LOCUS       sequence-437155-        6286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6286)
  AUTHORS   Wollert T, Wunder C, Lippincott-Schwartz J, Hurley JH
  TITLE     Membrane scission by the ESCRT-III complex.
  JOURNAL   Nature. 2009 Mar 12. 458(7235):172-7.
  PUBMED    19234443
REFERENCE   2  (bases 1 to 6286)
  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: "Nature. 
            2009 Mar 12. 458(7235):172-7."
COMMENT     Sequence Label: pGST-Vps4
FEATURES             Location/Qualifiers
     source          1..6286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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             76..729
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     687..709
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             742..762
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2154..2176)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2321..2339)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2407..2511
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2512..2580,2581..3372)
                     /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(2730..2749)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3543..4131
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4032..4051
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4285..4302
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4375..4452
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4453..5535
                     /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(4472..4491)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5584..5589,5590..5607,5608..5614)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5622..5638
                     /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     5646..5662
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5646..5662
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5658..6185
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5678..5695)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5678..5694)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5687..5709)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      121 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      181 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      241 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      301 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      361 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      421 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      481 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
      541 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
      601 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
      661 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
      721 cctccaaaaa tcgatacgac cgaaaacctg tattttcagg gcgccatggg atccatgagc
      781 acgggagatt ttttaactaa gggaatagaa ctggttcaaa aggccattga cctggacaca
      841 gccacgcagt atgaggaggc gtatacagca tactataacg gattagatta tttgatgttg
      901 gctttgaaat acgagaaaaa ccctaagtcg aaggatttga taagagcgaa gtttacagag
      961 tatctaaacc gtgcagagca gctgaaaaag cacttggaaa gtgaagaggc aaacgcggca
     1021 aagaaatctc ctagcgctgg tagtggttcc aacggtggta ataagaaaat ctctcaggaa
     1081 gagggtgagg ataatggtgg agaagacaac aaaaagctaa gaggtgcctt atcgagtgct
     1141 attttgtctg agaagcccaa tgttaagtgg gaagacgtgg ctggactgga aggagccaaa
     1201 gaggccctta aagaagccgt tattttacct gtcaagttcc cacatttgtt caaaggtaac
     1261 cgtaagccca cttcgggaat cttattgtac ggaccaccag gtacaggtaa atcatatttg
     1321 gcaaaagcag tggccaccga ggccaattct acctttttca gtgttagttc cagtgatttg
     1381 gtttctaagt ggatgggtga atctgaaaaa cttgtcaaac agttatttgc gatggcaaga
     1441 gaaaataaac cttctattat ttttatagat gaagtggatg cgctaacagg taccagagga
     1501 gaaggagaga gcgaagcaag tagaagaatt aaaacagaat tattggtaca aatgaatggt
     1561 gttgggaatg actcccaagg tgttttggtc cttggcgcaa caaatatccc atggcaactg
     1621 gacagtgcca tcaggagaag atttgaaaga agaatatata taccattacc agatttggcg
     1681 gcaagaacca ccatgttcga gattaacgtc ggcgacactc catgcgtatt gaccaaggaa
     1741 gattatagaa ctttaggtgc aatgactgaa ggttattccg ggagtgatat tgctgtagtg
     1801 gtgaaagatg cgctaatgca accgataaga aaaattcaaa gcgcgacaca tttcaaagat
     1861 gtctctacgg aggacgacga aacaagaaaa ctaacaccat gctctccagg tgatgatggc
     1921 gccatcgaga tgagttggac ggatattgaa gctgatgaat taaaagaacc agatctcacc
     1981 ataaaggatt tcttaaaggc tatcaaatcg acgagaccta ccgtgaatga ggatgacttg
     2041 ctgaagcaag aacagttcac tagagatttt ggtcaagaag gtaactagct cgagcggccg
     2101 catcgtgact gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     2161 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     2221 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     2281 cgtagcgata gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt
     2341 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     2401 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2461 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     2521 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2581 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2641 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2701 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     2761 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2821 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2881 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2941 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3001 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     3061 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3121 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3181 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3241 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3301 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3361 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3421 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3481 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3541 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3601 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3661 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     3721 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3781 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3841 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3901 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3961 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4021 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4081 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4141 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4201 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4261 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     4321 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataaa ttccgacacc
     4381 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     4441 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
     4501 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     4561 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     4621 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     4681 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     4741 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     4801 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     4861 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     4921 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     4981 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     5041 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     5101 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     5161 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     5221 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
     5281 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     5341 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     5401 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     5461 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     5521 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcacccca
     5581 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     5641 tcacacagga aacagctatg accatgatta cggattcact ggccgtcgtt ttacaacgtc
     5701 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     5761 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     5821 tgaatggcga atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc
     5881 tggagtgcga tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg
     5941 gttacgatgc gcccatctac accaacgtga cctatcccat tacggtcaat ccgccgtttg
     6001 ttcccacgga gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc
     6061 tacaggaagg ccagacgcga attatttttg atggcgttgg aattagctta tcgactgcac
     6121 ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt
     6181 cgtaaatcac tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt
     6241 tgcgccgaca tcataacggt tctggcaaat attctgaaat gagctg
//
