LOCUS       sequence-485618-        8786 bp    DNA     circular SYN 09-SEP-2025
DEFINITION  Expression of human beta-catenin ∆arm 10-12 (deletion of a.a. 
            522-664) using piggyBac transposition.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8786)
  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 8786)
  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-delta10-12-HA
FEATURES             Location/Qualifiers
     source          1..8786
                     /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"
     promoter        2118..2391
                     /label=chicken beta-actin promoter
     primer_bind     3332..3354
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      3445..3454
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3496..3531
                     /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             5374..5463
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     misc_feature    5558..6107
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5702..5719)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5929..5948
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(6578..6595)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6584..6808
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6855..6871)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6879..6895
                     /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(6903..6909,6910..6927,6928..6933))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6948..6969
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   7203..7265
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7306..7326)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7306..7324)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7345..7361)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7369..7385
                     /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(7393..7399,7400..7417,7418..7423))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7438..7459
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7576..7593)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7747..8335)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7827..7846)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8506..8786,1..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 gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg
     2281 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
     2341 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
     2401 gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
     2461 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
     2521 aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag
     2581 gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg
     2641 tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc
     2701 ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc
     2761 ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg
     2821 gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc
     2881 cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg
     2941 ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct
     3001 cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga
     3061 ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt
     3121 cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc
     3181 gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
     3241 gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac
     3301 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
     3361 tctagccgga gcgcacgtcg gcagtcggct ccctcgttga ccgaatcacc gacctctctc
     3421 cccaggggga tccaccgggc tagcgccacc atggctactc aagctgattt gatggagttg
     3481 gacatggcca tggaaccaga cagaaaagcg gctgttagtc actggcagca acagtcttac
     3541 ctggactctg gaatccattc tggtgccact accacagctc cttctctgag tggtaaaggc
     3601 aatcctgagg aagaggatgt ggatacctcc caagtcctgt atgagtggga acagggattt
     3661 tctcagtcct tcactcaaga acaagtagct gatattgatg gacagtatgc aatgactcga
     3721 gctcagaggg tacgagctgc tatgttccct gagacattag atgagggcat gcagatccca
     3781 tctacacagt ttgatgctgc tcatcccact aatgtccagc gtttggctga accatcacag
     3841 atgctgaaac atgcagttgt aaacttgatt aactatcaag atgatgcaga acttgccaca
     3901 cgtgcaatcc ctgaactgac aaaactgcta aatgacgagg accaggtggt ggttaataag
     3961 gctgcagtta tggtccatca gctttctaaa aaggaagctt ccagacacgc tatcatgcgt
     4021 tctcctcaga tggtgtctgc tattgtacgt accatgcaga atacaaatga tgtagaaaca
     4081 gctcgttgta ccgctgggac cttgcataac ctttcccatc atcgtgaggg cttactggcc
     4141 atctttaagt ctggaggcat tcctgccctg gtgaaaatgc ttggttcacc agtggattct
     4201 gtgttgtttt atgccattac aactctccac aaccttttat tacatcaaga aggagctaaa
     4261 atggcagtgc gtttagctgg tgggctgcag aaaatggttg ccttgctcaa caaaacaaat
     4321 gttaaattct tggctattac gacagactgc cttcaaattt tagcttatgg caaccaagaa
     4381 agcaagctca tcatactggc tagtggtgga ccccaagctt tagtaaatat aatgaggacc
     4441 tatacttacg aaaaactact gtggaccaca agcagagtgc tgaaggtgct atctgtctgc
     4501 tctagtaata agccggctat tgtagaagct ggtggaatgc aagctttagg acttcacctg
     4561 acagatccaa gtcaacgtct tgttcagaac tgtctttgga ctctcaggaa tctttcagat
     4621 gctgcaacta aacaggaagg gatggaaggt ctccttggga ctcttgttca gcttctgggt
     4681 tcagatgata taaatgtggt cacctgtgca gctggaattc tttctaacct cacttgcaat
     4741 aattataaga acaagatgat ggtctgccaa gtgggtggta tagaggctct tgtgcgtact
     4801 gtccttcggg ctggtgacag ggaagacatc actgagcctg ccatctgtgc tcttcgtcat
     4861 ctgaccagcc gacaccaaga agcagagatg gcccagaatg cagttcgcct tcactatgga
     4921 ctaccagttg tggttaagct cttacaccca ccatcccact ggcctctgat aaaggctact
     4981 gttggattga ttcgaaatct tgccctttgt cccgacaagc cacaagatta caagaaacgg
     5041 ctttcagttg agctgaccag ctctctcttc agaacagagc caatggcttg gaatgagact
     5101 gctgatcttg gacttgatat tggtgcccag ggagaacccc ttggatatcg ccaggatgat
     5161 cctagctatc gttcttttca ctctggtgga tatggccagg atgccttggg tatggacccc
     5221 atgatggaac atgagatggg tggccaccac cctggtgctg actatccagt tgatgggctg
     5281 ccagatctgg ggcatgccca ggacctcatg gatgggctgc ctccaggtga cagcaatcag
     5341 ctggcctggt ttgatactga cctgaccggt atgtacccat acgatgttcc agattacgct
     5401 ggctatccct atgacgtccc ggactatgca ggctcctatc catatgacgt tccagattac
     5461 gcttgaacgc gttagtgcgg atcctagtgc gaattctagt gcgtcgactc tagcaacggt
     5521 ttccctctag cgggatcaat tccgcccccc ccccctaacg ttactggccg aagccgcttg
     5581 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc
     5641 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc
     5701 cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa
     5761 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct
     5821 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca
     5881 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca
     5941 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat
     6001 ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc
     6061 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gataatacca tggtcatgaa
     6121 aacatttaac atttctcaac aagatctaga attagtagaa gtagcgacag agaagattac
     6181 aatgctttat gaggataata aacatcatgt gggagcggca attcgtacga aaacaggaga
     6241 aatcatttcg gcagtacata ttgaagcgta tataggacga gtaactgttt gtgcagaagc
     6301 cattgcgatt ggtagtgcag tttcgaatgg acaaaaggat tttgacacga ttgtagctgt
     6361 tagacaccct tattctgacg aagtagatag aagtattcga gtggtaagtc cttgtggtat
     6421 gtgtagggag ttgatttcag actatgcacc agattgtttt gtgttaatag aaatgaatgg
     6481 caagttagtc aaaactacga ttgaagaact cattccactc aaatataccc gaaattaaaa
     6541 gtcgagatgc atcgatgatc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     6601 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     6661 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     6721 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     6781 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaacctg cagcccaagc
     6841 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6901 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6961 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     7021 cggatcttaa aagttttgtt actttataga agaaattttg agtttttgtt tttttttaat
     7081 aaataaataa acataaataa attgtttgtt gaatttatta ttagtatgta agtgtaaata
     7141 taataaaact taatatctat tcaaattaat aaataaacct cgatatacag accgataaaa
     7201 cacatgcgtc aattttacgc atgattatct ttaacgtacg tcacaatatg attatctttc
     7261 tagggttaat ctagtatcgt agattaatta atccagcttt tgttcccttt agtgagggtt
     7321 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     7381 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     7441 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     7501 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     7561 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     7621 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     7681 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7741 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7801 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7861 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7921 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7981 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     8041 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     8101 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     8161 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     8221 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     8281 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     8341 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     8401 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     8461 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     8521 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     8581 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     8641 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     8701 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8761 ccgggaagct agagtaagta gttcgc
//
