LOCUS       sequence-481885-        7974 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Expression of MBP-tagged human ARID1A IDR2 (a.a. 1170-1610) in E. 
            coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7974)
  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 7974)
  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: pMAL-c4X-MBP-ARID1A-IDR2
FEATURES             Location/Qualifiers
     source          1..7974
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..80
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             81..1163
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(100..119)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1406..1411,1412..1427,1428..1434)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1442..1458
                     /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)."
     primer_bind     1447..1469
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1528..2628
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     2602..2625
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2677..2688
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4065..4082)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4065..4081)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4074..4096)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4260..4277)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4430..4516
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4608..4635
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4654..4745
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4746..4814,4815..5606)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4964..4983)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5648..6161)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5843..5864)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6055..6074
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      6272..6860
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6761..6780
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7014..7031
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7046..7188
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7114..7133)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7274..7296
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7290..7481)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
       61 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
      121 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
      181 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
      241 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
      301 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
      361 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
      421 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
      481 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
      541 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
      601 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
      661 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
      721 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
      781 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
      841 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
      901 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
      961 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     1021 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     1081 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     1141 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     1201 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg
     1261 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     1321 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     1381 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
     1441 attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga
     1501 gcacttcacc aacaaggacc atagcatatg aaaatcgaag aaggtaaact ggtaatctgg
     1561 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat
     1621 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt
     1681 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac
     1741 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat
     1801 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt
     1861 gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa
     1921 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac
     1981 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag
     2041 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg
     2101 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac
     2161 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg
     2221 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc
     2281 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt
     2341 ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg
     2401 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag
     2461 ttggtgaaag atccgcggat tgccgccact atggaaaacg cccagaaagg tgaaatcatg
     2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc
     2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg
     2641 aacaacaaca acaataacaa taacaacaac ctcgggatcg agggaaggat ttcagaattc
     2701 ggatccaccg gtatgcacag tcagatcccc ccattgccag gcatgagcag gagcaattca
     2761 gttgggatcc aggatgcctt taatgatgga agtgactcca cattccagaa gcggaattcc
     2821 atgactccaa accctgggta tcagcccagt atgaatacct ctgacatgat ggggcgcatg
     2881 tcctatgagc caaataagga tccttatggc agcatgagga aagctccagg gagtgatccc
     2941 ttcatgtcct cagggcaggg ccccaacggc gggatgggtg acccctacag tcgtgctgcc
     3001 ggccctgggc taggaaatgt ggcgatggga ccacgacagc actatcccta tggaggtcct
     3061 tatgacagag tgaggacgga gcctggaata gggcctgagg gaaacatgag cactggggcc
     3121 ccacagccga atctcatgcc ttccaaccca gactcgggga tgtattctcc tagccgctac
     3181 cccccgcagc agcagcagca gcagcagcaa cgacatgatt cctatggcaa tcagttctcc
     3241 acccaaggca ccccttctgg cagccccttc cccagccagc agactacaat gtatcaacag
     3301 caacagcaga attacaagcg gccaatggat ggcacatatg gccctcctgc caagcggcac
     3361 gaaggggaga tgtacagcgt gccatacagc actgggcagg ggcagcctca gcagcagcag
     3421 ttgcccccag cccagcccca gcctgccagc cagcaacaag ctgcccagcc ttcccctcag
     3481 caagatgtat acaaccagta tggcaatgcc tatcctgcca ctgccacagc tgctactgag
     3541 cgccgaccag caggcggccc ccagaaccaa tttccattcc agtttggccg agaccgtgtc
     3601 tctgcacccc ctggcaccaa tgcccagcaa aacatgccac cacaaatgat gggcggcccc
     3661 atacaggcat cagctgaggt tgctcagcaa ggcaccatgt ggcaggggcg taatgacatg
     3721 acctataatt atgccaacag gcagagcacg ggctctgccc cccagggccc cgcctatcat
     3781 ggcgtgaacc gaacagatga aatgctgcac acagatcaga gggccaacca cgaaggctcg
     3841 tggccttccc atggcacacg ccagccccca tatggtccct ctgcccctgt gccccccatg
     3901 acaaggcccc ctccatctaa ctaccagccc ccaccaagca tgcagaatca cattcctcag
     3961 gtatccagcc ctgctcccct gccccggcca atggagaacc gcacctctcc tagcaagtct
     4021 ccattcctgc actctgggtg agtcgacctg caggcaagct tggcactggc cgtcgtttta
     4081 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     4141 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     4201 cgcagcctga atggcgaatg gcagcttggc tgttttggcg gatgagataa gattttcagc
     4261 ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc
     4321 agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc
     4381 gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg
     4441 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     4501 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg
     4561 gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct
     4621 gacggatggc ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa
     4681 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4741 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4801 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4861 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4921 gccccgaaga acgtttccca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4981 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5041 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5101 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5161 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5221 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5281 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5341 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5401 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5461 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5521 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5581 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5641 ttgatttacc ccggttgata atcagaaaag ccccaaaaac aggaagattg tataagcaaa
     5701 tatttaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     5761 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     5821 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     5881 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     5941 acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     6001 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     6061 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     6121 caccacaccc gccgcgctta atgcgccgct acagggcgcg taaaaggatc taggtgaaga
     6181 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6241 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6301 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6361 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     6421 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6481 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6541 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6601 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6661 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6721 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6781 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6841 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6901 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6961 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     7021 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     7081 gtatttcaca ccgcatatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     7141 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     7201 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     7261 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     7321 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgca gcgattcaca gatgtctgcc
     7381 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     7441 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     7501 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     7561 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     7621 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     7681 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     7741 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     7801 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     7861 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     7921 gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga aatt
//
