LOCUS       sequence-218624-        5746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of GST-tagged p37 lacking the N-terminal AA 
            2-81.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5746)
  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 5746)
  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 p37delta2-81
FEATURES             Location/Qualifiers
     source          1..5746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(80..85,86..101,102..108)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    116..132
                     /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             155..808
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     766..788
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             815..838
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1693..1715)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1860..1878)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1946..2050
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2051..2119,2120..2911)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSDKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2269..2288)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3082..3670
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3571..3590
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3824..3841
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3914..3991
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3992..5074
                     /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(4011..4030)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5123..5128,5129..5146,5147..5153)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5161..5177
                     /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     5185..5201
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5185..5201
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5197..5724
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5217..5234)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5217..5233)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5226..5248)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg acatcataac ggttctggca
       61 aatattctga aatgagctgt tgacaattaa tcatcggctc gtataatgtg tggaattgtg
      121 agcggataac aatttcacac aggaaacagt attcatgtcc cctatactag gttattggaa
      181 aattaagggc cttgtgcaac ccactcgact tcttttggaa tatcttgaag aaaaatatga
      241 agagcatttg tatgagcgcg atgaaggtga taaatggcga aacaaaaagt ttgaattggg
      301 tttggagttt cccaatcttc cttattatat tgatggtgat gttaaattaa cacagtctat
      361 ggccatcata cgttatatag ctgacaagca caacatgttg ggtggttgtc caaaagagcg
      421 tgcagagatt tcaatgcttg aaggagcggt tttggatatt agatacggtg tttcgagaat
      481 tgcatatagt aaagactttg aaactctcaa agttgatttt cttagcaagc tacctgaaat
      541 gctgaaaatg ttcgaagatc gtttatgtca taaaacatat ttaaatggtg atcatgtaac
      601 ccatcctgac ttcatgttgt atgacgctct tgatgttgtt ttatacatgg acccaatgtg
      661 cctggatgcg ttcccaaaat tagtttgttt taaaaaacgt attgaagcta tcccacaaat
      721 tgataagtac ttgaaatcca gcaagtatat agcatggcct ttgcagggct ggcaagccac
      781 gtttggtggt ggcgaccatc ctccaaaatc ggatctggaa gttctgttcc aggggcccct
      841 gggatccatg gggaaaattg tgaatgaact tttcaaagag gcaagggaac atggagctgt
      901 ccctctaaac gaagccacaa gatcttcaag ggaagacaaa actaagtcat ttacaggtgg
      961 aggatacaga ttggggaatt ccttctacaa gcgatcagag tatatctatg gggaaaatca
     1021 gctccaagat gttcaggttt tactgaagct gtggcgcaat ggtttcagtt tagacgatgg
     1081 agaattgagg ccttacagtg acccaacaaa tgctcagttt ttggaatctg ttaagagagg
     1141 agagactccc ctggagcttc agcgactggt ccatggcgcc caagtgaacc tggatatgga
     1201 ggatcatcaa gaccaagagt acataaaacc caggttgcga ttcaaggctt ttagtggaga
     1261 agggcaaaaa cttggaagcc ttacacctga aatcgtcagt actccttcct ccccagaaga
     1321 agaggataaa tcaatactga atgcagctgt tcttattgat gattcaatgc ccaccacaaa
     1381 aattcagatc aggttggcag atggaagtcg tctcgtacag agattcaata gcacacaccg
     1441 gattctggat gtacgagact ttatagtacg gtcacgtcct gaatttgcga ctactgactt
     1501 tattcttgtg acttcatttc caagtaaaga actaacagat gaaaccgtga cactacaaga
     1561 agcggacatt cttaacactg tgatactcca acagctaaaa tagggatccc cggaattccc
     1621 gggtcgactc gagcggccgc atcgtgactg actgacgatc tgcctcgcgc gtttcggtga
     1681 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     1741 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     1801 cgcagccatg acccagtcac gtagcgatag cggagtgtat aattcttgaa gacgaaaggg
     1861 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     1921 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1981 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2041 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2101 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2161 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcgataaga tccttgagag
     2221 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2281 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2341 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2401 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2461 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2521 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2581 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2641 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2701 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2761 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2821 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2881 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2941 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3001 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3061 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3121 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3181 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     3241 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3301 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3361 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3421 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3481 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3541 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3601 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3661 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3721 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3781 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3841 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3901 ccgcataaat tccgacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg
     3961 cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc
     4021 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
     4081 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
     4141 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
     4201 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
     4261 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
     4321 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
     4381 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
     4441 ctctgaccag acacccatca acagtattat tttctcccat gaagacggta cgcgactggg
     4501 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
     4561 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
     4621 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
     4681 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
     4741 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt
     4801 agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa
     4861 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
     4921 ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccacgct
     4981 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     5041 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc
     5101 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     5161 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac ggattcactg
     5221 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     5281 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     5341 tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa
     5401 gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc
     5461 tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtgac ctatcccatt
     5521 acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt
     5581 aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttgga
     5641 attagcttat cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc atcggaagct
     5701 gtggtatggc tgtgcaggtc gtaaatcact gcataattcg tgtcgc
//
