LOCUS       sequence-403788-        7601 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GST-tagged MVP delta607-623aa in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7601)
  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 7601)
  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 delta607-623aa
FEATURES             Location/Qualifiers
     source          1..7601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(212..217,218..233,234..240)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    248..264
                     /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             287..940
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     898..920
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             947..970
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3680..3702)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3847..3865)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3933..4037
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4038..4106,4107..4898)
                     /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(4256..4275)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5069..5657
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5558..5577
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5811..5828
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5901..5978
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5979..7061
                     /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(5998..6017)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(7110..7115,7116..7133,7134..7140)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7148..7164
                     /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     7172..7188
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7172..7188
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(7184..7601,1..110)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(7204..7221)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7204..7220)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7213..7235)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cgcgaattat ttttgatggc gttggaatta gcttatcgac tgcacggtgc accaatgctt
       61 ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat
      121 aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata
      181 acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg
      241 tgtggaattg tgagcggata acaatttcac acaggaaaca gtattcatgt cccctatact
      301 aggttattgg aaaattaagg gccttgtgca acccactcga cttcttttgg aatatcttga
      361 agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa
      421 gtttgaattg ggtttggagt ttcccaatct tccttattat attgatggtg atgttaaatt
      481 aacacagtct atggccatca tacgttatat agctgacaag cacaacatgt tgggtggttg
      541 tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg gttttggata ttagatacgg
      601 tgtttcgaga attgcatata gtaaagactt tgaaactctc aaagttgatt ttcttagcaa
      661 gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat atttaaatgg
      721 tgatcatgta acccatcctg acttcatgtt gtatgacgct cttgatgttg ttttatacat
      781 ggacccaatg tgcctggatg cgttcccaaa attagtttgt tttaaaaaac gtattgaagc
      841 tatcccacaa attgataagt acttgaaatc cagcaagtat atagcatggc ctttgcaggg
      901 ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa tcggatctgg aagttctgtt
      961 ccaggggccc ctgggatccc cggaattcat ggcaactgaa gagttcatca tccgcatccc
     1021 cccataccac tatatccatg tgctggacca gaacagcaac gtgtcccgtg tggaggtcgg
     1081 gccaaagacc tacatccggc aggacaatga gagggtactg tttgccccca tgcgcatggt
     1141 gaccgtcccc ccacgtcact actgcacagt ggccaaccct gtgtctcggg atgcccaggg
     1201 cttggtgctg tttgatgtca cagggcaagt tcggcttcgc cacgctgacc tcgagatccg
     1261 gctggcccag gaccccttcc ccctgtaccc aggggaggtg ctggaaaagg acatcacacc
     1321 cctgcaggtg gttctgccca acactgccct ccatctaaag gcgctgcttg attttgagga
     1381 taaagatgga gacaaggtgg tggcaggaga tgagtggctt ttcgagggac ctggcacgta
     1441 catcccccgg aaggaagtgg aggtcgtgga gatcattcag gccaccatca tcaggcagaa
     1501 ccaggctctg cggctcaggg cccgcaagga gtgctgggac cgggacggca aggagagggt
     1561 gacaggggaa gaatggctgg tcaccacagt aggggcgtac ctcccagcgg tgtttgagga
     1621 ggttctggat ttggtggacg ccgtcatcct tacggaaaag acagccctgc acctccgggc
     1681 tcggcggaac ttccgggact tcaggggagt gtcccgccgc actggggagg agtggctggt
     1741 aacagtgcag gacacagagg cccacgtgcc agatgtccac gaggaggtgc tgggggttgt
     1801 gcccatcacc accctgggcc cccacaacta ctgcgtgatt ctcgaccctg tcggaccgga
     1861 tggcaagaat cagctggggc agaagcgcgt ggtcaaggga gagaagtctt ttttcctcca
     1921 gccaggagag cagctggaac aaggcatcca ggatgtgtat gtgctgtcgg agcagcaggg
     1981 gctgctgctg agggccctgc agcccctgga ggagggggag gatgaggaga aggtctcaca
     2041 ccaggctggg gaccactggc tcatccgcgg acccctggag tatgtgccat ctgccaaagt
     2101 ggaggtggtg gaggagcgcc aggccatccc tctagacgag aacgagggca tctatgtgca
     2161 ggatgtcaag accggaaagg tgcgcgctgt gattggaagc acctacatgc tgacccagga
     2221 cgaagtcctg tgggagaaag agctgcctcc cggggtggag gagctgctga acaaggggca
     2281 ggaccctctg gcagacaggg gtgagaagga cacagctaag agcctccagc ccttggcgcc
     2341 ccggaacaag acccgtgtgg tcagctaccg cgtgccccac aacgctgcgg tgcaggtgta
     2401 cgactaccga gagaagcgag cccgcgtggt cttcgggcct gagctggtgt cgctgggtcc
     2461 tgaggagcag ttcacagtgt tgtccctctc agctgggcgg cccaagcgtc cccatgcccg
     2521 ccgtgcgctc tgcctgctgc tggggcctga cttcttcaca gacgtcatca ccatcgaaac
     2581 ggcggatcat gccaggctgc aactgcagct ggcctacaac tggcactttg aggtgaatga
