctgt ctgctctagt aataagccgg ctattgtaga agctggtgga atgcaagctt
     2161 taggacttca cctgacagat ccaagtcaac gtcttgttca gaactgtctt tggactctca
     2221 ggaatctttc agatgctgca actaaacagg aagggatgga aggtctcctt gggactcttg
     2281 ttcagcttct gggttcagat gatataaatg tggtcacctg tgcagctgga attctttcta
     2341 acctcacttg caataattat aagaacaaga tgatggtctg ccaagtgggt ggtatagagg
     2401 ctcttgtgcg tactgtcctt cgggctggtg acagggaaga catcactgag cctgccatct
     2461 gtgctcttcg tcatctgacc agccgacacc aagaagcaga gatggcccag aatgcagttc
     2521 gccttcacta tggactacca gttgtggtta agctcttaca cccaccatcc cactggcctc
     2581 tgataaaggc tactgttgga ttgattcgaa atcttgccct ttgtcccgca aatcatgcac
     2641 ctttgcgtga gcagggtgcc attccacgac tagttcagtt gcttgttcgt gcacatcagg
     2701 atacccagcg ccgtacgtcc atgggtggga cacagcagca atttgtggag ggggtccgca
     2761 tggaagaaat agttgaaggt tgtaccggag cccttcacat cctagctcgg gatgttcaca
     2821 accgaattgt tatcagagga ctaaatacca ttccattgtt tgtgcagctg ctttattctc
     2881 ccattgaaaa catccaaaga gtagctgcag gggtcctctg tgaacttgct caggacaagg
     2941 aagctgcaga agctattgaa gctgagggag ccacagctcc tctgacagag ttacttcact
     3001 ctaggaatga aggtgtggcg acatatgcag ctgctgtttt gttccgaatg tctgaggaca
     3061 agccacaaga ttacaagaaa cggctttcag ttgagctgac cagctctctc ttcagaacag
     3121 agccaatggc ttggaatgag actgctgatc ttggacttga tattggtgcc cagggagaac
     3181 cccttggata tcgccaggat gatcctagct atcgttcttt tcactctggt ggatatggcc
     3241 aggatgcctt gggtatggac cccatgatgg aacatgagat gggtggccac caccctggtg
     3301 ctgactatcc agttgatggg ctgccagatc tggggcatgc ccaggacctc atggatgggc
     3361 tgcctccagg tgacagcaat cagctggcct ggtttgatac tgacctggtt tctctcgagc
     3421 caaccactga ggatctgtac tttcagagcg ataacgatgg atccgaaatc ggtactggct
     3481 ttccattcga cccccattat gtggaagtcc tgggcgagcg catgcactac gtcgatgttg
     3541 gtccgcgcga tggcacccct gtgctgttcc tgcacggtaa cccgacctcc tcctacgtgt
     3601 ggcgcaacat catcccgcat gttgcaccga cccatcgctg cattgctcca gacctgatcg
     3661 gtatgggcaa atccgacaaa ccagacctgg gttatttctt cgacgaccac gtccgcttca
     3721 tggatgcctt catcgaagcc ctgggtctgg aagaggtcgt cctggtcatt cacgactggg
     3781 gctccgctct gggtttccac tgggccaagc gcaatccaga gcgcgtcaaa ggtattgcat
     3841 ttatggagtt catccgccct atcccgacct gggacgaatg gccagaattt gcccgcgaga
     3901 ccttccaggc cttccgcacc accgacgtcg gccgcaagct gatcatcgat cagaacgttt
     3961 ttatcgaggg tacgctgccg atgggtgtcg tccgcccgct gactgaagtc gagatggacc
     4021 attaccgcga gccgttcctg aatcctgttg accgcgagcc actgtggcgc ttcccaaacg
     4081 agctgccaat cgccggtgag ccagcgaaca tcgtcgcgct ggtcgaagaa tacatggact
     4141 ggctgcacca gtcccctgtc ccgaagctgc tgttctgggg caccccaggc gttctgatcc
     4201 caccggccga agccgctcgc ctggccaaaa gcctgcctaa ctgcaaggct gtggacatcg
     4261 gcccgggtct gaatctgctg caagaagaca acccggacct gatcggcagc gagatcgcgc
     4321 gctggctgtc tactctggag atttccggtt aatagaattc tagagtcgac ctgcaggcat
     4381 gcaagctgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     4441 tgagcaataa ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca
     4501 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     4561 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     4621 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca
     4681 agtaaaacct ctacaaatgt ggtaaaatcg aattctaatg gatcctcttt gcgcttgcgt
     4741 tttcccttgt ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg
     4801 gactggcttt ctacgtgttc cgcttccttt agcagccctt gcgccctgag tgcttgcggc
     4861 agcgtgagct tcaaaagaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg
     4921 caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc
     4981 tgatcaagag acaggatgac ggtcgtttcg catgcttgaa caagatggat tgcacgcagg
     5041 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5101 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5161 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     5221 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5281 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5341 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5401 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgcactc ggatggaagc
     5461 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5521 gttcgccagg ctcaaggcgc gtatgccgga tggtgaggat ctcgtcgtga ctcatggcga
     5581 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5641 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5701 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5761 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     5821 ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac
     5881 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5941 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6001 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6061 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6121 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6181 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6241 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6301 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6361 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     6421 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6481 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6541 ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg
     6601 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6661 tcacctagat ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg
     6721 ggatggtgaa accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg
     6781 taccctctgg ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt
     6841 ttttcgggtg gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt
     6901 ttagcggtgc gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc
     6961 atcgagataa accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
