LOCUS       sequence-485634-        8853 bp    DNA     circular SYN 09-SEP-2025
DEFINITION  Expression of human beta-catenin ∆arm 1-3 (deletion of a.a. 134-266)
            using piggyBac transposition.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8853)
  AUTHORS   Chan YS, Gao Q, Robinson SA, Wang W, Filandrova R, Weinhold LM, 
            Cabrera ML, Zhang M, Ambati CSR, Lerario AM, Putluri N, 
            Kiseljak-Vassiliades K, Wierman ME, Habra MA, Hammer GD, Veverka V, 
            Cermakova K, Hodges HC
  TITLE     beta-catenin functions as a molecular adapter for disordered cBAF 
            interactions.
  JOURNAL   Mol Cell. 2025 Jul 15:S1097-2765(25)00576-3. doi: 
            10.1016/j.molcel.2025.06.026.
  PUBMED    40695292
REFERENCE   2  (bases 1 to 8853)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 9, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell. 
            2025 Jul 15:S1097-2765(25)00576-3. doi: 
            10.1016/j.molcel.2025.06.026."
COMMENT     Sequence Label: pPB-CAG-CTNNB1-delta1-3-HA
FEATURES             Location/Qualifiers
     source          1..8853
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     343..362
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(581..685)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(711..1166)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(848..869)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1060..1079
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1293..1315
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1307..1324
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1308..1324
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1334..1353
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1334..1352
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   1382..1416
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        1737..2116
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3369..3391
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      3482..3491
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3533..3568
                     /codon_start=1
                     /product="nanobody epitope tag corresponding to amino acids
                     16-27 of human beta-catenin (Traenkle et al., 2015)"
                     /label=BC2 tag
                     /translation="PDRKAAVSHWQQ"
     CDS             5441..5530
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     misc_feature    5625..6174
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5769..5786)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5996..6015
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(6645..6662)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6651..6875
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6922..6938)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6946..6962
                     /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(6970..6976,6977..6994,6995..7000))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7015..7036
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   7270..7332
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7373..7393)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7373..7391)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7412..7428)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7436..7452
                     /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(7460..7466,7467..7484,7485..7490))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7505..7526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7643..7660)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7814..8402)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7894..7913)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8573..8853,1..511,512..580))
                     /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"
ORIGIN
        1 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
       61 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
      121 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
      181 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
      241 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
      301 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
      361 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
      421 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
      481 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
      541 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
      601 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
      661 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
      721 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
      781 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
      841 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
      901 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
      961 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     1021 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     1081 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     1141 gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga
     1201 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     1261 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     1321 cagtgagcgc gcgtaatacg actcactata gggcgaattg gggcgcgcca ttctagatta
     1381 accctagaaa gatagtctgc gtaaaattga cgcatgcatt cttgaaatat tgctctctct
     1441 ttctaaatag cgcgaatccg tcgctgtgca tttaggacat ctcagtcgcc gcttggagct
     1501 cccgtgaggc gtgcttgtca atgcggtaag tgtcactgat tttgaactat aacgaccgcg
     1561 tgagtcaaaa tgacgcatga ttatctttta cgtgactttt aagatttaac tcatacgata
     1621 attatattgt tatttcatgt tctacttacg tgataactta ttatatatat attttcttgt
     1681 tatagatatc aactagaatg ctagactcga gaagcttgat cggccacgcg gatctcgaca
     1741 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     1801 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1861 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     1921 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     1981 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     2041 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     2101 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
     2161 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg
     2221 ggcggggggg gggggggggg ggcccggggg gggggggggg gggggggggg ggggcgcgcg
     2281 gggggggggg gggggggggg aggggcgggg cggggcgagg cggagaggtg cggcggcagc
     2341 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc
     2401 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc
     2461 ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca
     2521 ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg
     2581 gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg
     2641 cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg
     2701 ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg
     2761 cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga
