LOCUS       sequence-104086-        6893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of GST-MEG-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6893)
  AUTHORS   Wang JT, Smith J, Chen BC, Schmidt H, Rasoloson D, Paix A, Lambrus 
            BG, Calidas D, Betzig E, Seydoux G
  TITLE     Regulation of RNA granule dynamics by phosphorylation of 
            serine-rich, intrinsically disordered proteins in C. elegans.
  JOURNAL   Elife. 2014 Dec 23;4. doi: 10.7554/eLife.04591.
  PUBMED    25535836
REFERENCE   2  (bases 1 to 6893)
  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: "Elife."; 
            date: "2014-12-23"; volume: "4. doi"; pages: " 10.7554/eLife.04591"
COMMENT     Sequence Label: pJW4.31
FEATURES             Location/Qualifiers
     source          1..6893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2943..2965)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3110..3128)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3196..3300
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3301..3369,3370..4161)
                     /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(3519..3538)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4332..4920
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4821..4840
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5074..5091
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5164..5241
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             5242..6324
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5261..5280)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    6337..6358
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6373..6378,6379..6396,6397..6403)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6411..6427
                     /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     6435..6451
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6435..6451
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6447..6893,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGITLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6467..6484)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6467..6483)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6476..6498)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc ccggaattcc
      961 cgggtcgact catggataat cgtggtcatt tttcttcaaa tgggaatttc ccaccgcaag
     1021 gatatcaccg acgtgagcag tcccaagaag gaatgagaat cgggaatcat cacggaccgt
     1081 tttcttcggg caacatgcgt agtattggag gctcagctca aaaccagcag caacgtcatt
     1141 ggacaaacat gctcaatggc tcaaataatg ttgagaattc ttgggtgtgc ttctctggaa
     1201 acacgtctct tgccgatgat ccaaatgttc tctccaattt ctctgcgttg gcacaacaaa
     1261 gagttaacca ttttgataca attgtgcagg aaagggacaa taggaatgcg agtttcctga
     1321 atggttcagc agttggaaat aacctgaaca cttcgtttag cgtttttggc aacttgcgag
     1381 gaggagacca agatcatcga gttctttccg atcacactgg aatttacact ggtttgggat
     1441 ccactggaca aaacgctgtt ggaaccggga taagactggc cgctgatgtc gcgaacggca
     1501 atttcttgga gcagagagca cctactgcca tgggacacaa ccaaagctat tctgcactga
     1561 accagtcatt agcaccaaca ctccttgacc aatacaatca ggcactcctc gaccaataca
     1621 atcagtcttc acaaatggct caaggacgag gatatccggc tccaaatata gcttacggtc
     1681 ttcaaaatgc cggttttcca gctccacaaa tagctcatcg gccgaatact caaaatgcag
     1741 atcctcaagc gatgaatatg aataaccgtt taagagatca taccttccag atgccacata
     1801 caaacgctca agttccaatg agttctctgc caggtctatt taatttatcc atgaatcacg
     1861 gctccgggaa tcaacagttc caaatgaatc aaagctcatc aggccctgca cctcagttgc
     1921 ctaatctgtc ggaaagcttc cagatggcgc aagggagctc tcaagttaca atgagttcca
     1981 gaccagctca ctccaacact tctatgaatc acagctcgag gaaccaatac aaccacgtgg
     2041 atcaacgccc atcacgccct gcaccccatt tgccaactct gacaaaggaa gaggaagctt
     2101 tcttggagac tccagatttt gaacaatttg gaagacaatt gtacaactac ttgcttccgg
     2161 gaatccttcc aaatggtgct gcagacactt tcagtgacgc tcccaagcat cagttgctga
     2221 agctggcaaa aacactgaag ccgatgctct acgactactg gcggaagacg atgcaaatgc
     2281 agaatcgtgg agccaacatc aacattattc agtggattat tgatttcaac gccaagtttg
     2341 ctgctctcaa cagttcagtt gccaaagcct caaactcatc ggatgttctc aatcaaactc
     2401 ttccaactgc ggctgaggtt tcggatgttg ccagagagga tgcttcgact tcacagccat
     2461 ccaaaagccg atccatgtac attcgtcccg ctgccagtct tgataatacc ttggagacac
     2521 ttgatgagaa tttggattct tcacaaagtc atgccgggcc agttccagca gcttcaacta
     2581 agcccaaaac accatccttt gagaagatga ttagatacag tggcatcaaa aagcgaagca
     2641 caatggacat ggataatttt gttcgtatgc ttgatgaaaa gatcaacttt tcgcctgagc
     2701 caagcacttc aagtgatatt gccagctctg tgaaaggcta catgtctcaa tccttcctcc
     2761 accaacaaga tgacgaagct cctgattgca ccaagaatgt tcattctgaa agtgatttga
     2821 aacaagctga gccacaagaa tcggacaagc aaagtgacaa ggaattgcca agcaatgaat
     2881 gagcggccgc atcgtgactg actgacgatc tgcctcgcgc gtttcggtga tgacggtgaa
     2941 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     3001 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
     3061 acccagtcac gtagcgatag cggagtgtat aattcttgaa gacgaaaggg cctcgtgata
     3121 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     3181 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3241 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3301 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     3361 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     3421 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3481 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3541 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3601 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3661 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3721 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3781 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     3841 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     3901 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     3961 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4021 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4081 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4141 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4201 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4261 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4321 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4381 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4441 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     4501 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4561 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4621 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4681 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4741 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4801 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4861 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4921 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4981 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     5041 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     5101 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataaat
     5161 tccgacacca tcgaatggtg caaaaccttt cgcggtatgg catgatagcg cccggaagag
     5221 agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc
     5281 ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa
     5341 acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca
     5401 caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg
     5461 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc
     5521 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat
     5581 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc
     5641 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag
     5701 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
     5761 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
     5821 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
     5881 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
     5941 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
     6001 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
     6061 gacgataccg aagacagctc atgttatatc ccgccgtcaa ccaccatcaa acaggatttt
     6121 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
     6181 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat
     6241 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     6301 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     6361 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     6421 ataacaattt cacacaggaa acagctatga ccatgattac ggattcactg gccgtcgttt
     6481 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     6541 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     6601 tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa gcggtgccgg
     6661 aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc tcaaactggc
     6721 agatgcacgg ttacgatgcg cccatctaca ccaacgtaac ctatcccatt acggtcaatc
     6781 cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt aatgttgatg
     6841 aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttgga att
//