     2641 ccggaaggac ccccaagaga cggccaagct cttttcagtg ccagactttg taggtgatgc
     2701 ctgcaaagcc atcgcatccc gggtgcgggg ggccgtggcc tctgtcactt tcgatgactt
     2761 ccataagaac tcagcccgca tcattcgcac tgctgtcttt ggctttgacc aggctgtctt
     2821 cccccaaaac gggctggtgg tcagcagtgt ggacgtgcag tcagtggagc ctgtggatca
     2881 gaggacccgg gacgccctgc aacgcagcgt ccagctggcc atcgagatca ccaccaactc
     2941 ccaggaagcg gcggccaagc atgaggctca gagactggag caggaagccc gcggccggct
     3001 tgagcggcag aagatcctgg accagtcaga agccgagaaa gctcgcaagg aacttttgga
     3061 gctggaggct ctgagcatgg ccgtggagag caccgggact gccaaggcgg aggccgagtc
     3121 ccgtgcggag gcagcccgga ttgagggaga agggtccgtg ctgcaggcca agctaaaagc
     3181 acaggccttg gccattgaaa cggaggctga gctccagagg gtccagaagg tccgagagct
     3241 ggaactggtc tatgcccggg cccagctgga gctggaggtg agcaaggctc agcagctggc
     3301 tgaggtggag gtgaagaagt tcaagcagat gacagaggcc ataggcccca gcaccatcag
     3361 ggaccttgct gtggctgggc ctgagatgca ggtaaaactg ctccagtccc tgggcctgaa
     3421 atcaaccctc atcaccgatg gctccactcc catcaacctc ttcaacacag cctttgggct
     3481 gctggggatg gggcccgagg gtcagcccct gggcagaagg gtggccagtg ggcccagccc
     3541 tggggagggg atatcccccc agtctgctca ggcccctcaa gctcctggag acaaccacgt
     3601 ggtgcctgta ctgcgctaag cggccgcatc gtgactgact gacgatctgc ctcgcgcgtt
     3661 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     3721 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     3781 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtataat tcttgaagac
     3841 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     3901 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3961 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4021 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4081 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4141 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4201 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4261 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     4321 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4381 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4441 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4501 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4561 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4621 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4681 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4741 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4801 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4861 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4921 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4981 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5041 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5101 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5161 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     5221 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5281 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5341 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5401 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5461 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5521 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5581 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5641 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5701 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5761 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     5821 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     5881 tttcacaccg cataaattcc gacaccatcg aatggcgcaa aacctttcgc ggtatggcat
     5941 gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg
     6001 atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca
     6061 gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca
     6121 ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca
     6181 cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg
     6241 atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta
     6301 aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc
     6361 tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc
     6421 ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc
     6481 gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc
     6541 cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca
     6601 atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac
     6661 aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc
     6721 agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata
     6781 tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca
     6841 ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct
     6901 ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa
     6961 ccacgctagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     7021 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta
     7081 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     7141 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgga
     7201 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     7261 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     7321 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc
     7381 accagaagcg gtgccggaaa gctggctgga gtgcgatctt cctgaggccg atactgtcgt
     7441 cgtcccctca aactggcaga tgcacggtta cgatgcgccc atctacacca acgtgaccta
     7501 tcccattacg gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt gttactcgct
     7561 cacatttaat gttgatgaaa gctggctaca ggaaggccag a
//
