LOCUS       sequence-104084-        7571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of GST-MEG-3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7571)
  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 7571)
  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: pJW4.33
FEATURES             Location/Qualifiers
     source          1..7571
                     /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(3621..3643)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3788..3806)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3874..3978
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3979..4047,4048..4839)
                     /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(4197..4216)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5010..5598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5499..5518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5752..5769
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5842..5919
                     /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             5920..7002
                     /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(5939..5958)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    7015..7036
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7051..7056,7057..7074,7075..7081)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7089..7105
                     /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     7113..7129
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7113..7129
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(7125..7571,1..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(7145..7162)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7145..7161)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7154..7176)
                     /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 catgagttcc tcaaaacctt acccaagcgg tctgccaaac tcgagaagaa
     1021 aaagaggagg tcgccgctca agtagtcgta gcaatcagga aagtgcatcc aacaacatgg
     1081 aacaccaaat cactcttgac gaattgttca atccgatcgc aaaacaagac agtgctcagt
     1141 caacaagccg tgaatatggt gcaaaatcgg ggatatctca tcacggatcc gtatcattca
     1201 acgggaatac attcatgaat ggccaacagc tgaatcactc gatgactcga catggtcgag
     1261 tattcaatca gtcgatgcat gcagctcaag gaaacggcag caacgccttt aacagtattc
     1321 ctcctacggc ccccgtcttc tctgcagact tccgccgcaa cttgcaaaca cgcaatagct
     1381 ccagctggta tgaacgaaga ttcccagtat ccacagatca ggacgacgtt cagcaatcaa
     1441 acacaagacg tagtagaagt cgtcagaatg gacaacatgg cctaagcttc tcggatggtt
     1501 cgaacaatta tggtcacgct gggaataagt cattcagtgt gagttccgtt cccgttgggt
     1561 tccagaagca agaaaataat tctaagaaac tgcgacaaac caatgttcac cagcaatgtc
     1621 ttggaaataa aagcttcaac gctcaagctg gagttcatgg acacgctttt aaaaaaggcc
     1681 ataaggacaa taaaaatgca tctggaaagg aggttatcaa ttctagtctt gtccaaaagc
     1741 acgatgctat taagtcgcgg aatttgaacc agagcttctc tggatttcct acacatgaaa
     1801 cctcatcgat gaaaaatcaa caacagaaat caagaaacga cagaaaaaaa tcacgtggta
     1861 gcagcaactt ccaggatcgt acttacttca acacaaatga cgatgaatta actgacgatg
     1921 tgttcataga cgattccatg gatgctgccc gtggtcggag atctcgatca gtcactaaga
     1981 aacttcaaca atcgacttat tcaaagcaga atgcaggcag taaacaattg actgaaaaat
     2041 gcaaatcttc tgaagaagcg gctaaacgta acctggtatc caatgtattc agcaaggacg
     2101 gcactgaact ctccattgaa cagttattgg aaattgtcag catgaagatc ggacaacaga
     2161 ttcacttgcc ttccagctct catggagaat gcagcaattt gaatcggaca ttaccagcaa
     2221 gtgatctaaa ttgctcaatt ggtgaagact ttgactccag ttttgtggat gctaacaacc
     2281 aaaccctacc agtgagtctt cccaagaaaa cttcactgtc gatcaagcgc cgtggaagtt
     2341 ctcgtagtgc ctcccgtctg gccagtcttg atgtgacact ggaaactgtt gaagaagacg
     2401 aggaacccac gccaagtcca cagccgagtt ctcctccaaa gatctcaaga agaaagtgga
     2461 caggaacttt tgatgccaac gttgaggaaa tgagacgttt acttcatggg gaccctgaga
     2521 tgccgaaatc tgcaaatcgt gcttcctcaa gcaaggatca gatcaatcgg aacaatgtgg
     2581 atgtaaagag aacaccttcc tcgtcaatca tcccaactcc aaaggctttg atcggagaaa
     2641 gatgtctgac atcatccagc aagtcttcaa agttgaacaa gtcacttggc gtggttgatt
     2701 ctaaagctac aaaatcacca atgtattccg tcacagtttc tggaaaagaa acagcttcag
     2761 gaaagcgaat tgctcaaaag ttgactccaa aggttgtggc gctggaaagc tcttacatta
     2821 ccggaattcc agtttccacg gattgtaatg gttgcccaac tccgaagaga agtggaatca
     2881 attgtgaaat tcgtgcagct gaagtataca atcaagctgg gaaatggccc ttcgaaatca
     2941 ccagtgatcc ggctccactt ccttgtgaat cagccgatcg gatcgaatat ccttctcaag
     3001 attgcactca agatccagct tcaacctcgc caccacctcg catttctgaa agcttgacag
     3061 cattcctgga agctcagcaa gactttaacg actacattga tactaactac aaagaaaaga
     3121 cacaactcct gaaagtgaat ttgaatattc acgggatgtc acctgaaaga tggctgtact
     3181 tgaattattt ttgcaccgag acaattccaa gattggatgg tccttatgcc gatgatccaa
     3241 gagttccacc tgtgcgaaat atgttcagaa aatggttcct ccgttttgct gaagcctgct
     3301 taggaaatcc tcaccagctt gcagttatgc aagagattgc tgccacgttt gttcaggctc
     3361 ggctggacga tacgagcagc tccacagact ccaccaatat gctctacatg ttgtggaagg
     3421 agtgtatcgg acagaagaat atcatcgcaa ttgctgatgc gtgtttactt gcacatcttc
     3481 ggaaatcgga tcctatcaaa tacttgaatg taaaacggga ttggctggaa tctatttttg
     3541 acccacccag agatcaatga gcggccgcat cgtgactgac tgacgatctg cctcgcgcgt
     3601 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     3661 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     3721 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtataa ttcttgaaga
     3781 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     3841 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3901 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3961 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4021 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4081 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4141 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4201 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     4261 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4321 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4381 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4441 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4501 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     4561 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4621 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4681 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4741 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4801 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4861 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4921 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4981 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5041 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5101 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     5161 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5221 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5281 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5341 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5401 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5461 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5521 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5581 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5641 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5701 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     5761 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     5821 atttcacacc gcataaattc cgacaccatc gaatggtgca aaacctttcg cggtatggca
     5881 tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac
     5941 gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc
     6001 agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac
     6061 attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc
     6121 acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc
     6181 gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt
     6241 aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg
     6301 ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt
     6361 cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg
     6421 cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc
     6481 ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc
     6541 aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa
     6601 caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat
     6661 cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat
     6721 atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc gccgtcaacc
     6781 accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc
     6841 tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa
     6901 accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     6961 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     7021 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     7081 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgg
     7141 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     7201 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     7261 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgctttgcc tggtttccgg
     7321 caccagaagc ggtgccggaa agctggctgg agtgcgatct tcctgaggcc gatactgtcg
     7381 tcgtcccctc aaactggcag atgcacggtt acgatgcgcc catctacacc aacgtaacct
     7441 atcccattac ggtcaatccg ccgtttgttc ccacggagaa tccgacgggt tgttactcgc
     7501 tcacatttaa tgttgatgaa agctggctac aggaaggcca gacgcgaatt atttttgatg
     7561 gcgttggaat t
//
