LOCUS       sequence-485624-        8836 bp    DNA     circular SYN 09-SEP-2025
DEFINITION  Expression of human beta-catenin ∆arm 4-6 (deletion of a.a. 268-394)
            using piggyBac transposition.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8836)
  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 8836)
  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-delta4-6-HA
FEATURES             Location/Qualifiers
     source          1..8836
                     /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..2393
                     /label=chicken beta-actin promoter
     primer_bind     3334..3356
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      3447..3456
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3498..3533
                     /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             5424..5513
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     misc_feature    5608..6157
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5752..5769)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5979..5998
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(6628..6645)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6634..6858
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6905..6921)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6929..6945
                     /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(6953..6959,6960..6977,6978..6983))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6998..7019
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   7253..7315
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7356..7376)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7356..7374)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7395..7411)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7419..7435
                     /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(7443..7449,7450..7467,7468..7473))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7488..7509
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7626..7643)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7797..8385)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7877..7896)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8556..8836,1..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 gggggggcgc gcgccaggcg ggggggggcg gggcgagggg cggggcgggg
     2281 cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta
     2341 tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg
     2401 ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc
     2461 tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct
     2521 gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg
     2581 aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg
     2641 tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc
     2701 gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg
     2761 ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg
     2821 gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg cacccccctc
     2881 cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg
     2941 ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc
     3001 ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc
     3061 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac
     3121 ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta
     3181 gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc
     3241 gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg
     3301 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     3361 gctctagccg gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc
     3421 tccccagggg gatccaccgg gctagcgcca ccatggctac tcaagctgat ttgatggagt
     3481 tggacatggc catggaacca gacagaaaag cggctgttag tcactggcag caacagtctt
     3541 acctggactc tggaatccat tctggtgcca ctaccacagc tccttctctg agtggtaaag
     3601 gcaatcctga ggaagaggat gtggatacct cccaagtcct gtatgagtgg gaacagggat
     3661 tttctcagtc cttcactcaa gaacaagtag ctgatattga tggacagtat gcaatgactc
     3721 gagctcagag ggtacgagct gctatgttcc ctgagacatt agatgagggc atgcagatcc
     3781 catctacaca gtttgatgct gctcatccca ctaatgtcca gcgtttggct gaaccatcac
     3841 agatgctgaa acatgcagtt gtaaacttga ttaactatca agatgatgca gaacttgcca
     3901 cacgtgcaat ccctgaactg acaaaactgc taaatgacga ggaccaggtg gtggttaata
     3961 aggctgcagt tatggtccat cagctttcta aaaaggaagc ttccagacac gctatcatgc
     4021 gttctcctca gatggtgtct gctattgtac gtaccatgca gaatacaaat gatgtagaaa
     4081 cagctcgttg taccgctggg accttgcata acctttccca tcatcgtgag ggcttactgg
     4141 ccatctttaa gtctggaggc attcctgccc tggtgaaaat gcttggttca ccagtggatt
     4201 ctgtgttgtt ttatgccatt acaactctcc acaacctttt attacatcaa gaacaggaag
     4261 ggatggaagg tctccttggg actcttgttc agcttctggg ttcagatgat ataaatgtgg
     4321 tcacctgtgc agctggaatt ctttctaacc tcacttgcaa taattataag aacaagatga
     4381 tggtctgcca agtgggtggt atagaggctc ttgtgcgtac tgtccttcgg gctggtgaca
     4441 gggaagacat cactgagcct gccatctgtg ctcttcgtca tctgaccagc cgacaccaag
