LOCUS       sequence-291737-        5435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For making Beta-catenin-S33Y RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5435)
  AUTHORS   Gambini A, Stein P, Savy V, Grow EJ, Papas BN, Zhang Y, Kenan AC, 
            Padilla-Banks E, Cairns BR, Williams CJ
  TITLE     Developmentally Programmed Tankyrase Activity Upregulates 
            beta-Catenin and Licenses Progression of Embryonic Genome 
            Activation.
  JOURNAL   Dev Cell. 2020 Jun 8;53(5):545-560.e7. doi: 
            10.1016/j.devcel.2020.04.018. Epub 2020 May 21.
  PUBMED    32442396
REFERENCE   2  (bases 1 to 5435)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.devcel.2020.04.018"; journalName: "Dev Cell"; date: 
            "2020-06-8- 8"; volume: "53"; issue: "5"; pages: "545-560.e7"
COMMENT     Sequence Label: pGEMHE-NLS-Beta-catenin-S33Y
FEATURES             Location/Qualifiers
     source          1..5435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           51..93
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             135..155
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             159..179
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             180..2519
                     /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="ATQADLMELDMAMEPDRKAAVSHWQQQSYLDYGIHSGATTTAPSL
                     SGKGNPEEEDVDTSQVLYEWEQGFSQSFTQEQVADIDGQYAMTRAQRVRAAMFPETLDE
                     GMQIPSTQFDAAHPTNVQRLAEPSQMLKHAVVNLINYQDDAELATRAIPELTKLLNDED
                     QVVVNKAAVMVHQLSKKEASRHAIMRSPQMVSAIVRTMQNTNDVETARCTAGTLHNLSH
                     HREGLLAIFKSGGIPALVKMLGSPVDSVLFYAITTLHNLLLHQEGAKMAVRLAGGLQKM
                     VALLNKTNVKFLAITTDCLQILAYGNQESKLIILASGGPQALVNIMRTYTYEKLLWTTS
                     RVLKVLSVCSSNKPAIVEAGGMQALGLHLTDPSQRLVQNCLWTLRNLSDAATKQEGMEG
                     LLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRTVLRAGDRED
                     ITEPAICALRHLTSRHQEAEMAQNAVRLHYGLPVVVKLLHPPSHWPLIKATVGLIRNLA
                     LCPANHAPLREQGAIPRLVQLLVRAHQDTQRRTSMGGTQQQFVEGVRMEEIVEGCTGAL
                     HILARDVHNRIVIRGLNTIPLFVQLLYSPIENIQRVAAGVLCELAQDKEAAEAIEAEGA
                     TAPLTELLHSRNEGVATYAAAVLFRMSEDKPQDYKKRLSVELTSSLFRTEPMAWNETAD
                     LGLDIGAQGEPLGYRQDDPSYRSFHSGGYGQDALGMDPMMEHEMGGHHPGADYPVDGLP
                     DLGHAQDLMDGLPPGDSNQLAWFDTDL"
     CDS             180..2519
                     /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="ATQADLMELDMAMEPDRKAAVSHWQQQSYLDYGIHSGATTTAPSL
                     SGKGNPEEEDVDTSQVLYEWEQGFSQSFTQEQVADIDGQYAMTRAQRVRAAMFPETLDE
                     GMQIPSTQFDAAHPTNVQRLAEPSQMLKHAVVNLINYQDDAELATRAIPELTKLLNDED
                     QVVVNKAAVMVHQLSKKEASRHAIMRSPQMVSAIVRTMQNTNDVETARCTAGTLHNLSH
                     HREGLLAIFKSGGIPALVKMLGSPVDSVLFYAITTLHNLLLHQEGAKMAVRLAGGLQKM
                     VALLNKTNVKFLAITTDCLQILAYGNQESKLIILASGGPQALVNIMRTYTYEKLLWTTS
                     RVLKVLSVCSSNKPAIVEAGGMQALGLHLTDPSQRLVQNCLWTLRNLSDAATKQEGMEG
                     LLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRTVLRAGDRED
                     ITEPAICALRHLTSRHQEAEMAQNAVRLHYGLPVVVKLLHPPSHWPLIKATVGLIRNLA
                     LCPANHAPLREQGAIPRLVQLLVRAHQDTQRRTSMGGTQQQFVEGVRMEEIVEGCTGAL
                     HILARDVHNRIVIRGLNTIPLFVQLLYSPIENIQRVAAGVLCELAQDKEAAEAIEAEGA
                     TAPLTELLHSRNEGVATYAAAVLFRMSEDKPQDYKKRLSVELTSSLFRTEPMAWNETAD
                     LGLDIGAQGEPLGYRQDDPSYRSFHSGGYGQDALGMDPMMEHEMGGHHPGADYPVDGLP
                     DLGHAQDLMDGLPPGDSNQLAWFDTDL"
     CDS             2520..2567
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     promoter        complement(2777..2795)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2778..2795)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2813..2829)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2813..2829)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2826..2848)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2837..2853
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2861..2867,2868..2885,2886..2891))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2906..2927
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3044..3061)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3215..3803)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3295..3314)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3974..4765,4766..4834))
                     /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     4597..4616
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4835..4939)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5007..5025
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5063..5085)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5185..5204
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5398..5420
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5412..5429
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5413..5429
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattaatt cgagctcggt acccagcttg cttgttcttt
       61 ttgcagaagc tcagaataaa cgctcaactt tggcagatca attccccggt gctagcgcta
      121 ccggtgccac catgccaaag aagaagagaa aggtagatcc aaaaaagaag agaaaggtag
      181 ctactcaagc tgatttgatg gagttggaca tggccatgga accagacaga aaagcggctg
      241 ttagtcactg gcagcaacag tcttacctgg actatggaat ccattctggt gccactacca
      301 cagctccttc tctgagtggt aaaggcaatc ctgaggaaga ggatgtggat acctcccaag
      361 tcctgtatga gtgggaacag ggattttctc agtccttcac tcaagaacaa gtagctgata
      421 ttgatggaca gtatgcaatg actcgagctc agagggtacg agctgctatg ttccctgaga
