LOCUS       sequence-104083-        7481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of GST-MEG-4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7481)
  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 7481)
  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: pJW4.34
FEATURES             Location/Qualifiers
     source          1..7481
                     /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(3531..3553)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3698..3716)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3784..3888
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3889..3957,3958..4749)
                     /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(4107..4126)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4920..5508
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5409..5428
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5662..5679
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5752..5829
                     /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             5830..6912
                     /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(5849..5868)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    6925..6946
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6961..6966,6967..6984,6985..6991)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6999..7015
                     /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     7023..7039
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7023..7039
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(7035..7481,1..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(7055..7072)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7055..7071)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7064..7086)
                     /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 catgagttct tcaaagcttt cctcatgtga tctgccaaac tccagaggaa
     1021 acagagaagg tcaccgctct catagtcgaa gcaatttgga aagtgcatcg aaaaatatgg
     1081 gacaccacat tagtattgaa gaattgtttt attcgttcgt aaagcaagac ggtgttcagt
     1141 caacaagccg tggatatggt ggcagatctc accacggatc cgtgtctttt aacgggaatc
     1201 cttccatgaa cggcgaccag ttgaatcact cgattaatca atcgatgaat gcagctcgag
     1261 ctaacagaaa cgtcttcaac tcatcgagca ttcctccaac ggcccccgtg ttctctgcag
     1321 acttccgtca aaactcgcag acacgcaata actccagctg gaatcaacgg aacttgtcag
     1381 tttccactta tcagtacaac actcagcagt caaatacagg gcacagtaga agccgtccga
     1441 atggacaact tggccgaagc ttctcggatg gatcgaacaa tcatggccat gctgggaatg
     1501 tgtcattcag tgggagttcc gttccagctg ggttccagaa acaagaaacc aaggcttacc
     1561 aacaacttca tggaaataaa agcttcaacg ctcaagctgg agttcaagct cctggagatc
     1621 acaaggacaa gagatatgga ggcatgtttg gaaatgagct tatcaattcg atcaattcta
     1681 gttttggcca acagcatgat gtcttcaagc cgcggaattt gaaccagagc ttttccggat
     1741 ttactattaa tgaagcttta acaaggaagc atcatcaaga aggatcaaga aacgatagaa
     1801 gaagatcaca tgggagcact aatgtccagg atcgtgcttt ctacaacaca gatgacaatg
     1861 aactgtctga cgacgtgttc atagaagatt ccaatggtgt tgcaaacggt ccaaaatctg
     1921 tcactaacaa atctcaacaa cagtctcgcg gaacgaagga tgctggcagt caagtggcgg
     1981 agaaactgaa ctcttccaga ggagctgctc aacgtaatct gaaatccaat gtgttcagca
     2041 aggacggcac cgaactttct gtggaacagt tactggaaat tgtcagcatg aagatcggac
     2101 aacacttgca ctcagcttcc agttctcacg gagaatgcag caatttgaat cggacattac
     2161 cagctagaag tgatctaaat tgctcaattg gtgaagactt cgactccagt tttctggatg
     2221 taaacaatcg aaccctgcca gaaaatcttt cgaagaaaac ttcagtgcag aaacatcgtc
     2281 gtggaagttc tcgtagtgcg tcccgtctcg ccagtcttga cgtgacgctg gagactgtag
     2341 aagaagatgt cgaacccaca ccaagtccac atccgagttc tcctccaaag atctcaagaa
     2401 gaaagtggac aggaactttt gatgccaacg ttgaggaaat gagacgttta cttcatgggg
     2461 accctgggaa gccgaaatct acaaaacgtg ctttctcaag caaggatcag agaacacctt
     2521 ccttgtcaat catcacaagc ccgaaggctt tgatcggaga aaaatgtctg acagcatcca
     2581 acaagtcttc aaagattgac aagtcacttg gcatgattga ttcgaaactc acaaagtcac
     2641 caatgtactc cgccacggtt tctgggaagg aagcaacttc aggaaaccga atcgctcgaa
     2701 agttgactcc agggcttgat ggactgaaaa gctctcattg taccggaaat ccagtttcca
     2761 tagattgtaa tggttgccca actccgaaaa gaagtggaat caattgtgat aatcgtatgg
     2821 ctgaagtgaa caaccaagcc gggagatggt cctctgaagc caccagtcag cctgccccac
     2881 tcccttgcaa gtcagccaac ccgaccgaag attcttctca agatgcacct ccaacttcac
     2941 caccacctcg tatttctgat agcttgacgg cattcctgga agctcaacaa gacttcaatg
     3001 attatattga tgctcactac aaagagaaga cacaactctt gaaagtgaat ttgaatattc
     3061 acgggatgtc acctgaaaga tggctgtact tgaattattt ttgcaccgag acaattccaa
     3121 gattggatgg tccttatgcc gatgatccaa gggttccacc tgtgcgaaat atattcagaa
     3181 gatggtttct ccggtttgct gaagcctgct taggaaatcc aaatgacctt gcagttcaga
     3241 aagatattgc tctcgagttc gttcaggctc ggcttgacga tacgagcagc tccacagatt
     3301 ccaccaatat gctctacatg ttgtggaagg agtgtatcgg acagaagaat atcatctcaa
     3361 ttgctgatgc gtgtctcctt gtatatcttc ggaaatcgga tcctgtcaga tacttgaatg
     3421 caaaacgaga atggctggaa tctatttttg acccacccag agatcaatga gcggccgcat
     3481 cgtgactgac tgacgatctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     3541 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     3601 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     3661 agcgatagcg gagtgtataa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     3721 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     3781 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3841 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     3901 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3961 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4021 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4081 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     4141 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4201 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4261 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4321 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4381 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     4441 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4501 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4561 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4621 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4681 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4741 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     4801 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4861 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4921 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4981 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5041 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     5101 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5161 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5221 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5281 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5341 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5401 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5461 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5521 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5581 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     5641 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     5701 cggtattttc tccttacgca tctgtgcggt atttcacacc gcataaattc cgacaccatc
     5761 gaatggtgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg
     5821 gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat
     5881 cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa
     5941 gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg
     6001 ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg
     6061 caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg
     6121 atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa
     6181 cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa
     6241 gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac
     6301 agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg
     6361 ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt
     6421 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa
     6481 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc
     6541 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
     6601 accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa
     6661 gacagctcat gttatatccc gccgtcaacc accatcaaac aggattttcg cctgctgggg
     6721 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
     6781 ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc
     6841 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     6901 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     6961 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     7021 cacaggaaac agctatgacc atgattacgg attcactggc cgtcgtttta caacgtcgtg
     7081 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     7141 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     7201 atggcgaatg gcgctttgcc tggtttccgg caccagaagc ggtgccggaa agctggctgg
     7261 agtgcgatct tcctgaggcc gatactgtcg tcgtcccctc aaactggcag atgcacggtt
     7321 acgatgcgcc catctacacc aacgtaacct atcccattac ggtcaatccg ccgtttgttc
     7381 ccacggagaa tccgacgggt tgttactcgc tcacatttaa tgttgatgaa agctggctac
     7441 aggaaggcca gacgcgaatt atttttgatg gcgttggaat t
//
