LOCUS       sequence-482262-        6013 bp    DNA     circular SYN 06-AUG-2025
DEFINITION  Expesses GST-TRIP12 for protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6013)
  AUTHORS   Kassel S, Yuan K, Bunnag N, Neitzel LR, Lu W, Schwarzkopf A, Maines 
            B, Loberg MA, Xu G, Adams A, McCray AD, Cho A, Rockouski M, Orton G,
            Goldsmith L, Aronno MMA, Spencer ZT, Khan OM, Ye F, Williams C, 
            Lebensohn AM, Rohatgi R, Wang X, Weiss VL, Hong CC, Kettenbach AN, 
            Robbins DJ, Ahmed Y, Lee E
  TITLE     The TRIP12 E3 ligase induces SWI/SNF component BRG1-beta-catenin 
            interaction to promote Wnt signaling.
  JOURNAL   Nat Commun. 2025 Jun 5;16(1):5248. doi: 10.1038/s41467-025-60535-5.
  PUBMED    40473626
REFERENCE   2  (bases 1 to 6013)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2025-06-5"; volume: "16(1):5248. doi"; pages: " 
            10.1038/s41467-025-60535-5"
COMMENT     Sequence Label: pGEX-TRIP12-HECT
FEATURES             Location/Qualifiers
     source          1..6013
                     /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             918..935
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2063..2085)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2230..2248)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2316..2420
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2421..2489,2490..3281)
                     /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(2639..2658)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3452..4040
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3941..3960
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4194..4211
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4284..4361
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4362..5444
                     /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(4381..4400)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5493..5498,5499..5516,5517..5523)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5531..5547
                     /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     5555..5571
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5555..5571
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5567..6013,1..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(5587..5604)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5587..5603)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5596..5618)
                     /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 gttccgcgtg gatccccgga attcatgcag cgtgcagatc
      961 tgggcctgtg gcgcggcgaa gaagttaccc tgagcaatcc gaaaggtagt caggaaggta
     1021 ccaaatatat tcagaatctg cagggcctgt ttgcactgcc gtttggccgc accgcaaaac
     1081 cggcccatat tgccaaagtg aaaatgaaat ttcgctttct gggtaaactg atggccaaag
     1141 caattatgga ttttcgtctg gtggatctgc cgctgggtct gccgttttat aaatggatgc
     1201 tgcgtcagga aaccagtctg accagtcatg atctgtttga tattgatccg gttgtggcac
     1261 gtagtgtgta tcatctggaa gatattgtgc gccagaaaaa acgtctggaa caggataaaa
     1321 gtcagaccaa agaaagtctg cagtatgcac tggaaaccct gaccatgaat ggctgtagtg
     1381 ttgaagatct gggcttagat tttaccctgc cgggctttcc gaatattgaa ctgaaaaaag
     1441 gtggcaaaga tattccggtt accattcata atctggaaga atatctgcgt ctggttattt
     1501 tctgggcact gaatgaaggt gtgagccgcc agtttgatag ttttcgcgat ggctttgaaa
     1561 gtgtgtttcc gctgagtcat ctgcagtatt tttatccgga agaactggat cagctgctgt
     1621 gcggtagtaa agccgatacc tgggatgcca aaaccctgat ggaatgttgt cgcccggatc
     1681 atggctatac ccatgatagc cgcgcagtga aatttctgtt tgaaattctg agtagtttcg
     1741 ataatgagca gcagcgtctg tttctgcagt ttgttaccgg cagtccgcgt ctgccggtgg
     1801 gcggttttcg cagcctgaat ccgccgctga ccattgtgcg caaaaccttt gaaagtaccg
     1861 aaaatccgga tgattttctg ccgagtgtga tgacctgcgt taattatctg aaactgccgg
     1921 attatagtag tattgaaatt atgcgtgaga agctgctgat tgccgcccgc gaaggtcagc
     1981 agagttttca tctgagcctc gagcggccgc atcgtgactg actgacgatc tgcctcgcgc
     2041 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     2101 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2161 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat aattcttgaa
     2221 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     2281 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2341 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2401 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     2461 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2521 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2581 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     2641 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     2701 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     2761 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     2821 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2881 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2941 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     3001 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3061 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3121 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3181 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3241 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3301 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3361 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3421 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3481 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3541 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     3601 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3661 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3721 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3781 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3841 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3901 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3961 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4021 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4081 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4141 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     4201 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     4261 gtatttcaca ccgcataaat tccgacacca tcgaatggcg caaaaccttt cgcggtatgg
     4321 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
     4381 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     4441 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
     4501 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
     4561 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
     4621 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
     4681 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
     4741 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat
     4801 ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat gaagacggta
     4861 cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg
     4921 gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc
     4981 gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc
     5041 aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg
     5101 atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg
     5161 atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa
     5221 ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac
     5281 tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa
     5341 aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     5401 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     5461 gtaagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     5521 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     5581 ggattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     5641 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     5701 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc
     5761 ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt
     5821 cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtgac
     5881 ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc
     5941 gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga
     6001 tggcgttgga att
//
