LOCUS       sequence-263995-        7571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of GST:MEG-3 in C. elegans.
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(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             736..759
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3439..3461)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3606..3624)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3692..3796
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3797..3865,3866..4657)
                     /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(4015..4034)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4828..5416
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5317..5336
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5570..5587
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5660..5737
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5738..6820
                     /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(5757..5776)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6869..6874,6875..6892,6893..6899)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6907..6923
                     /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     6931..6947
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6931..6947
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             6943..7470
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6963..6980)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6963..6979)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6972..6994)
                     /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 cctccaaaat cggatctgga agttctgttc caggggcccc tgggatcccc ggaattcccg
      781 ggtcgactca tgagttcctc aaaaccttac ccaagcggtc tgccaaactc gagaagaaaa
      841 agaggaggtc gccgctcaag tagtcgtagc aatcaggaaa gtgcatccaa caacatggaa
      901 caccaaatca ctcttgacga attgttcaat ccgatcgcaa aacaagacag tgctcagtca
      961 acaagccgtg aatatggtgc aaaatcgggg atatctcatc acggatccgt atcattcaac
     1021 gggaatacat tcatgaatgg ccaacagctg aatcactcga tgactcgaca tggtcgagta
     1081 ttcaatcagt cgatgcatgc agctcaagga aacggcagca acgcctttaa cagtattcct
     1141 cctacggccc ccgtcttctc tgcagacttc cgccgcaact tgcaaacacg caatagctcc
     1201 agctggtatg aacgaagatt cccagtatcc acagatcagg acgacgttca gcaatcaaac
     1261 acaagacgta gtagaagtcg tcagaatgga caacatggcc taagcttctc ggatggttcg
     1321 aacaattatg gtcacgctgg gaataagtca ttcagtgtga gttccgttcc cgttgggttc
     1381 cagaagcaag aaaataattc taagaaactg cgacaaacca atgttcacca gcaatgtctt
     1441 ggaaataaaa gcttcaacgc tcaagctgga gttcatggac acgcttttaa aaaaggccat
     1501 aaggacaata aaaatgcatc tggaaaggag gttatcaatt ctagtcttgt ccaaaagcac
     1561 gatgctatta agtcgcggaa tttgaaccag agcttctctg gatttcctac acatgaaacc
     1621 tcatcgatga aaaatcaaca acagaaatca agaaacgaca gaaaaaaatc acgtggtagc
     1681 agcaacttcc aggatcgtac ttacttcaac acaaatgacg atgaattaac tgacgatgtg
     1741 ttcatagacg attccatgga tgctgcccgt ggtcggagat ctcgatcagt cactaagaaa
     1801 cttcaacaat cgacttattc aaagcagaat gcaggcagta aacaattgac tgaaaaatgc
     1861 aaatcttctg aagaagcggc taaacgtaac ctggtatcca atgtattcag caaggacggc
     1921 actgaactct ccattgaaca gttattggaa attgtcagca tgaagatcgg acaacagatt
     1981 cacttgcctt ccagctctca tggagaatgc agcaatttga atcggacatt accagcaagt
     2041 gatctaaatt gctcaattgg tgaagacttt gactccagtt ttgtggatgc taacaaccaa
     2101 accctaccag tgagtcttcc caagaaaact tcactgtcga tcaagcgccg tggaagttct
     2161 cgtagtgcct cccgtctggc cagtcttgat gtgacactgg aaactgttga agaagacgag
     2221 gaacccacgc caagtccaca gccgagttct cctccaaaga tctcaagaag aaagtggaca
     2281 ggaacttttg atgccaacgt tgaggaaatg agacgtttac ttcatgggga ccctgagatg
     2341 ccgaaatctg caaatcgtgc ttcctcaagc aaggatcaga tcaatcggaa caatgtggat
     2401 gtaaagagaa caccttcctc gtcaatcatc ccaactccaa aggctttgat cggagaaaga
     2461 tgtctgacat catccagcaa gtcttcaaag ttgaacaagt cacttggcgt ggttgattct
     2521 aaagctacaa aatcaccaat gtattccgtc acagtttctg gaaaagaaac agcttcagga
     2581 aagcgaattg ctcaaaagtt gactccaaag gttgtggcgc tggaaagctc ttacattacc
     2641 ggaattccag tttccacgga ttgtaatggt tgcccaactc cgaagagaag tggaatcaat
     2701 tgtgaaattc gtgcagctga agtatacaat caagctggga aatggccctt cgaaatcacc
     2761 agtgatccgg ctccacttcc ttgtgaatca gccgatcgga tcgaatatcc ttctcaagat
     2821 tgcactcaag atccagcttc aacctcgcca ccacctcgca tttctgaaag cttgacagca
     2881 ttcctggaag ctcagcaaga ctttaacgac tacattgata ctaactacaa agaaaagaca
     2941 caactcctga aagtgaattt gaatattcac gggatgtcac ctgaaagatg gctgtacttg
     3001 aattattttt gcaccgagac aattccaaga ttggatggtc cttatgccga tgatccaaga
     3061 gttccacctg tgcgaaatat gttcagaaaa tggttcctcc gttttgctga agcctgctta
     3121 ggaaatcctc accagcttgc agttatgcaa gagattgctg ccacgtttgt tcaggctcgg
     3181 ctggacgata cgagcagctc cacagactcc accaatatgc tctacatgtt gtggaaggag
     3241 tgtatcggac agaagaatat catcgcaatt gctgatgcgt gtttacttgc acatcttcgg
     3301 aaatcggatc ctatcaaata cttgaatgta aaacgggatt ggctggaatc tatttttgac
     3361 ccacccagag atcaatgagc ggccgcatcg tgactgactg acgatctgcc tcgcgcgttt
     3421 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     3481 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     3541 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtataatt cttgaagacg
     3601 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     3661 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3721 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3781 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3841 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3901 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3961 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4021 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4081 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4141 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4201 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4261 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4321 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     4381 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4441 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4501 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4561 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4621 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4681 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4741 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4801 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4861 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4921 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     4981 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5041 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5101 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5161 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5221 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5281 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5341 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5401 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5461 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5521 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     5581 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     5641 ttcacaccgc ataaattccg acaccatcga atggcgcaaa acctttcgcg gtatggcatg
     5701 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
     5761 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
     5821 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     5881 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     5941 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     6001 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     6061 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     6121 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     6181 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     6241 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     6301 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     6361 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     6421 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     6481 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
     6541 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac
     6601 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     6661 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
     6721 cacgctagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     6781 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa
     6841 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     6901 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacggat
     6961 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     7021 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     7081 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gctttgcctg gtttccggca
     7141 ccagaagcgg tgccggaaag ctggctggag tgcgatcttc ctgaggccga tactgtcgtc
     7201 gtcccctcaa actggcagat gcacggttac gatgcgccca tctacaccaa cgtgacctat
     7261 cccattacgg tcaatccgcc gtttgttccc acggagaatc cgacgggttg ttactcgctc
     7321 acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat ttttgatggc
     7381 gttggaatta gcttatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg
     7441 gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg
     7501 cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct
     7561 gaaatgagct g
//
