LOCUS       sequence-481888-        6588 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Expression of GST-tagged human beta-catenin Armadillo repeats 
            without IDRs (deletion of a.a. 2-133 and 672-781) in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6588)
  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 6588)
  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
FEATURES             Location/Qualifiers
     source          1..6588
                     /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(2638..2660)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2805..2823)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2891..2995
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2996..3064,3065..3856)
                     /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(3214..3233)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4027..4615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4516..4535
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4769..4786
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4859..4936
                     /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             4937..6019
                     /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(4956..4975)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    6032..6053
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6068..6073,6074..6091,6092..6098)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6106..6122
                     /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     6130..6146
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6130..6146
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6142..6588,1..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(6162..6179)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6162..6178)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6171..6193)
                     /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 aaggagctaa aatggcagtg cgtttagctg
     1381 gtgggctgca gaaaatggtt gccttgctca acaaaacaaa tgttaaattc ttggctatta
     1441 cgacagactg ccttcaaatt ttagcttatg gcaaccaaga aagcaagctc atcatactgg
     1501 ctagtggtgg accccaagct ttagtaaata taatgaggac ctatacttac gaaaaactac
     1561 tgtggaccac aagcagagtg ctgaaggtgc tatctgtctg ctctagtaat aagccggcta
     1621 ttgtagaagc tggtggaatg caagctttag gacttcacct gacagatcca agtcaacgtc
     1681 ttgttcagaa ctgtctttgg actctcagga atctttcaga tgctgcaact aaacaggaag
     1741 ggatggaagg tctccttggg actcttgttc agcttctggg ttcagatgat ataaatgtgg
     1801 tcacctgtgc agctggaatt ctttctaacc tcacttgcaa taattataag aacaagatga
     1861 tggtctgcca agtgggtggt atagaggctc ttgtgcgtac tgtccttcgg gctggtgaca
     1921 gggaagacat cactgagcct gccatctgtg ctcttcgtca tctgaccagc cgacaccaag
     1981 aagcagagat ggcccagaat gcagttcgcc ttcactatgg actaccagtt gtggttaagc
     2041 tcttacaccc accatcccac tggcctctga taaaggctac tgttggattg attcgaaatc
     2101 ttgccctttg tcccgcaaat catgcacctt tgcgtgagca gggtgccatt ccacgactag
     2161 ttcagttgct tgttcgtgca catcaggata cccagcgccg tacgtccatg ggtgggacac
     2221 agcagcaatt tgtggagggg gtccgcatgg aagaaatagt tgaaggttgt accggagccc
     2281 ttcacatcct agctcgggat gttcacaacc gaattgttat cagaggacta aataccattc
     2341 cattgtttgt gcagctgctt tattctccca ttgaaaacat ccaaagagta gctgcagggg
     2401 tcctctgtga acttgctcag gacaaggaag ctgcagaagc tattgaagct gagggagcca
     2461 cagctcctct gacagagtta cttcactcta ggaatgaagg tgtggcgaca tatgcagctg
     2521 ctgttttgtt ccgaatgtct gaggacaagc cacaagatta caagtgagtc gactcgagcg
     2581 gccgcatcgt gactgactga cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     2641 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     2701 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     2761 gtcacgtagc gatagcggag tgtataattc ttgaagacga aagggcctcg tgatacgcct
     2821 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     2881 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2941 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3001 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3061 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3121 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3181 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     3241 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3301 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3361 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3421 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3481 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     3541 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3601 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3661 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3721 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3781 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3841 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3901 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3961 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4021 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4081 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4141 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     4201 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4261 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4321 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4381 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4441 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4501 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4561 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4621 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4681 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4741 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4801 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca taaattccga
     4861 caccatcgaa tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
     4921 attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
     4981 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
     5041 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca
     5101 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
     5161 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
     5221 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
     5281 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
     5341 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
     5401 catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt
     5461 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
     5521 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga
     5581 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga
     5641 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg
     5701 cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga
     5761 taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct
     5821 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg
     5881 caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca
     5941 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     6001 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     6061 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     6121 aatttcacac aggaaacagc tatgaccatg attacggatt cactggccgt cgttttacaa
     6181 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     6241 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     6301 agcctgaatg gcgaatggcg ctttgcctgg tttccggcac cagaagcggt gccggaaagc
     6361 tggctggagt gcgatcttcc tgaggccgat actgtcgtcg tcccctcaaa ctggcagatg
     6421 cacggttacg atgcgcccat ctacaccaac gtaacctatc ccattacggt caatccgccg
     6481 tttgttccca cggagaatcc gacgggttgt tactcgctca catttaatgt tgatgaaagc
     6541 tggctacagg aaggccagac gcgaattatt tttgatggcg ttggaatt
//
