LOCUS       sequence-485633-        8531 bp    DNA     circular SYN 09-SEP-2025
DEFINITION  Expression of human beta-catenin Armadillo repeats (deletion of a.a.
            2-133 and a.a. 665-781) using piggyBac transposition.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8531)
  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 8531)
  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-Armadillo-HA
FEATURES             Location/Qualifiers
     source          1..8531
                     /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     3395..3417
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             5119..5208
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     misc_feature    5303..5852
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5447..5464)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5674..5693
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(6323..6340)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6329..6553
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6600..6616)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6624..6640
                     /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(6648..6654,6655..6672,6673..6678))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6693..6714
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   6948..7010
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7051..7071)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7051..7069)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7090..7106)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7114..7130
                     /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(7138..7144,7145..7162,7163..7168))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7183..7204
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7321..7338)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7492..8080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7572..7591)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8251..8531,1..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 gcgccagggg gggggggggg gggggggggg gggggggggg
     2281 gggggggggg gggggggggg cggggggggg gggggggggc ggggcgaggg gcggggcggg
     2341 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
     2401 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
     2461 gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg
     2521 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
     2581 tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt
     2641 gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt
     2701 gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg
     2761 cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt
     2821 gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg
     2881 ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct
     2941 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg
     3001 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     3061 cctcgggccg gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt
     3121 cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga
     3181 cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct
     3241 agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     3301 cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg
     3361 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     3421 ggctctagcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc accgacctct
     3481 ctccccaggg ggatccaccg ggctagcgcc accatgcatg cagttgtaaa cttgattaac
     3541 tatcaagatg atgcagaact tgccacacgt gcaatccctg aactgacaaa actgctaaat
     3601 gacgaggacc aggtggtggt taataaggct gcagttatgg tccatcagct ttctaaaaag
     3661 gaagcttcca gacacgctat catgcgttct cctcagatgg tgtctgctat tgtacgtacc
     3721 atgcagaata caaatgatgt agaaacagct cgttgtaccg ctgggacctt gcataacctt
     3781 tcccatcatc gtgagggctt actggccatc tttaagtctg gaggcattcc tgccctggtg
     3841 aaaatgcttg gttcaccagt ggattctgtg ttgttttatg ccattacaac tctccacaac
     3901 cttttattac atcaagaagg agctaaaatg gcagtgcgtt tagctggtgg gctgcagaaa
     3961 atggttgcct tgctcaacaa aacaaatgtt aaattcttgg ctattacgac agactgcctt
     4021 caaattttag cttatggcaa ccaagaaagc aagctcatca tactggctag tggtggaccc
     4081 caagctttag taaatataat gaggacctat acttacgaaa aactactgtg gaccacaagc
     4141 agagtgctga aggtgctatc tgtctgctct agtaataagc cggctattgt agaagctggt
     4201 ggaatgcaag ctttaggact tcacctgaca gatccaagtc aacgtcttgt tcagaactgt
     4261 ctttggactc tcaggaatct ttcagatgct gcaactaaac aggaagggat ggaaggtctc
     4321 cttgggactc ttgttcagct tctgggttca gatgatataa atgtggtcac ctgtgcagct
     4381 ggaattcttt ctaacctcac ttgcaataat tataagaaca agatgatggt ctgccaagtg
     4441 ggtggtatag aggctcttgt gcgtactgtc cttcgggctg gtgacaggga agacatcact
     4501 gagcctgcca tctgtgctct tcgtcatctg accagccgac accaagaagc agagatggcc
     4561 cagaatgcag ttcgccttca ctatggacta ccagttgtgg ttaagctctt acacccacca
     4621 tcccactggc ctctgataaa ggctactgtt ggattgattc gaaatcttgc cctttgtccc
     4681 gcaaatcatg cacctttgcg tgagcagggt gccattccac gactagttca gttgcttgtt
     4741 cgtgcacatc aggataccca gcgccgtacg tccatgggtg ggacacagca gcaatttgtg
     4801 gagggggtcc gcatggaaga aatagttgaa ggttgtaccg gagcccttca catcctagct
     4861 cgggatgttc acaaccgaat tgttatcaga ggactaaata ccattccatt gtttgtgcag
     4921 ctgctttatt ctcccattga aaacatccaa agagtagctg caggggtcct ctgtgaactt
     4981 gctcaggaca aggaagctgc agaagctatt gaagctgagg gagccacagc tcctctgaca
     5041 gagttacttc actctaggaa tgaaggtgtg gcgacatatg cagctgctgt tttgttccga
     5101 atgtctgaga ccggtatgta cccatacgat gttccagatt acgctggcta tccctatgac
     5161 gtcccggact atgcaggctc ctatccatat gacgttccag attacgcttg aacgcgttag
     5221 tgcggatcct agtgcgaatt ctagtgcgtc gactctagca acggtttccc tctagcggga
     5281 tcaattccgc cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     5341 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     5401 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     5461 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
     5521 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     5581 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     5641 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     5701 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
     5761 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     5821 gacgtggttt tcctttgaaa aacacgataa taccatggtc atgaaaacat ttaacatttc
     5881 tcaacaagat ctagaattag tagaagtagc gacagagaag attacaatgc tttatgagga
     5941 taataaacat catgtgggag cggcaattcg tacgaaaaca ggagaaatca tttcggcagt
     6001 acatattgaa gcgtatatag gacgagtaac tgtttgtgca gaagccattg cgattggtag
     6061 tgcagtttcg aatggacaaa aggattttga cacgattgta gctgttagac acccttattc
     6121 tgacgaagta gatagaagta ttcgagtggt aagtccttgt ggtatgtgta gggagttgat
     6181 ttcagactat gcaccagatt gttttgtgtt aatagaaatg aatggcaagt tagtcaaaac
     6241 tacgattgaa gaactcattc cactcaaata tacccgaaat taaaagtcga gatgcatcga
     6301 tgatctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     6361 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     6421 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     6481 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     6541 gtgggctcta tggcttctga ggcggaaaga acctgcagcc caagcttggc gtaatcatgg
     6601 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6661 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6721 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat cttaaaagtt
     6781 ttgttacttt atagaagaaa ttttgagttt ttgttttttt ttaataaata aataaacata
     6841 aataaattgt ttgttgaatt tattattagt atgtaagtgt aaatataata aaacttaata
     6901 tctattcaaa ttaataaata aacctcgata tacagaccga taaaacacat gcgtcaattt
     6961 tacgcatgat tatctttaac gtacgtcaca atatgattat ctttctaggg ttaatctagt
     7021 atcgtagatt aattaatcca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     7081 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     7141 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     7201 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     7261 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     7321 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     7381 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     7441 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7501 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7561 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7621 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     7681 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7741 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7801 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7861 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7921 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7981 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     8041 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     8101 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     8161 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     8221 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     8281 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8341 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8401 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8461 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8521 aagtagttcg c
//