      481 cattagatga gggcatgcag atcccatcta cacagtttga tgctgctcat cccactaatg
      541 tccagcgttt ggctgaacca tcacagatgc tgaaacatgc agttgtaaac ttgattaact
      601 atcaagatga tgcagaactt gccacacgtg caatccctga actgacaaaa ctgctaaatg
      661 acgaggacca ggtggtggtt aataaggctg cagttatggt ccatcagctt tctaaaaagg
      721 aagcttccag acacgctatc atgcgttctc ctcagatggt gtctgctatt gtacgtacca
      781 tgcagaatac aaatgatgta gaaacagctc gttgtaccgc tgggaccttg cataaccttt
      841 cccatcatcg tgagggctta ctggccatct ttaagtctgg aggcattcct gccctggtga
      901 aaatgcttgg ttcaccagtg gattctgtgt tgttttatgc cattacaact ctccacaacc
      961 ttttattaca tcaagaagga gctaaaatgg cagtgcgttt agctggtggg ctgcagaaaa
     1021 tggttgcctt gctcaacaaa acaaatgtta aattcttggc tattacgaca gactgccttc
     1081 aaattttagc ttatggcaac caagaaagca agctcatcat actggctagt ggtggacccc
     1141 aagctttagt aaatataatg aggacctata cttacgaaaa actactgtgg accacaagca
     1201 gagtgctgaa ggtgctatct gtctgctcta gtaataagcc ggctattgta gaagctggtg
     1261 gaatgcaagc tttaggactt cacctgacag atccaagtca acgtcttgtt cagaactgtc
     1321 tttggactct caggaatctt tcagatgctg caactaaaca ggaagggatg gaaggtctcc
     1381 ttgggactct tgttcagctt ctgggttcag atgatataaa tgtggtcacc tgtgcagctg
     1441 gaattctttc taacctcact tgcaataatt ataagaacaa gatgatggtc tgccaagtgg
     1501 gtggtataga ggctcttgtg cgtactgtcc ttcgggctgg tgacagggaa gacatcactg
     1561 agcctgccat ctgtgctctt cgtcatctga ccagccgaca ccaagaagca gagatggccc
     1621 agaatgcagt tcgccttcac tatggactac cagttgtggt taagctctta cacccaccat
     1681 cccactggcc tctgataaag gctactgttg gattgattcg aaatcttgcc ctttgtcccg
     1741 caaatcatgc acctttgcgt gagcagggtg ccattccacg actagttcag ttgcttgttc
     1801 gtgcacatca ggatacccag cgccgtacgt ccatgggtgg gacacagcag caatttgtgg
     1861 agggggtccg catggaagaa atagttgaag gttgtaccgg agcccttcac atcctagctc
     1921 gggatgttca caaccgaatt gttatcagag gactaaatac cattccattg tttgtgcagc
     1981 tgctttattc tcccattgaa aacatccaaa gagtagctgc aggggtcctc tgtgaacttg
     2041 ctcaggacaa ggaagctgca gaagctattg aagctgaggg agccacagct cctctgacag
     2101 agttacttca ctctaggaat gaaggtgtgg cgacatatgc agctgctgtt ttgttccgaa
     2161 tgtctgagga caagccacaa gattacaaga aacggctttc agttgagctg accagctctc
     2221 tcttcagaac agagccaatg gcttggaatg agactgctga tcttggactt gatattggtg
     2281 cccagggaga accccttgga tatcgccagg atgatcctag ctatcgttct tttcactctg
     2341 gtggatatgg ccaggatgcc ttgggtatgg accccatgat ggaacatgag atgggtggcc
     2401 accaccctgg tgctgactat ccagttgatg ggctgccaga tctggggcat gcccaggacc
     2461 tcatggatgg gctgcctcca ggtgacagca atcagctggc ctggtttgat actgacctgg
     2521 actacaagga cgacgatgac aaggactaca aagacgatga cgacaagtaa tctagataaa
     2581 tgatagcttg atctggttac cactaaacca gcctcaagaa cacccgaatg gagtctctaa
     2641 gctacataat accaacttac actttacaaa atgttgtccc ccaaaatgta gccattcgta
     2701 tctgctccta ataaaaagaa msyycccctg caggcgcgcc gccggcgatt taaattaatt
     2761 aatctagctt gagtattcta tagtgtcacc taaatagctt ggcgtaatca tggtcatagc
     2821 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     2881 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     2941 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3001 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3061 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3121 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3181 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3241 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3301 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3361 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3421 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3481 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3541 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3601 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3661 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3721 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3781 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3841 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3901 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3961 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4021 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4081 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4141 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4201 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4261 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4321 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4381 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4441 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4501 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4561 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4621 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4681 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4741 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4801 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4861 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4921 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     4981 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     5041 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     5101 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     5161 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     5221 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     5281 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     5341 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     5401 agtcacgacg ttgtaaaacg acggccagtg aattg
//
