LOCUS       sequence-471916-        7439 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express LgBiT-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 7439)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 7439)
  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: LgBiT-Beta catenin Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7439
                     /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"
     primer_bind     275..293
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(401..2749)
                     /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"
     promoter        complement(3319..3337)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3320..3337)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        3457..3760
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3915..3935
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3968..4015
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4047..4841
                     /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(4101..4120)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4711..4730
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      5058..5603
                     /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     5504..5523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5720..5737
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    5887..5935
                     /note="synthetic polyadenylation signal"
     CDS             complement(5959..6996)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5972..5980
                     /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(7027..7384)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7041..7176
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        7193..7384
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7341..7361
                     /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 ccccaagggg ttatgctagt tattgctcag cggtggcagc
      301 agccaactca gcttcctttc gggctttgtt agcagccgga tcagcttgca tgcctgcagg
      361 tcgactctag aggatccccg ggtaccgagc ccgaattcgt ttaaaccagg tcagtatcaa
      421 accaggccag ctgattgctg tcacctggag gcagcccatc catgaggtcc tgggcatgcc
      481 ccagatctgg cagcccatca actggatagt cagcaccagg gtggtggcca cccatctcat
      541 gttccatcat ggggtccata cccaaggcat cctggccata tccaccagag tgaaaagaac
      601 gatagctagg atcatcctgg cgatatccaa ggggttctcc ctgggcacca atatcaagtc
      661 caagatcagc agtctcattc caagccattg gctctgttct gaagagagag ctggtcagct
      721 caactgaaag ccgtttcttg taatcttgtg gcttgtcctc agacattcgg aacaaaacag
      781 cagctgcata tgtcgccaca ccttcattcc tagagtgaag taactctgtc agaggagctg
      841 tggctccctc agcttcaata gcttctgcag cttccttgtc ctgagcaagt tcacagagga
      901 cccctgcagc tactctttgg atgttttcaa tgggagaata aagcagctgc acaaacaatg
      961 gaatggtatt tagtcctctg ataacaattc ggttgtgaac atcccgagct aggatgtgaa
     1021 gggctccggt acaaccttca actatttctt ccatgcggac cccctccaca aattgctgct
     1081 gtgtcccacc catggacgta cggcgctggg tatcctgatg tgcacgaaca agcaactgaa
     1141 ctagtcgtgg aatggcaccc tgctcacgca aaggtgcatg atttgcggga caaagggcaa
     1201 gatttcgaat caatccaaca gtagccttta tcagaggcca gtgggatggt gggtgtaaga
     1261 gcttaaccac aactggtagt ccatagtgaa ggcgaactgc attctgggcc atctctgctt
     1321 cttggtgtcg gctggtcaga tgacgaagag cacagatggc aggctcagtg atgtcttccc
     1381 tgtcaccagc ccgaaggaca gtacgcacaa gagcctctat accacccact tggcagacca
     1441 tcatcttgtt cttataatta ttgcaagtga ggttagaaag aattccagct gcacaggtga
     1501 ccacatttat atcatctgaa cccagaagct gaacaagagt cccaaggaga ccttccatcc
     1561 cttcctgttt agttgcagca tctgaaagat tcctgagagt ccaaagacag ttctgaacaa
     1621 gacgttgact tggatctgtc aggtgaagtc ctaaagcttg cattccacca gcttctacaa
     1681 tagccggctt attactagag cagacagata gcaccttcag cactctgctt gtggtccaca
     1741 gtagtttttc gtaagtatag gtcctcatta tatttactaa agcttggggt ccaccactag
     1801 ccagtatgat gagcttgctt tcttggttgc cataagctaa aatttgaagg cagtctgtcg
     1861 taatagccaa gaatttaaca tttgttttgt tgagcaaggc aaccattttc tgcagcccac
     1921 cagctaaacg cactgccatt ttagctcctt cttgatgtaa taaaaggttg tggagagttg
     1981 taatggcata aaacaacaca gaatccactg gtgaaccaag cattttcacc agggcaggaa
     2041 tgcctccaga cttaaagatg gccagtaagc cctcacgatg atgggaaagg ttatgcaagg
     2101 tcccagcggt acaacgagct gtttctacat catttgtatt ctgcatggta cgtacaatag
     2161 cagacaccat ctgaggagaa cgcatgatag cgtgtctgga agcttccttt ttagaaagct
     2221 gatggaccat aactgcagcc ttattaacca ccacctggtc ctcgtcattt agcagttttg
     2281 tcagttcagg gattgcacgt gtggcaagtt ctgcatcatc ttgatagtta atcaagttta
     2341 caactgcatg tttcagcatc tgtgatggtt cagccaaacg ctggacatta gtgggatgag
     2401 cagcatcaaa ctgtgtagat gggatctgca tgccctcatc taatgtctca gggaacatag
     2461 cagctcgtac cctctgagct cgagtcattg catactgtcc atcaatatca gctacttgtt
     2521 cttgagtgaa ggactgagaa aatccctgtt cccactcata caggacttgg gaggtatcca
     2581 catcctcttc ctcaggattg cctttaccac tcagagaagg agctgtggta gtggcaccag
     2641 aatggattcc agagtccagg taagactgtt gctgccagtg actaacagcc gcttttctgt
     2701 ctggttccat ggccatgtcc aactccatca aatcagcttg agtagccatg gcgatcgcac
     2761 cgctcgagcc tccacctccg ctcccgccac caccggagct cccactgttg atggttactc
     2821 ggaacagcat ggagccgtcg ggggtgatca ggcgctcgtc gataattttg ttgccgttcc
     2881 acagggtccc tgttacagtg atctttttgc cgtcgaacac ggcgatgcct tcatacggcc
