LOCUS       sequence-403728-        7652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GST-tagged MVP in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7652)
  AUTHORS   Kurusu R, Fujimoto Y, Morishita H, Noshiro D, Takada S, Yamano K, 
            Tanaka H, Arai R, Kageyama S, Funakoshi T, Komatsu-Hirota S, Taka H,
            Kazuno S, Miura Y, Koike M, Wakai T, Waguri S, Noda NN, Komatsu M
  TITLE     Integrated proteomics identifies p62-dependent selective autophagy 
            of the supramolecular vault complex.
  JOURNAL   Dev Cell. 2023 May 9:S1534-5807(23)00191-0. doi: 
            10.1016/j.devcel.2023.04.015.
  PUBMED    37192622
REFERENCE   2  (bases 1 to 7652)
  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: "Dev Cell. 
            2023 May 9:S1534-5807(23)00191-0. doi: 
            10.1016/j.devcel.2023.04.015."
COMMENT     Sequence Label: pGEX6P-1-MVP
FEATURES             Location/Qualifiers
     source          1..7652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(214..219,220..235,236..242)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    250..266
                     /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             289..942
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     900..922
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             949..972
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3733..3755)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3900..3918)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3986..4090
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4091..4159,4160..4951)
                     /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(4309..4328)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5122..5710
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5611..5630
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5864..5881
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5954..6031
                     /gene="lacI"
                     /label=lacI promoter
     CDS             6032..7114
                     /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(6051..6070)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(7163..7168,7169..7186,7187..7193)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7201..7217
                     /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     7225..7241
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7225..7241
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(7237..7652,1..112)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(7257..7274)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7257..7273)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7266..7288)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gacgcgaatt atttttgatg gcgttggaat tagcttatcg actgcacggt gcaccaatgc
       61 ttctggcgtc aggcagccat cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc
      121 ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatca
      181 taacggttct ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcg gctcgtataa
      241 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagtattcat gtcccctata
      301 ctaggttatt ggaaaattaa gggccttgtg caacccactc gacttctttt ggaatatctt
      361 gaagaaaaat atgaagagca tttgtatgag cgcgatgaag gtgataaatg gcgaaacaaa
      421 aagtttgaat tgggtttgga gtttcccaat cttccttatt atattgatgg tgatgttaaa
      481 ttaacacagt ctatggccat catacgttat atagctgaca agcacaacat gttgggtggt
      541 tgtccaaaag agcgtgcaga gatttcaatg cttgaaggag cggttttgga tattagatac
      601 ggtgtttcga gaattgcata tagtaaagac tttgaaactc tcaaagttga ttttcttagc
      661 aagctacctg aaatgctgaa aatgttcgaa gatcgtttat gtcataaaac atatttaaat
      721 ggtgatcatg taacccatcc tgacttcatg ttgtatgacg ctcttgatgt tgttttatac
      781 atggacccaa tgtgcctgga tgcgttccca aaattagttt gttttaaaaa acgtattgaa
      841 gctatcccac aaattgataa gtacttgaaa tccagcaagt atatagcatg gcctttgcag
      901 ggctggcaag ccacgtttgg tggtggcgac catcctccaa aatcggatct ggaagttctg
      961 ttccaggggc ccctgggatc cccggaattc atggcaactg aagagttcat catccgcatc
     1021 cccccatacc actatatcca tgtgctggac cagaacagca acgtgtcccg tgtggaggtc
     1081 gggccaaaga cctacatccg gcaggacaat gagagggtac tgtttgcccc catgcgcatg
     1141 gtgaccgtcc ccccacgtca ctactgcaca gtggccaacc ctgtgtctcg ggatgcccag
     1201 ggcttggtgc tgtttgatgt cacagggcaa gttcggcttc gccacgctga cctcgagatc
     1261 cggctggccc aggacccctt ccccctgtac ccaggggagg tgctggaaaa ggacatcaca
     1321 cccctgcagg tggttctgcc caacactgcc ctccatctaa aggcgctgct tgattttgag
     1381 gataaagatg gagacaaggt ggtggcagga gatgagtggc ttttcgaggg acctggcacg
     1441 tacatccccc ggaaggaagt ggaggtcgtg gagatcattc aggccaccat catcaggcag
     1501 aaccaggctc tgcggctcag ggcccgcaag gagtgctggg accgggacgg caaggagagg
     1561 gtgacagggg aagaatggct ggtcaccaca gtaggggcgt acctcccagc ggtgtttgag
     1621 gaggttctgg atttggtgga cgccgtcatc cttacggaaa agacagccct gcacctccgg
     1681 gctcggcgga acttccggga cttcagggga gtgtcccgcc gcactgggga ggagtggctg
     1741 gtaacagtgc aggacacaga ggcccacgtg ccagatgtcc acgaggaggt gctgggggtt
     1801 gtgcccatca ccaccctggg cccccacaac tactgcgtga ttctcgaccc tgtcggaccg
     1861 gatggcaaga atcagctggg gcagaagcgc gtggtcaagg gagagaagtc ttttttcctc
     1921 cagccaggag agcagctgga acaaggcatc caggatgtgt atgtgctgtc ggagcagcag
     1981 gggctgctgc tgagggccct gcagcccctg gaggaggggg aggatgagga gaaggtctca
     2041 caccaggctg gggaccactg gctcatccgc ggacccctgg agtatgtgcc atctgccaaa
     2101 gtggaggtgg tggaggagcg ccaggccatc cctctagacg agaacgaggg catctatgtg
     2161 caggatgtca agaccggaaa ggtgcgcgct gtgattggaa gcacctacat gctgacccag
     2221 gacgaagtcc tgtgggagaa agagctgcct cccggggtgg aggagctgct gaacaagggg
     2281 caggaccctc tggcagacag gggtgagaag gacacagcta agagcctcca gcccttggcg
     2341 ccccggaaca agacccgtgt ggtcagctac cgcgtgcccc acaacgctgc ggtgcaggtg
