LOCUS       sequence-335776-        5945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial experssion of a GST-tagged MEG-3 Cterminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5945)
  AUTHORS   Schmidt H, Putnam A, Rasoloson D, Seydoux G
  TITLE     Coordination of RNA and protein condensation by the P granule 
            protein MEG-3
  JOURNAL   bioRxiv 2020.10.15.340570
REFERENCE   2  (bases 1 to 5945)
  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: "bioRxiv 
            2020.10.15.340570"
COMMENT     Sequence Label: pDC25
FEATURES             Location/Qualifiers
     source          1..5945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(22..27,28..43,44..50)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    58..74
                     /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             97..750
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     708..730
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             757..780
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(1834..1856)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2001..2019)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2087..2191
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2192..2260,2261..3052)
                     /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(2410..2429)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3223..3811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3712..3731
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3965..3982
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4055..4132
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4133..5215
                     /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(4152..4171)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5264..5269,5270..5287,5288..5294)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5302..5318
                     /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     5326..5342
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5326..5342
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5338..5865
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5358..5375)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5358..5374)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5367..5389)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg tgtggaattg
       61 tgagcggata acaatttcac acaggaaaca gtattcatgt cccctatact aggttattgg
      121 aaaattaagg gccttgtgca acccactcga cttcttttgg aatatcttga agaaaaatat
      181 gaagagcatt tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg
      241 ggtttggagt ttcccaatct tccttattat attgatggtg atgttaaatt aacacagtct
      301 atggccatca tacgttatat agctgacaag cacaacatgt tgggtggttg tccaaaagag
      361 cgtgcagaga tttcaatgct tgaaggagcg gttttggata ttagatacgg tgtttcgaga
      421 attgcatata gtaaagactt tgaaactctc aaagttgatt ttcttagcaa gctacctgaa
      481 atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat atttaaatgg tgatcatgta
      541 acccatcctg acttcatgtt gtatgacgct cttgatgttg ttttatacat ggacccaatg
      601 tgcctggatg cgttcccaaa attagtttgt tttaaaaaac gtattgaagc tatcccacaa
      661 attgataagt acttgaaatc cagcaagtat atagcatggc ctttgcaggg ctggcaagcc
      721 acgtttggtg gtggcgacca tcctccaaaa tcggatctgg aagttctgtt ccaggggccc
      781 ctgggatccc cggaattccc gggtcgacac atgtcgtcaa tcatcccaac tccaaaggct
      841 ttgatcggag aaagatgtct gacatcatcc agcaagtctt caaagttgaa caagtcactt
      901 ggcgtggttg attctaaagc tacaaaatca ccaatgtatt ccgtcacagt ttctggaaaa
      961 gaaacagctt caggaaagcg aattgctcaa aagttgactc caaaggttgt ggcgctggaa
     1021 agctcttaca ttaccggaat tccagtttcc acggattgta atggttgccc aactccgaag
     1081 agaagtggaa tcaattgtga aattcgtgca gctgaagtat acaatcaagc tgggaaatgg
     1141 cccttcgaaa tcaccagtga tccggctcca cttccttgtg aatcagccga tcggatcgaa
     1201 tatccttctc aagattgcac tcaagatcca gcttcaacct cgccaccacc tcgcatttct
     1261 gaaagcttga cagcattcct ggaagctcag caagacttta acgactacat tgatactaac
     1321 tacaaagaaa agacacaact cctgaaagtg aatttgaata ttcacgggat gtcacctgaa
     1381 agatggctgt acttgaatta tttttgcacc gagacaattc caagattgga tggtccttat
     1441 gccgatgatc caagagttcc acctgtgcga aatatgttca gaaaatggtt cctccgtttt
     1501 gctgaagcct gcttaggaaa tcctcaccag cttgcagtta tgcaagagat tgctgccacg
     1561 tttgttcagg ctcggctgga cgatacgagc agctccacag actccaccaa tatgctctac
     1621 atgttgtgga aggagtgtat cggacagaag aatatcatcg caattgctga tgcgtgttta
     1681 cttgcacatc ttcggaaatc ggatcctatc aaatacttga atgtaaaacg ggattggctg
     1741 gaatctattt ttgacccacc cagagatcaa tgagcggccg catcgtgact gactgacgat
     1801 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     1861 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     1921 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     1981 taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     2041 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2101 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2161 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     2221 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2281 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     2341 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     2401 taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg
     2461 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2521 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2581 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2641 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2701 cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa
     2761 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     2821 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2881 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2941 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     3001 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3061 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     3121 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     3181 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3241 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3301 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3361 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3421 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3481 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3541 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3601 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3661 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3721 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3781 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3841 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3901 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3961 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     4021 gcatctgtgc ggtatttcac accgcataaa ttccgacacc atcgaatggc gcaaaacctt
     4081 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
     4141 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
     4201 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
     4261 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
     4321 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
     4381 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
     4441 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
     4501 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
     4561 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
     4621 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
     4681 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
     4741 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
     4801 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
     4861 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
     4921 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
     4981 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
     5041 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
     5101 ggtgaaaaga aaaaccacgc tagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     5161 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     5221 acgcaattaa tgtaagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     5281 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     5341 accatgatta cggattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     5401 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     5461 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt
     5521 gcctggtttc cggcaccaga agcggtgccg gaaagctggc tggagtgcga tcttcctgag
     5581 gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc gcccatctac
     5641 accaacgtga cctatcccat tacggtcaat ccgccgtttg ttcccacgga gaatccgacg
     5701 ggttgttact cgctcacatt taatgttgat gaaagctggc tacaggaagg ccagacgcga
     5761 attatttttg atggcgttgg aattagctta tcgactgcac ggtgcaccaa tgcttctggc
     5821 gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc
     5881 gtgtcgctca aggcgcactc ccgttctgga taatgttttt tgcgccgaca tcataacggt
     5941 tctgg
//