     2821 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
     2881 gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc
     2941 ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg
     3001 gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga
     3061 ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
     3121 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga
     3181 gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg
     3241 cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct
     3301 tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg
     3361 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agccggagcg cacgtcggca
     3421 gtcggctccc tcgttgaccg aatcaccgac ctctctcccc agggggatcc accgggctag
     3481 cgccaccatg gctactcaag ctgatttgat ggagttggac atggccatgg aaccagacag
     3541 aaaagcggct gttagtcact ggcagcaaca gtcttacctg gactctggaa tccattctgg
     3601 tgccactacc acagctcctt ctctgagtgg taaaggcaat cctgaggaag aggatgtgga
     3661 tacctcccaa gtcctgtatg agtgggaaca gggattttct cagtccttca ctcaagaaca
     3721 agtagctgat attgatggac agtatgcaat gactcgagct cagagggtac gagctgctat
     3781 gttccctgag acattagatg agggcatgca gatcccatct acacagtttg atgctgctca
     3841 tcccactaat gtccagcgtt tggctgaacc atcacagatg ctgaaagaag gagctaaaat
     3901 ggcagtgcgt ttagctggtg ggctgcagaa aatggttgcc ttgctcaaca aaacaaatgt
     3961 taaattcttg gctattacga cagactgcct tcaaatttta gcttatggca accaagaaag
     4021 caagctcatc atactggcta gtggtggacc ccaagcttta gtaaatataa tgaggaccta
     4081 tacttacgaa aaactactgt ggaccacaag cagagtgctg aaggtgctat ctgtctgctc
     4141 tagtaataag ccggctattg tagaagctgg tggaatgcaa gctttaggac ttcacctgac
     4201 agatccaagt caacgtcttg ttcagaactg tctttggact ctcaggaatc tttcagatgc
     4261 tgcaactaaa caggaaggga tggaaggtct ccttgggact cttgttcagc ttctgggttc
     4321 agatgatata aatgtggtca cctgtgcagc tggaattctt tctaacctca cttgcaataa
     4381 ttataagaac aagatgatgg tctgccaagt gggtggtata gaggctcttg tgcgtactgt
     4441 ccttcgggct ggtgacaggg aagacatcac tgagcctgcc atctgtgctc ttcgtcatct
     4501 gaccagccga caccaagaag cagagatggc ccagaatgca gttcgccttc actatggact
     4561 accagttgtg gttaagctct tacacccacc atcccactgg cctctgataa aggctactgt
     4621 tggattgatt cgaaatcttg ccctttgtcc cgcaaatcat gcacctttgc gtgagcaggg
     4681 tgccattcca cgactagttc agttgcttgt tcgtgcacat caggataccc agcgccgtac
     4741 gtccatgggt gggacacagc agcaatttgt ggagggggtc cgcatggaag aaatagttga
     4801 aggttgtacc ggagcccttc acatcctagc tcgggatgtt cacaaccgaa ttgttatcag
     4861 aggactaaat accattccat tgtttgtgca gctgctttat tctcccattg aaaacatcca
     4921 aagagtagct gcaggggtcc tctgtgaact tgctcaggac aaggaagctg cagaagctat
     4981 tgaagctgag ggagccacag ctcctctgac agagttactt cactctagga atgaaggtgt
     5041 ggcgacatat gcagctgctg ttttgttccg aatgtctgag gacaagccac aagattacaa
     5101 gaaacggctt tcagttgagc tgaccagctc tctcttcaga acagagccaa tggcttggaa
     5161 tgagactgct gatcttggac ttgatattgg tgcccaggga gaaccccttg gatatcgcca
     5221 ggatgatcct agctatcgtt cttttcactc tggtggatat ggccaggatg ccttgggtat
     5281 ggaccccatg atggaacatg agatgggtgg ccaccaccct ggtgctgact atccagttga
     5341 tgggctgcca gatctggggc atgcccagga cctcatggat gggctgcctc caggtgacag
     5401 caatcagctg gcctggtttg atactgacct gaccggtatg tacccatacg atgttccaga
     5461 ttacgctggc tatccctatg acgtcccgga ctatgcaggc tcctatccat atgacgttcc
     5521 agattacgct tgaacgcgtt agtgcggatc ctagtgcgaa ttctagtgcg tcgactctag
     5581 caacggtttc cctctagcgg gatcaattcc gccccccccc cctaacgtta ctggccgaag
     5641 ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc
     5701 ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg
     5761 tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc
     5821 tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc
     5881 cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa
     5941 ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct
     6001 ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg
     6061 atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg
     6121 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat aataccatgg
     6181 tcatgaaaac atttaacatt tctcaacaag atctagaatt agtagaagta gcgacagaga
     6241 agattacaat gctttatgag gataataaac atcatgtggg agcggcaatt cgtacgaaaa
     6301 caggagaaat catttcggca gtacatattg aagcgtatat aggacgagta actgtttgtg
     6361 cagaagccat tgcgattggt agtgcagttt cgaatggaca aaaggatttt gacacgattg
     6421 tagctgttag acacccttat tctgacgaag tagatagaag tattcgagtg gtaagtcctt
     6481 gtggtatgtg tagggagttg atttcagact atgcaccaga ttgttttgtg ttaatagaaa
     6541 tgaatggcaa gttagtcaaa actacgattg aagaactcat tccactcaaa tatacccgaa
     6601 attaaaagtc gagatgcatc gatgatctag agctcgctga tcagcctcga ctgtgccttc
     6661 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     6721 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     6781 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     6841 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaacctgcag
     6901 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6961 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     7021 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     7081 gtgccagcgg atcttaaaag ttttgttact ttatagaaga aattttgagt ttttgttttt
     7141 ttttaataaa taaataaaca taaataaatt gtttgttgaa tttattatta gtatgtaagt
     7201 gtaaatataa taaaacttaa tatctattca aattaataaa taaacctcga tatacagacc
     7261 gataaaacac atgcgtcaat tttacgcatg attatcttta acgtacgtca caatatgatt
     7321 atctttctag ggttaatcta gtatcgtaga ttaattaatc cagcttttgt tccctttagt
     7381 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     7441 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     7501 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     7561 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     7621 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     7681 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     7741 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     7801 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     7861 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7921 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7981 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     8041 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     8101 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     8161 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     8221 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     8281 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     8341 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     8401 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     8461 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     8521 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     8581 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     8641 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     8701 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     8761 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     8821 attgttgccg ggaagctaga gtaagtagtt cgc
//