     4501 aagcagagat ggcccagaat gcagttcgcc ttcactatgg actaccagtt gtggttaagc
     4561 tcttacaccc accatcccac tggcctctga taaaggctac tgttggattg attcgaaatc
     4621 ttgccctttg tcccgcaaat catgcacctt tgcgtgagca gggtgccatt ccacgactag
     4681 ttcagttgct tgttcgtgca catcaggata cccagcgccg tacgtccatg ggtgggacac
     4741 agcagcaatt tgtggagggg gtccgcatgg aagaaatagt tgaaggttgt accggagccc
     4801 ttcacatcct agctcgggat gttcacaacc gaattgttat cagaggacta aataccattc
     4861 cattgtttgt gcagctgctt tattctccca ttgaaaacat ccaaagagta gctgcagggg
     4921 tcctctgtga acttgctcag gacaaggaag ctgcagaagc tattgaagct gagggagcca
     4981 cagctcctct gacagagtta cttcactcta ggaatgaagg tgtggcgaca tatgcagctg
     5041 ctgttttgtt ccgaatgtct gaggacaagc cacaagatta caagaaacgg ctttcagttg
     5101 agctgaccag ctctctcttc agaacagagc caatggcttg gaatgagact gctgatcttg
     5161 gacttgatat tggtgcccag ggagaacccc ttggatatcg ccaggatgat cctagctatc
     5221 gttcttttca ctctggtgga tatggccagg atgccttggg tatggacccc atgatggaac
     5281 atgagatggg tggccaccac cctggtgctg actatccagt tgatgggctg ccagatctgg
     5341 ggcatgccca ggacctcatg gatgggctgc ctccaggtga cagcaatcag ctggcctggt
     5401 ttgatactga cctgaccggt atgtacccat acgatgttcc agattacgct ggctatccct
     5461 atgacgtccc ggactatgca ggctcctatc catatgacgt tccagattac gcttgaacgc
     5521 gttagtgcgg atcctagtgc gaattctagt gcgtcgactc tagcaacggt ttccctctag
     5581 cgggatcaat tccgcccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc
     5641 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
     5701 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca
     5761 aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
     5821 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
     5881 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
     5941 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
     6001 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
     6061 ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
     6121 acggggacgt ggttttcctt tgaaaaacac gataatacca tggtcatgaa aacatttaac
     6181 atttctcaac aagatctaga attagtagaa gtagcgacag agaagattac aatgctttat
     6241 gaggataata aacatcatgt gggagcggca attcgtacga aaacaggaga aatcatttcg
     6301 gcagtacata ttgaagcgta tataggacga gtaactgttt gtgcagaagc cattgcgatt
     6361 ggtagtgcag tttcgaatgg acaaaaggat tttgacacga ttgtagctgt tagacaccct
     6421 tattctgacg aagtagatag aagtattcga gtggtaagtc cttgtggtat gtgtagggag
     6481 ttgatttcag actatgcacc agattgtttt gtgttaatag aaatgaatgg caagttagtc
     6541 aaaactacga ttgaagaact cattccactc aaatataccc gaaattaaaa gtcgagatgc
     6601 atcgatgatc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     6661 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     6721 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     6781 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     6841 atgcggtggg ctctatggct tctgaggcgg aaagaacctg cagcccaagc ttggcgtaat
     6901 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     6961 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     7021 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatcttaa
     7081 aagttttgtt actttataga agaaattttg agtttttgtt tttttttaat aaataaataa
     7141 acataaataa attgtttgtt gaatttatta ttagtatgta agtgtaaata taataaaact
     7201 taatatctat tcaaattaat aaataaacct cgatatacag accgataaaa cacatgcgtc
     7261 aattttacgc atgattatct ttaacgtacg tcacaatatg attatctttc tagggttaat
     7321 ctagtatcgt agattaatta atccagcttt tgttcccttt agtgagggtt aattgcgcgc
     7381 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     7441 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     7501 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     7561 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     7621 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     7681 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     7741 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7801 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7861 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     7921 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7981 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     8041 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     8101 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     8161 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     8221 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     8281 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     8341 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     8401 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     8461 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     8521 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     8581 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     8641 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     8701 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     8761 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     8821 agagtaagta gttcgc
//
