LOCUS       sequence-218626-        5986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of GST-tagged p37 that is p97 binding deficient
            due to mutations in the SHP box .
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 SHPmut
FEATURES             Location/Qualifiers
     source          1..5986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(47..52,53..68,69..75)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    83..99
                     /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             122..775
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     733..755
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             782..805
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1900..1922)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2067..2085)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2153..2257
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2258..2326,2327..3118)
                     /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(2476..2495)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3289..3877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3778..3797
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4031..4048
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4121..4198
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4199..5281
                     /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(4218..4237)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5330..5335,5336..5353,5354..5360)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5368..5384
                     /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     5392..5408
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5392..5408
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5404..5931
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5424..5441)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5424..5440)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5433..5455)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tgcgccgaca tcataacggt tctggcaaat attctgaaat gagctgttga caattaatca
       61 tcggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagtatt
      121 catgtcccct atactaggtt attggaaaat taagggcctt gtgcaaccca ctcgacttct
      181 tttggaatat cttgaagaaa aatatgaaga gcatttgtat gagcgcgatg aaggtgataa
      241 atggcgaaac aaaaagtttg aattgggttt ggagtttccc aatcttcctt attatattga
      301 tggtgatgtt aaattaacac agtctatggc catcatacgt tatatagctg acaagcacaa
      361 catgttgggt ggttgtccaa aagagcgtgc agagatttca atgcttgaag gagcggtttt
      421 ggatattaga tacggtgttt cgagaattgc atatagtaaa gactttgaaa ctctcaaagt
      481 tgattttctt agcaagctac ctgaaatgct gaaaatgttc gaagatcgtt tatgtcataa
      541 aacatattta aatggtgatc atgtaaccca tcctgacttc atgttgtatg acgctcttga
      601 tgttgtttta tacatggacc caatgtgcct ggatgcgttc ccaaaattag tttgttttaa
      661 aaaacgtatt gaagctatcc cacaaattga taagtacttg aaatccagca agtatatagc
      721 atggcctttg cagggctggc aagccacgtt tggtggtggc gaccatcctc caaaatcgga
      781 tctggaagtt ctgttccagg ggcccctggg atccatggcg gagggtggcc gtgcggagcc
      841 ggaggagcag gagcgtgggt cctccaggcc acggccccca agtgcgcggg acttacagtt
      901 ggccttggca gaattatatg aagatgaaat gaaatgcaag tcttccaagc ctgacagatc
      961 cacacctgca acctgtagga gcccacgaac accgccacat cggctttatt caggtgatca
     1021 caaatatgat ggattacata tagttcaacc tccaactggg aaaattgtga atgaactttt
     1081 caaagaggca agggaacatg gagctgtccc tctaaacgaa gccacaagat cttcaaggga
     1141 agacaaaact aagtcattta caggtggagg atacagattg gggaattcct tctacaagcg
     1201 atcagagtat atctatgggg aaaatcagct ccaagatgtt caggttttac tgaagctgtg
     1261 gcgcaatggt ttcagtttag acgatggaga attgaggcct tacagtgacc caacaaatgc
     1321 tcagtttttg gaatctgtta agagaggaga gactcccctg gagcttcagc gactggtcca
     1381 tggcgcccaa gtgaacctgg atatggagga tcatcaagac caagagtaca taaaacccag
     1441 gttgcgattc aaggctgcta gtggagcagg ggcggcagct ggaagcctta cacctgaaat
     1501 cgtcagtact ccttcctccc cagaagaaga ggataaatca atactgaatg cagctgttct
     1561 tattgatgat tcaatgccca ccacaaaaat tcagatcagg ttggcagatg gaagtcgtct
     1621 cgtacagaga ttcaatagca cacaccggat tctggatgta cgagacttta tagtacggtc
     1681 acgtcctgaa tttgcgacta ctgactttat tcttgtgact tcatttccaa gtaaagaact
     1741 aacagatgaa accgtgacac tacaagaagc ggacattctt aacactgtga tactccaaca
     1801 gctaaaatag ggatccccgg aattcccggg tcgactcgag cggccgcatc gtgactgact
     1861 gacgatctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     1921 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     1981 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
     2041 agtgtataat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     2101 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     2161 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2221 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     2281 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     2341 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     2401 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     2461 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     2521 actcggtcgt cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     2581 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     2641 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     2701 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     2761 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     2821 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     2881 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     2941 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     3001 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     3061 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     3121 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     3181 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     3241 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3301 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3361 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3421 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3481 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3541 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3601 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3661 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3721 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3781 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3841 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3901 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3961 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     4021 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     4081 ccttacgcat ctgtgcggta tttcacaccg cataaattcc gacaccatcg aatggcgcaa
     4141 aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt
     4201 gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc
     4261 ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc
     4321 gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc
     4381 gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc
     4441 ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg
     4501 aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg
     4561 gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac
     4621 taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt
     4681 ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca
     4741 aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg
     4801 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag
     4861 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc
     4921 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg
     4981 gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg
     5041 ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
     5101 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt
     5161 ctcactggtg aaaagaaaaa ccacgctagc gcccaatacg caaaccgcct ctccccgcgc
     5221 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     5281 agcgcaacgc aattaatgta agttagctca ctcattaggc accccaggct ttacacttta
     5341 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     5401 gctatgacca tgattacgga ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     5461 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     5521 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     5581 cgctttgcct ggtttccggc accagaagcg gtgccggaaa gctggctgga gtgcgatctt
     5641 cctgaggccg atactgtcgt cgtcccctca aactggcaga tgcacggtta cgatgcgccc
     5701 atctacacca acgtgaccta tcccattacg gtcaatccgc cgtttgttcc cacggagaat
     5761 ccgacgggtt gttactcgct cacatttaat gttgatgaaa gctggctaca ggaaggccag
     5821 acgcgaatta tttttgatgg cgttggaatt agcttatcga ctgcacggtg caccaatgct
     5881 tctggcgtca ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta aatcactgca
     5941 taattcgtgt cgctcaaggc gcactcccgt tctggataat gttttt
//
