LOCUS       sequence-485625-        8875 bp    DNA     circular SYN 09-SEP-2025
DEFINITION  Expression of core binding region (CBR) of human ARID1A using 
            piggyBac transposition. ∆IDR1, ∆ARID, and ∆IDR2 (deletion of a.a 
            2-1610).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8875)
  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 8875)
  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-ARID1A-CBR-FLAG
FEATURES             Location/Qualifiers
     source          1..8875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     127..149
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     141..158
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     142..158
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     168..187
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        168..186
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   216..250
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        571..950
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        952..1227
                     /label=chicken beta-actin promoter
     primer_bind     2168..2190
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             4321..4386
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    4481..5030
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4625..4642)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4852..4871
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(5501..5518)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5507..5731
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5778..5794)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5802..5818
                     /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(5826..5832,5833..5850,5851..5856))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5871..5892
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   6126..6188
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6229..6249)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6229..6247)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6268..6284)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6292..6308
                     /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(6316..6322,6323..6340,6341..6346))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6361..6382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6499..6516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6670..7258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6750..6769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7429..8220,8221..8289))
                     /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"
     primer_bind     8052..8071
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8290..8394)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8420..8875)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8557..8578)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8769..8788
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
      121 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac
      181 tatagggcga attggggcgc gccattctag attaacccta gaaagatagt ctgcgtaaaa
      241 ttgacgcatg cattcttgaa atattgctct ctctttctaa atagcgcgaa tccgtcgctg
      301 tgcatttagg acatctcagt cgccgcttgg agctcccgtg aggcgtgctt gtcaatgcgg
      361 taagtgtcac tgattttgaa ctataacgac cgcgtgagtc aaaatgacgc atgattatct
      421 tttacgtgac ttttaagatt taactcatac gataattata ttgttatttc atgttctact
      481 tacgtgataa cttattatat atatattttc ttgttataga tatcaactag aatgctagac
      541 tcgagaagct tgatcggcca cgcggatctc gacattgatt attgactagt tattaatagt
      601 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      661 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      721 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      781 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      841 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      901 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga
      961 gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt
     1021 tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg gcgcgggcgg
     1081 gggggggggg gggggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc
     1141 aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc
     1201 tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc
     1261 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
     1321 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg
     1381 cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc
     1441 ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc
     1501 tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc
     1561 agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag
     1621 gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg
     1681 tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct
     1741 tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg
     1801 cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag
     1861 gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc
     1921 cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag
     1981 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc
     2041 gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt
     2101 ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc
     2161 agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gccggagcgc acgtcggcag
     2221 tcggctccct cgttgaccga atcaccgacc tctctcccca gggggatcca ccgggctagc
     2281 gccaccatga tgaaaatgca gaaggcaggt cccccagtac ctgcctcgca catagcacct
     2341 gcccctgtgc agccccccat gattcggcgg gatatcacct tcccacctgg ctctgttgaa
     2401 gccacacagc ctgtgttgaa gcagaggagg cggctcacaa tgaaagacat tggaaccccg
     2461 gaggcatggc gggtaatgat gtccctcaag tctggtctcc tggcagagag cacatgggca
     2521 ttagatacca tcaacatcct gctgtatgat gacaacagca tcatgacctt caacctcagt
     2581 cagctcccag ggttgctaga gctccttgta gaatatttcc gacgatgcct gattgagatc
     2641 tttggcattt taaaggagta tgaggtgggt gacccaggac agagaacgct actggatcct
     2701 gggaggttca gcaaggtgtc tagtccagct cccatggagg gtggggaaga agaagaagaa
     2761 cttctaggtc ctaaactaga agaggaagaa gaagaggaag tagttgaaaa tgatgaggag
     2821 atagcctttt caggcaagga caagccagct tcagagaata gtgaggagaa gctgatcagt
     2881 aagtttgaca agcttccagt aaagatcgta cagaagaatg atccatttgt ggtggactgc
     2941 tcagataagc ttgggcgtgt gcaggagttt gacagtggcc tgctgcactg gcggattggt
     3001 gggggggaca ccactgagca tatccagacc cacttcgaga gcaagacaga gctgctgcct
