LOCUS       sequence-335925-        5977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GST-p37 (human, codon optimized for expression in 
            bacteria).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5977)
  AUTHORS   Kracht M, van den Boom J, Seiler J, Kroning A, Kaschani F, Kaiser M,
            Meyer H
  TITLE     Protein Phosphatase-1 Complex Disassembly by p97 is Initiated 
            through Multivalent Recognition of Catalytic and Regulatory Subunits
            by the p97 SEP-domain Adapters.
  JOURNAL   J Mol Biol. 2020 Nov 20;432(23):6061-6074. doi: 
            10.1016/j.jmb.2020.10.001. Epub 2020 Oct 12.
  PUBMED    33058883
REFERENCE   2  (bases 1 to 5977)
  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: 
            "10.1016/j.jmb.2020.10.001"; journalName: "J Mol Biol"; date: 
            "2020-11-20- 20"; volume: "432"; issue: "23"; pages: "6061-6074"
COMMENT     Sequence Label: pGEX6P1 p37
FEATURES             Location/Qualifiers
     source          1..5977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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             76..729
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     687..709
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             736..759
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1845..1867)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2012..2030)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2098..2202
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2203..2271,2272..3063)
                     /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(2421..2440)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3234..3822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3723..3742
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3976..3993
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4066..4143
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4144..5226
                     /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(4163..4182)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5275..5280,5281..5298,5299..5305)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5313..5329
                     /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     5337..5353
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5337..5353
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5349..5876
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5369..5386)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5369..5385)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5378..5400)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      121 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      181 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      241 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      301 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      361 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      421 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      481 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
      541 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
      601 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
      661 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
      721 cctccaaaat cggatctgga agttctgttc caggggcccc tgggatccat ggcggagggc
      781 ggcggtccag aacctggtga acaagagcgc cgtagctctg gcccgcgccc gccaagcgcc
      841 cgtgatctgc aattggcact ggcggagctg tacgaggatg aggtgaaatg caaaagctcc
      901 aaaagcaacc gccctaaggc gacggtattt aagtctcctc gcacgccgcc gcaacgtttc
      961 tatagcagcg agcacgagta tagcggcctt aacatcgtcc gtccgtctac cggcaaaatc
     1021 gttaatgaac tgttcaaaga agctcgcgaa cacggtgcag ttccactgaa cgaagcaacg
     1081 cgtgcgtctg gcgatgacaa aagcaagagc tttaccggtg gcggttaccg tttgggcagc
     1141 agcttttgta aacgttccga atatatttat ggtgagaacc agcttcagga cgttcagatc
     1201 cttcttaaac tttggagcaa cggctttagc ttggatgatg gcgaacttcg cccttataac
     1261 gagccgacga acgctcaatt cctggaatcc gtcaaacgcg gtgagatccc acttgaactg
     1321 caacgcctgg ttcacggcgg ccaggtaaat cttgacatgg aagaccacca ggatcaagag
     1381 tacatcaagc cgcgcctgcg ctttaaggcc tttagcggcg agggtcagaa gctgggctcc
     1441 ctgacgcctg aaatcgtgag cacccctagc agcccggaag aggaagataa gtccattttg
     1501 aatgcggtag tcttgattga tgatagcgtc ccgaccacca agattcaaat ccgtctggcg
     1561 gatggcagcc gtcttattca acgttttaac agcacccatc gtatcctgga tgtccgcaat
     1621 ttcatcgtcc aaagccgccc ggaatttgct gccctggact ttattcttgt cacctctttc
     1681 ccgaataaag aacttaccga tgagagcctg acccttctgg aagccgatat tttgaatact
     1741 gtgctgctgc aacaactgaa ataagaattc ccgggtcgac tcgagcggcc gcatcgtgac
     1801 tgactgacga tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     1861 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     1921 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     1981 agcggagtgt ataattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     2041 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     2101 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2161 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2221 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     2281 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2341 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     2401 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     2461 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     2521 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     2581 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     2641 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2701 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     2761 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2821 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2881 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2941 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3001 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3061 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3121 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3181 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3241 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3301 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3361 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     3421 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3481 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3541 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3601 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3661 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3721 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3781 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3841 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3901 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3961 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     4021 tttctcctta cgcatctgtg cggtatttca caccgcataa attccgacac catcgaatgg
     4081 cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
     4141 aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
     4201 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
     4261 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
     4321 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
     4381 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
     4441 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
     4501 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
     4561 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
     4621 attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
     4681 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
     4741 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
     4801 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
     4861 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
     4921 tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc
     4981 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
     5041 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
     5101 cccgtctcac tggtgaaaag aaaaaccacg ctagcgccca atacgcaaac cgcctctccc
     5161 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     5221 cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcacccc aggctttaca
     5281 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     5341 aaacagctat gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg
     5401 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     5461 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     5521 aatggcgctt tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg
     5581 atcttcctga ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg
     5641 cgcccatcta caccaacgtg acctatccca ttacggtcaa tccgccgttt gttcccacgg
     5701 agaatccgac gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag
     5761 gccagacgcg aattattttt gatggcgttg gaattagctt atcgactgca cggtgcacca
     5821 atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca
     5881 ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg ataatgtttt ttgcgccgac
     5941 atcataacgg ttctggcaaa tattctgaaa tgagctg
//
