LOCUS       sequence-209451-        6634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant GST tagged protein production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6634)
  AUTHORS   Coelho MB, Attig J, Bellora N, Konig J, Hallegger M, Kayikci M, 
            Eyras E, Ule J, Smith CW
  TITLE     Nuclear matrix protein Matrin3 regulates alternative splicing and 
            forms overlapping regulatory networks with PTB.
  JOURNAL   EMBO J. 2015 Mar 4;34(5):653-68. doi: 10.15252/embj.201489852. Epub 
            2015 Jan 19.
  PUBMED    25599992
REFERENCE   2  (bases 1 to 6634)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.15252/embj.201489852"; journalName: "EMBO J"; date: "2015-03-4- 
            4"; volume: "34"; issue: "5"; pages: "653-68"
COMMENT     Sequence Label: GST PTBP1 isoform 4
FEATURES             Location/Qualifiers
     source          1..6634
                     /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(2684..2706)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2851..2869)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2937..3041
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3042..3110,3111..3902)
                     /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(3260..3279)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4073..4661
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4562..4581
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4815..4832
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4905..4982
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4983..6065
                     /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(5002..5021)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6114..6119,6120..6137,6138..6144)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6152..6168
                     /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     6176..6192
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6176..6192
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6188..6634,1..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(6208..6225)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6208..6224)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6217..6239)
                     /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 attcatggac ggcattgtcc
      961 cagatatagc cgttggtaca aagcggggat ctgacgagct tttctctact tgtgtcacta
     1021 acggaccgtt tatcatgagc agcaactcgg cttctgcagc aaacggaaat gacagcaaga
     1081 agttcaaagg tgacagccga agtgcaggcg tcccctctag agtgatccac atccggaagc
     1141 tccccatcga cgtcacggag ggggaagtca tctccctggg gctgcccttt gggaaggtca
     1201 ccaacctcct gatgctgaag gggaaaaacc aggccttcat cgagatgaac acggaggagg
     1261 ctgccaacac catggtgaac tactacacct cggtgacccc tgtgctgcgc ggccagccca
     1321 tctacatcca gttctctaac cacaaggagc tgaagaccga cagctctccc aaccaggcgc
     1381 gggcccaggc ggccctgcag gcggtgaact cggtccagtc ggggaacctg gccttggctg
     1441 cctcggcggc ggccgtggac gcagggatgg cgatggccgg gcagagccct gtgctcagga
     1501 tcatcgtgga gaacctcttc taccctgtga ccctggatgt gctgcaccag attttctcca
     1561 agttcggcac agtgttgaag atcatcacct tcaccaagaa caaccagttc caggccctgc
     1621 tgcagtatgc ggaccccgtg agcgcccagc acgccaagct gtcgctggac gggcagaaca
     1681 tctacaacgc ctgctgcacg ctgcgcatcg acttttccaa gctcaccagc ctcaacgtca
     1741 agtacaacaa tgacaagagc cgtgactaca cacgcccaga cctgccttcc ggggacagcc
     1801 agccctcgct ggaccagacc atggccgcgg ccttcggtgc acctggtata atctcagcct
     1861 ctccgtatgc aggagctggt ttccctccca cctttgccat tcctcaagct gcaggccttt
     1921 ccgttccgaa cgtccacggc gccctggccc ccctggccat cccctcggcg gcggcggcag
     1981 ctgcggcggc aggtcggatc gccatcccgg gcctggcggg ggcaggaaat tctgtattgc
     2041 tggtcagcaa cctcaaccca gagagagtca caccccaaag cctctttatt cttttcggcg
     2101 tctacggtga cgtgcagcgc gtgaagatcc tgttcaataa gaaggagaac gccctagtgc
     2161 agatggcgga cggcaaccag gcccagctgg ccatgagcca cctgaacggg cacaagctgc
     2221 acgggaagcc catccgcatc acgctctcga agcaccagaa cgtgcagctg ccccgcgagg
     2281 gccaggagga ccagggcctg accaaggact acggcaactc acccctgcac cgcttcaaga
     2341 agccgggctc caagaacttc cagaacatat tcccgccctc ggccacgctg cacctctcca
     2401 acatcccgcc ctcagtctcc gaggaggatc tcaaggtcct gttttccagc aatgggggcg
     2461 tcgtcaaagg attcaagttc ttccagaagg accgcaagat ggcactgatc cagatgggct
     2521 ccgtggagga ggcggtccag gccctcattg acctgcacaa ccacgacctc ggggagaacc
     2581 accacctgcg ggtctccttc tccaagtcca ccatctagct cgagcggccg catcgtgact
     2641 gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     2701 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     2761 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     2821 gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     2881 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     2941 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3001 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3061 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3121 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3181 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3241 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     3301 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3361 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3421 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3481 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3541 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     3601 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3661 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3721 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3781 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3841 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3901 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3961 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4021 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4081 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4141 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4201 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4261 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4321 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4381 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4441 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4501 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4561 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4621 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4681 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4741 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4801 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     4861 ttctccttac gcatctgtgc ggtatttcac accgcataaa ttccgacacc atcgaatggc
     4921 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
     4981 atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg
     5041 tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag
     5101 cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac
     5161 agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg
     5221 tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag
     5281 aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca
     5341 gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct
     5401 gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta
     5461 ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc
     5521 agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg
     5581 gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact
     5641 ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca
     5701 ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt
     5761 ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct
     5821 catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca
     5881 gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc
     5941 ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc
     6001 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     6061 agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcacccca ggctttacac
     6121 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     6181 aacagctatg accatgatta cggattcact ggccgtcgtt ttacaacgtc gtgactggga
     6241 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     6301 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     6361 atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc tggagtgcga
     6421 tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc
     6481 gcccatctac accaacgtga cctatcccat tacggtcaat ccgccgtttg ttcccacgga
     6541 gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc tacaggaagg
     6601 ccagacgcga attatttttg atggcgttgg aatt
//