     3061 tcccggcctc acgcaccctg cccaccagcc cctcggaagc atgtgacaac agcagagggt
     3121 acaccaggga caacagacca ggaggggccc ccacctgatg gacctccaga aaaacggatc
     3181 acagccacta tggatgacat gttgtctact cggtctagca ccttgaccga ggatggagct
     3241 aagagttcag aggccatcaa ggagagcagc aagtttccat ttggcattag cccagcacag
     3301 agccaccgga acatcaagat cctagaggac gaaccccaca gtaaggatga gaccccactg
     3361 tgtacccttc tggactggca ggattctctt gccaagcgct gcgtctgtgt gtccaatacc
     3421 attcgaagcc tgtcatttgt gccaggcaat gactttgaga tgtccaaaca cccagggctg
     3481 ctgctcatcc tgggcaagct gatcctgctg caccacaagc acccagaacg gaagcaggca
     3541 ccactaactt atgaaaagga ggaggaacag gaccaagggg tgagctgcaa caaagtggag
     3601 tggtggtggg actgcttgga gatgctccgg gaaaacacct tggttacact cgccaacatc
     3661 tcggggcagt tggacctatc tccatacccc gagagcattt gcctgcctgt cctggacgga
     3721 ctcctacact gggcagtttg cccttcagct gaagcccagg accccttttc caccctgggc
     3781 cccaatgccg tcctttcccc gcagagactg gtcttggaaa ccctcagcaa actcagcatc
     3841 caggacaaca atgtggacct gattctggcc acacccccct tcagccgcct ggagaagttg
     3901 tatagcacta tggtgcgctt cctcagtgac cgaaagaacc cggtgtgccg ggagatggct
     3961 gtggtactgc tggccaacct ggctcagggg gacagcctgg cagctcgtgc cattgcagtg
     4021 cagaagggca gtatcggcaa cctcctgggc ttcctagagg acagccttgc cgccacacag
     4081 ttccagcaga gccaggccag cctcctccac atgcagaacc caccctttga gccaactagt
     4141 gtggacatga tgcggcgggc tgcccgcgcg ctgcttgcct tggccaaggt ggacgagaac
     4201 cactcagagt ttactctgta cgaatcacgg ctgttggaca tctcggtatc accgctgatg
     4261 aatagcctcg tgtcacaagt catttgtgat gtactgtttt tgattggcca gtccaccggt
     4321 gactacaaag accatgacgg tgattataaa gatcatgata tcgattacaa ggatgacgat
     4381 gacaagtgaa cgcgttagtg cggatcctag tgcgaattct agtgcgtcga ctctagcaac
     4441 ggtttccctc tagcgggatc aattccgccc ccccccccta acgttactgg ccgaagccgc
     4501 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     4561 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     4621 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg
     4681 gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     4741 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     4801 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     4861 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     4921 gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
     4981 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgataata ccatggtcat
     5041 gaaaacattt aacatttctc aacaagatct agaattagta gaagtagcga cagagaagat
     5101 tacaatgctt tatgaggata ataaacatca tgtgggagcg gcaattcgta cgaaaacagg
     5161 agaaatcatt tcggcagtac atattgaagc gtatatagga cgagtaactg tttgtgcaga
     5221 agccattgcg attggtagtg cagtttcgaa tggacaaaag gattttgaca cgattgtagc
     5281 tgttagacac ccttattctg acgaagtaga tagaagtatt cgagtggtaa gtccttgtgg
     5341 tatgtgtagg gagttgattt cagactatgc accagattgt tttgtgttaa tagaaatgaa
     5401 tggcaagtta gtcaaaacta cgattgaaga actcattcca ctcaaatata cccgaaatta
     5461 aaagtcgaga tgcatcgatg atctagagct cgctgatcag cctcgactgt gccttctagt
     5521 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     5581 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     5641 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     5701 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac ctgcagccca
     5761 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5821 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5881 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5941 cagcggatct taaaagtttt gttactttat agaagaaatt ttgagttttt gttttttttt
     6001 aataaataaa taaacataaa taaattgttt gttgaattta ttattagtat gtaagtgtaa
     6061 atataataaa acttaatatc tattcaaatt aataaataaa cctcgatata cagaccgata
     6121 aaacacatgc gtcaatttta cgcatgatta tctttaacgt acgtcacaat atgattatct
     6181 ttctagggtt aatctagtat cgtagattaa ttaatccagc ttttgttccc tttagtgagg
     6241 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6301 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6361 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6421 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6481 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6541 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6601 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6661 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6721 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6781 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6841 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6901 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6961 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7021 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7081 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     7141 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7201 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7261 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7321 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7381 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7441 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7501 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7561 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7621 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7681 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7741 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7801 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7861 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7921 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7981 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8041 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8101 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8161 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8221 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8281 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8341 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8401 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     8461 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     8521 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     8581 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     8641 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     8701 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     8761 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     8821 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgt
//
