LOCUS       sequence-481884-        6207 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Expression of GST-tagged human beta-catenin Armadillo repeats ∆4-6 
            without IDRs (deletion of a.a. 2-133, 268-394, and 672-781) in E. 
            coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6207)
  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 6207)
  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: pGEX-5X-1-GST-Armadillo-delta4-6
FEATURES             Location/Qualifiers
     source          1..6207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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)."
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             921..932
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2257..2279)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2424..2442)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2510..2614
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2615..2683,2684..3475)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2833..2852)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3646..4234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4135..4154
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4388..4405
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4478..4555
                     /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             4556..5638
                     /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(4575..4594)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    5651..5672
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5687..5692,5693..5710,5711..5717)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5725..5741
                     /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     5749..5765
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5749..5765
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5761..6207,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5781..5798)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5781..5797)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5790..5812)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg atcgaaggtc gtgggatccc cgaattcatg catgcagttg
      961 taaacttgat taactatcaa gatgatgcag aacttgccac acgtgcaatc cctgaactga
     1021 caaaactgct aaatgacgag gaccaggtgg tggttaataa ggctgcagtt atggtccatc
     1081 agctttctaa aaaggaagct tccagacacg ctatcatgcg ttctcctcag atggtgtctg
     1141 ctattgtacg taccatgcag aatacaaatg atgtagaaac agctcgttgt accgctggga
     1201 ccttgcataa cctttcccat catcgtgagg gcttactggc catctttaag tctggaggca
     1261 ttcctgccct ggtgaaaatg cttggttcac cagtggattc tgtgttgttt tatgccatta
     1321 caactctcca caacctttta ttacatcaag aacaggaagg gatggaaggt ctccttggga
     1381 ctcttgttca gcttctgggt tcagatgata taaatgtggt cacctgtgca gctggaattc
     1441 tttctaacct cacttgcaat aattataaga acaagatgat ggtctgccaa gtgggtggta
     1501 tagaggctct tgtgcgtact gtccttcggg ctggtgacag ggaagacatc actgagcctg
     1561 ccatctgtgc tcttcgtcat ctgaccagcc gacaccaaga agcagagatg gcccagaatg
     1621 cagttcgcct tcactatgga ctaccagttg tggttaagct cttacaccca ccatcccact
     1681 ggcctctgat aaaggctact gttggattga ttcgaaatct tgccctttgt cccgcaaatc
     1741 atgcaccttt gcgtgagcag ggtgccattc cacgactagt tcagttgctt gttcgtgcac
     1801 atcaggatac ccagcgccgt acgtccatgg gtgggacaca gcagcaattt gtggaggggg
     1861 tccgcatgga agaaatagtt gaaggttgta ccggagccct tcacatccta gctcgggatg
     1921 ttcacaaccg aattgttatc agaggactaa ataccattcc attgtttgtg cagctgcttt
     1981 attctcccat tgaaaacatc caaagagtag ctgcaggggt cctctgtgaa cttgctcagg
     2041 acaaggaagc tgcagaagct attgaagctg agggagccac agctcctctg acagagttac
     2101 ttcactctag gaatgaaggt gtggcgacat atgcagctgc tgttttgttc cgaatgtctg
     2161 aggacaagcc acaagattac aagtgagtcg actcgagcgg ccgcatcgtg actgactgac
     2221 gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     2281 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     2341 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     2401 gtataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     2461 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2521 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2581 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2641 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2701 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     2761 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     2821 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     2881 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     2941 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3001 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3061 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3121 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     3181 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3241 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3301 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3361 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3421 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3481 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3541 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3601 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3661 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3721 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3781 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3841 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3901 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3961 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4021 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4081 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4141 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4201 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4261 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4321 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4381 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     4441 tacgcatctg tgcggtattt cacaccgcat aaattccgac accatcgaat ggtgcaaaac
     4501 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
     4561 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
     4621 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
     4681 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
     4741 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
     4801 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
     4861 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
     4921 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
     4981 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
     5041 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
     5101 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
     5161 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
     5221 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
     5281 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
     5341 gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta
     5401 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
     5461 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
     5521 actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt
     5581 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     5641 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     5701 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     5761 atgaccatga ttacggattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     5821 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     5881 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     5941 tttgcctggt ttccggcacc agaagcggtg ccggaaagct ggctggagtg cgatcttcct
     6001 gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga tgcgcccatc
     6061 tacaccaacg taacctatcc cattacggtc aatccgccgt ttgttcccac ggagaatccg
     6121 acgggttgtt actcgctcac atttaatgtt gatgaaagct ggctacagga aggccagacg
     6181 cgaattattt ttgatggcgt tggaatt
//
