LOCUS       sequence-481881-        8839 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Expression of human beta-catenin ∆arm 7-9 (deletion of a.a. 395-520)
            using piggyBac transposition.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8839)
  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 8839)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 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-delta7-9-HA
FEATURES             Location/Qualifiers
     source          1..8839
                     /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             5427..5516
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     misc_feature    5611..6160
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5755..5772)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5982..6001
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(6631..6648)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6637..6861
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6908..6924)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6932..6948
                     /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(6956..6962,6963..6980,6981..6986))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7001..7022
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   7256..7318
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7359..7379)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7359..7377)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7398..7414)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7422..7438
                     /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(7446..7452,7453..7470,7471..7476))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7491..7512
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7629..7646)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7800..8388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7880..7899)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8559..8839,1..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 gggcggggcg 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 gaaggagcta
     4261 aaatggcagt gcgtttagct ggtgggctgc agaaaatggt tgccttgctc aacaaaacaa
     4321 atgttaaatt cttggctatt acgacagact gccttcaaat tttagcttat ggcaaccaag
     4381 aaagcaagct catcatactg gctagtggtg gaccccaagc tttagtaaat ataatgagga
     4441 cctatactta cgaaaaacta ctgtggacca caagcagagt gctgaaggtg ctatctgtct
     4501 gctctagtaa taagccggct attgtagaag ctggtggaat gcaagcttta ggacttcacc
     4561 tgacagatcc aagtcaacgt cttgttcaga actgtctttg gactctcagg aatctttcag
     4621 atgctgcaac taaacccgca aatcatgcac ctttgcgtga gcagggtgcc attccacgac
     4681 tagttcagtt gcttgttcgt gcacatcagg atacccagcg ccgtacgtcc atgggtggga
     4741 cacagcagca atttgtggag ggggtccgca tggaagaaat agttgaaggt tgtaccggag
     4801 cccttcacat cctagctcgg gatgttcaca accgaattgt tatcagagga ctaaatacca
     4861 ttccattgtt tgtgcagctg ctttattctc ccattgaaaa catccaaaga gtagctgcag
     4921 gggtcctctg tgaacttgct caggacaagg aagctgcaga agctattgaa gctgagggag
     4981 ccacagctcc tctgacagag ttacttcact ctaggaatga aggtgtggcg acatatgcag
     5041 ctgctgtttt gttccgaatg tctgaggaca agccacaaga ttacaagaaa cggctttcag
     5101 ttgagctgac cagctctctc ttcagaacag agccaatggc ttggaatgag actgctgatc
     5161 ttggacttga tattggtgcc cagggagaac cccttggata tcgccaggat gatcctagct
     5221 atcgttcttt tcactctggt ggatatggcc aggatgcctt gggtatggac cccatgatgg
     5281 aacatgagat gggtggccac caccctggtg ctgactatcc agttgatggg ctgccagatc
     5341 tggggcatgc ccaggacctc atggatgggc tgcctccagg tgacagcaat cagctggcct
     5401 ggtttgatac tgacctgacc ggtatgtacc catacgatgt tccagattac gctggctatc
     5461 cctatgacgt cccggactat gcaggctcct atccatatga cgttccagat tacgcttgaa
     5521 cgcgttagtg cggatcctag tgcgaattct agtgcgtcga ctctagcaac ggtttccctc
     5581 tagcgggatc aattccgccc ccccccccta acgttactgg ccgaagccgc ttggaataag
     5641 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     5701 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     5761 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     5821 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     5881 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     5941 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     6001 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     6061 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     6121 accacgggga cgtggttttc ctttgaaaaa cacgataata ccatggtcat gaaaacattt
     6181 aacatttctc aacaagatct agaattagta gaagtagcga cagagaagat tacaatgctt
     6241 tatgaggata ataaacatca tgtgggagcg gcaattcgta cgaaaacagg agaaatcatt
     6301 tcggcagtac atattgaagc gtatatagga cgagtaactg tttgtgcaga agccattgcg
     6361 attggtagtg cagtttcgaa tggacaaaag gattttgaca cgattgtagc tgttagacac
     6421 ccttattctg acgaagtaga tagaagtatt cgagtggtaa gtccttgtgg tatgtgtagg
     6481 gagttgattt cagactatgc accagattgt tttgtgttaa tagaaatgaa tggcaagtta
     6541 gtcaaaacta cgattgaaga actcattcca ctcaaatata cccgaaatta aaagtcgaga
     6601 tgcatcgatg atctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     6661 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     6721 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     6781 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     6841 gggatgcggt gggctctatg gcttctgagg cggaaagaac ctgcagccca agcttggcgt
     6901 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6961 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     7021 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatct
     7081 taaaagtttt gttactttat agaagaaatt ttgagttttt gttttttttt aataaataaa
     7141 taaacataaa taaattgttt gttgaattta ttattagtat gtaagtgtaa atataataaa
     7201 acttaatatc tattcaaatt aataaataaa cctcgatata cagaccgata aaacacatgc
     7261 gtcaatttta cgcatgatta tctttaacgt acgtcacaat atgattatct ttctagggtt
     7321 aatctagtat cgtagattaa ttaatccagc ttttgttccc tttagtgagg gttaattgcg
     7381 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     7441 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     7501 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     7561 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     7621 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     7681 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     7741 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     7801 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     7861 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7921 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7981 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     8041 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     8101 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     8161 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     8221 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     8281 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     8341 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     8401 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     8461 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     8521 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     8581 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     8641 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     8701 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     8761 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     8821 gctagagtaa gtagttcgc
//
