LOCUS       sequence-218617-        5986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of GST-tagged p37.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5986)
  AUTHORS   Weith M, Seiler J, van den Boom J, Kracht M, Hulsmann J, Primorac I,
            Del Pino Garcia J, Kaschani F, Kaiser M, Musacchio A, Bollen M, 
            Meyer H
  TITLE     Ubiquitin-Independent Disassembly by a p97 AAA-ATPase Complex Drives
            PP1 Holoenzyme Formation.
  JOURNAL   Mol Cell. 2018 Oct 11. pii: S1097-2765(18)30791-3. doi: 
            10.1016/j.molcel.2018.09.020.
  PUBMED    30344098
REFERENCE   2  (bases 1 to 5986)
  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"; journalName: "Mol Cell. 
            2018 Oct 11. pii: S1097-2765(18)30791-3. doi: 
            10.1016/j.molcel.2018.09.020."
COMMENT     Sequence Label: pGEX-6P-1 p37
FEATURES             Location/Qualifiers
     source          1..5986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(32..37,38..53,54..60)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    68..84
                     /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             107..760
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     718..740
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             767..790
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1885..1907)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2052..2070)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2138..2242
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2243..2311,2312..3103)
                     /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(2461..2480)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3274..3862
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3763..3782
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4016..4033
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4106..4183
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4184..5266
                     /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(4203..4222)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5315..5320,5321..5338,5339..5345)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5353..5369
                     /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     5377..5393
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5377..5393
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5389..5916
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5409..5426)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5409..5425)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5418..5440)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg
       61 tgtggaattg tgagcggata acaatttcac acaggaaaca gtattcatgt cccctatact
      121 aggttattgg aaaattaagg gccttgtgca acccactcga cttcttttgg aatatcttga
      181 agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa
      241 gtttgaattg ggtttggagt ttcccaatct tccttattat attgatggtg atgttaaatt
      301 aacacagtct atggccatca tacgttatat agctgacaag cacaacatgt tgggtggttg
      361 tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg gttttggata ttagatacgg
      421 tgtttcgaga attgcatata gtaaagactt tgaaactctc aaagttgatt ttcttagcaa
      481 gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat atttaaatgg
      541 tgatcatgta acccatcctg acttcatgtt gtatgacgct cttgatgttg ttttatacat
      601 ggacccaatg tgcctggatg cgttcccaaa attagtttgt tttaaaaaac gtattgaagc
      661 tatcccacaa attgataagt acttgaaatc cagcaagtat atagcatggc ctttgcaggg
      721 ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa tcggatctgg aagttctgtt
      781 ccaggggccc ctgggatcca tggcggaggg tggccgtgcg gagccggagg agcaggagcg
      841 tgggtcctcc aggccacggc ccccaagtgc gcgggactta cagttggcct tggcagaatt
      901 atatgaagat gaaatgaaat gcaagtcttc caagcctgac agatccacac ctgcaacctg
      961 taggagccca cgaacaccgc cacatcggct ttattcaggt gatcacaaat atgatggatt
     1021 acatatagtt caacctccaa ctgggaaaat tgtgaatgaa cttttcaaag aggcaaggga
     1081 acatggagct gtccctctaa acgaagccac aagatcttca agggaagaca aaactaagtc
     1141 atttacaggt ggaggataca gattggggaa ttccttctac aagcgatcag agtatatcta
     1201 tggggaaaat cagctccaag atgttcaggt tttactgaag ctgtggcgca atggtttcag
     1261 tttagacgat ggagaattga ggccttacag tgacccaaca aatgctcagt ttttggaatc
     1321 tgttaagaga ggagagactc ccctggagct tcagcgactg gtccatggcg cccaagtgaa
     1381 cctggatatg gaggatcatc aagaccaaga gtacataaaa cccaggttgc gattcaaggc
     1441 ttttagtgga gaagggcaaa aacttggaag ccttacacct gaaatcgtca gtactccttc
     1501 ctccccagaa gaagaggata aatcaatact gaatgcagct gttcttattg atgattcaat
     1561 gcccaccaca aaaattcaga tcaggttggc agatggaagt cgtctcgtac agagattcaa
     1621 tagcacacac cggattctgg atgtacgaga ctttatagta cggtcacgtc ctgaatttgc
     1681 gactactgac tttattcttg tgacttcatt tccaagtaaa gaactaacag atgaaaccgt
     1741 gacactacaa gaagcggaca ttcttaacac tgtgatactc caacagctaa aatagggatc
     1801 cccggaattc ccgggtcgac tcgagcggcc gcatcgtgac tgactgacga tctgcctcgc
     1861 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     1921 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     1981 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt ataattcttg
     2041 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     2101 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2161 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2221 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2281 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2341 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2401 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2461 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     2521 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     2581 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     2641 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     2701 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     2761 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     2821 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     2881 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2941 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3001 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3061 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3121 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3181 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3241 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3301 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3361 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3421 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3481 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3541 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3601 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3661 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3721 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3781 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3841 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3901 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3961 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4021 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     4081 cggtatttca caccgcataa attccgacac catcgaatgg cgcaaaacct ttcgcggtat
     4141 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
     4201 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
     4261 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
     4321 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
     4381 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
     4441 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
     4501 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
     4561 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
     4621 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
     4681 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
     4741 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
     4801 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
     4861 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
     4921 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
     4981 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
     5041 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
     5101 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
     5161 aaaaaccacg ctagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5221 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5281 atgtaagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     5341 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     5401 acggattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     5461 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     5521 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt
     5581 ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga ggccgatact
     5641 gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta caccaacgtg
     5701 acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac gggttgttac
     5761 tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg aattattttt
     5821 gatggcgttg gaattagctt atcgactgca cggtgcacca atgcttctgg cgtcaggcag
     5881 ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc
     5941 aaggcgcact cccgttctgg ataatgtttt ttgcgccgac atcata
//