     2401 tacgactacc gagagaagcg agcccgcgtg gtcttcgggc ctgagctggt gtcgctgggt
     2461 cctgaggagc agttcacagt gttgtccctc tcagctgggc ggcccaagcg tccccatgcc
     2521 cgccgtgcgc tctgcctgct gctggggcct gacttcttca cagacgtcat caccatcgaa
     2581 acggcggatc atgccaggct gcaactgcag ctggcctaca actggcactt tgaggtgaat
     2641 gaccggaagg acccccaaga gacggccaag ctcttttcag tgccagactt tgtaggtgat
     2701 gcctgcaaag ccatcgcatc ccgggtgcgg ggggccgtgg cctctgtcac tttcgatgac
     2761 ttccataaga actcagcccg catcattcgc actgctgtct ttggctttga gacctcggaa
     2821 gcgaagggcc ccgatggcat ggccctgccc aggccccggg accaggctgt cttcccccaa
     2881 aacgggctgg tggtcagcag tgtggacgtg cagtcagtgg agcctgtgga tcagaggacc
     2941 cgggacgccc tgcaacgcag cgtccagctg gccatcgaga tcaccaccaa ctcccaggaa
     3001 gcggcggcca agcatgaggc tcagagactg gagcaggaag cccgcggccg gcttgagcgg
     3061 cagaagatcc tggaccagtc agaagccgag aaagctcgca aggaactttt ggagctggag
     3121 gctctgagca tggccgtgga gagcaccggg actgccaagg cggaggccga gtcccgtgcg
     3181 gaggcagccc ggattgaggg agaagggtcc gtgctgcagg ccaagctaaa agcacaggcc
     3241 ttggccattg aaacggaggc tgagctccag agggtccaga aggtccgaga gctggaactg
     3301 gtctatgccc gggcccagct ggagctggag gtgagcaagg ctcagcagct ggctgaggtg
     3361 gaggtgaaga agttcaagca gatgacagag gccataggcc ccagcaccat cagggacctt
     3421 gctgtggctg ggcctgagat gcaggtaaaa ctgctccagt ccctgggcct gaaatcaacc
     3481 ctcatcaccg atggctccac tcccatcaac ctcttcaaca cagcctttgg gctgctgggg
     3541 atggggcccg agggtcagcc cctgggcaga agggtggcca gtgggcccag ccctggggag
     3601 gggatatccc cccagtctgc tcaggcccct caagctcctg gagacaacca cgtggtgcct
     3661 gtactgcgct aagcggccgc atcgtgactg actgacgatc tgcctcgcgc gtttcggtga
     3721 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     3781 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     3841 cgcagccatg acccagtcac gtagcgatag cggagtgtat aattcttgaa gacgaaaggg
     3901 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     3961 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4021 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4081 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4141 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4201 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4261 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4321 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4381 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4441 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4501 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4561 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4621 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4681 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4741 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4801 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4861 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4921 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4981 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5041 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5101 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5161 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5221 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     5281 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5341 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5401 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5461 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5521 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5581 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5641 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5701 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5761 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     5821 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     5881 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     5941 ccgcataaat tccgacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg
     6001 cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc
     6061 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
     6121 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
     6181 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
     6241 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
     6301 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
     6361 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
     6421 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
     6481 ctctgaccag acacccatca acagtattat tttctcccat gaagacggta cgcgactggg
     6541 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
     6601 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
     6661 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
     6721 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
     6781 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt
     6841 agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa
     6901 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
     6961 ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccacgct
     7021 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     7081 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc
     7141 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     7201 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac ggattcactg
     7261 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     7321 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     7381 tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa
     7441 gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc
     7501 tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtgac ctatcccatt
     7561 acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt
     7621 aatgttgatg aaagctggct acaggaaggc ca
//