     2941 gtccgaaata gttcagcatg ttcggcgtaa ccccgtcgat taccagtgtg ccatagggca
     3001 ggatcacctt aaagtgatga tcatccacag ggtacaccac cttaaacacc tcttcgatct
     3061 gggccatttg gtcggcgctc agaccttcat acgggatgat gacatggatg tcgatcttca
     3121 gggcattttc accgctccgg acaatccttt ggatcggagt tacggacacg gcgagattct
     3181 gcagcaaact ggacacacct ccctgttcaa ggacttggtc caggttgtag gcggctgtct
     3241 gttcccagtc cccaacgaaa tcttcgagtg tgaagaccat ggtgagcgct agctgcaaaa
     3301 agaacaagta gcttgtattc tatagtgtca cctaaatggg cgatctgacg gttcactaaa
     3361 ccagctctgc ttatataggc ctccctggcg cgccactagt tattaatagt aatcaattac
     3421 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     3481 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     3541 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     3601 tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa
     3661 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac
     3721 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     3781 caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     3841 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa
     3901 ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtca aatcgaattc
     3961 taatactgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     4021 gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac gcaggttctc
     4081 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4141 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4201 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4261 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4321 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4381 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4441 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4501 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4561 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4621 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4681 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4741 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4801 agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg accgaccaag
     4861 cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4921 ggataacgca ggaaagaaca tgtcaaacat gagaattaca acttatatcg tatggggctg
     4981 acttcaggtg ctacatttga agagataaat tgcactgaaa tctagaaata ttttatctga
     5041 ttaataagat gatcttcttg agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac
     5101 gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct gagctaccaa ctctttgaac
     5161 cgaggtaact ggcttggagg agcgcagtca ccaaaacttg tcctttcagt ttagccttaa
     5221 ccggcgcatg acttcaagac taactcctct aaatcaatta ccagtggctg ctgccagtgg
     5281 tgcttttgca tgtctttccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5341 gtcggactga acggggggtt cgtgcataca gtccagcttg gagcgaactg cctacccgga
     5401 actgagtgtc aggcgtggaa tgagacaaac gcggccataa cagcggaatg acaccggtaa
     5461 accgaaaggc aggaacagga gagcgcacga gggagccgcc agggggaaac gcctggtatc
     5521 tttatagtcc tgtcgggttt cgccaccact gatttgagcg tcagatttcg tgatgcttgt
     5581 caggggggcg gagcctatgg aaaaacggct ttgccgcggc cctctcactt ccctgttaag
     5641 tatcttcctg gcatcttcca ggaaatctcc gccccgttcg taagccattt ccgctcgccg
     5701 cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa gcggaatata tcctgtatca
     5761 catattctgc tgacgcaccg gtgcagcctt ttttctcctg ccacatgaag cacttcactg
     5821 acaccctcat cagtgccaac atagtaagcc agtatacact ccgctagcgc tgatgtccgg
     5881 aatcgtcaca caaaaaacca acacacagat gtaatgaaaa taaagatatt ttattgcggc
     5941 catcgtgatg gctagagtct aaactcgacc tacctcctta gcgcgcggtc gtgtactggg
     6001 ccggcggttg ccgctgtcgg ccagcacctc gacgcagccg tcggtccata cggctgcgct
     6061 ccggcgagcg atttgtgtgc gaccgacggt gccggccccg ctgcggacga tggcatcgca
     6121 gcggccttga gcccaggcag catcgtcgaa gttgccgtcc acgaggctct ggtagagctg
     6181 atccaggccg atgcgcagca tgtaggctcg cagacgaggg ctgccggcca gctcgggatg
     6241 ccggcgctcg aagtagcgag tctgctgctc catgcaagcc agccagggcc gccagaagaa
     6301 gatgttggcc acctcgtact gactgtcccc gaacatagct tcggaccagt cgattacggc
     6361 ggtgatgcgg ccgttgtcgg tcaggacgtt gttgctgccg aagtcggcat ggacgaggtg
     6421 gcgcacctcg ggacagtctt cggcccacag catcagttcg tccagggctt gagctacgct
     6481 ggcggacacg gtgtcgtcca tcacggtctg ccagtggtag acatggggat cagcaatggc
     6541 gcaaatgaaa tcccgccagg ttgtgtactg gccgatgcct tggggcccga atgggccgaa
     6601 gccgctggtt tgactgaggt cggcggcggc aatagcatcc atagcttcgg cgacaggctg
     6661 taacacagca ggcagctctg tttcggggag gtcttggaga gtaacgcctt gggcgcgtct
     6721 actgatgcag tatgtcaggc tctcgctgaa ctcgccgatg tccaacactt cggggatggg
     6781 tagtgcagcg ctggcgaagt ggcggtacac gtagcggtct ttgtagaagc catcagcgca
     6841 gctattcacc cgcagtacat agccgcgtcc gccgacatcg aagctgaagg ctcggctctc
     6901 ttcgccctcc gacaactgca tcaggtcgct cacactgtcg aacttctcga tgagaaattt
     6961 ttcaacgctg gtagcggtga gttcgggctt cttcatggtg gcgctagtgt cagaagaatc
     7021 gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     7081 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     7141 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     7201 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     7261 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     7321 ggagcctggg gactttccac accctaactg acacacattc cacagctggt tctttccgcc
     7381 tcagaaggta cctaaccaag ttcctctttc agaggttatt tcaggccatg gtgctgcgc
//
