LOCUS       sequence-288112-        7481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of GST:MEG-4 in C. elegans.
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(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(3349..3371)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3516..3534)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3602..3706
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3707..3775,3776..4567)
                     /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(3925..3944)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4738..5326
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5227..5246
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5480..5497
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5570..5647
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5648..6730
                     /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(5667..5686)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6779..6784,6785..6802,6803..6809)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6817..6833
                     /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     6841..6857
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6841..6857
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             6853..7380
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6873..6890)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6873..6889)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6882..6904)
                     /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 tgagttcttc aaagctttcc tcatgtgatc tgccaaactc cagaggaaac
      841 agagaaggtc accgctctca tagtcgaagc aatttggaaa gtgcatcgaa aaatatggga
      901 caccacatta gtattgaaga attgttttat tcgttcgtaa agcaagacgg tgttcagtca
      961 acaagccgtg gatatggtgg cagatctcac cacggatccg tgtcttttaa cgggaatcct
     1021 tccatgaacg gcgaccagtt gaatcactcg attaatcaat cgatgaatgc agctcgagct
     1081 aacagaaacg tcttcaactc atcgagcatt cctccaacgg cccccgtgtt ctctgcagac
     1141 ttccgtcaaa actcgcagac acgcaataac tccagctgga atcaacggaa cttgtcagtt
     1201 tccacttatc agtacaacac tcagcagtca aatacagggc acagtagaag ccgtccgaat
     1261 ggacaacttg gccgaagctt ctcggatgga tcgaacaatc atggccatgc tgggaatgtg
     1321 tcattcagtg ggagttccgt tccagctggg ttccagaaac aagaaaccaa ggcttaccaa
     1381 caacttcatg gaaataaaag cttcaacgct caagctggag ttcaagctcc tggagatcac
     1441 aaggacaaga gatatggagg catgtttgga aatgagctta tcaattcgat caattctagt
     1501 tttggccaac agcatgatgt cttcaagccg cggaatttga accagagctt ttccggattt
     1561 actattaatg aagctttaac aaggaagcat catcaagaag gatcaagaaa cgatagaaga
     1621 agatcacatg ggagcactaa tgtccaggat cgtgctttct acaacacaga tgacaatgaa
     1681 ctgtctgacg acgtgttcat agaagattcc aatggtgttg caaacggtcc aaaatctgtc
     1741 actaacaaat ctcaacaaca gtctcgcgga acgaaggatg ctggcagtca agtggcggag
     1801 aaactgaact cttccagagg agctgctcaa cgtaatctga aatccaatgt gttcagcaag
     1861 gacggcaccg aactttctgt ggaacagtta ctggaaattg tcagcatgaa gatcggacaa
     1921 cacttgcact cagcttccag ttctcacgga gaatgcagca atttgaatcg gacattacca
     1981 gctagaagtg atctaaattg ctcaattggt gaagacttcg actccagttt tctggatgta
     2041 aacaatcgaa ccctgccaga aaatctttcg aagaaaactt cagtgcagaa acatcgtcgt
     2101 ggaagttctc gtagtgcgtc ccgtctcgcc agtcttgacg tgacgctgga gactgtagaa
     2161 gaagatgtcg aacccacacc aagtccacat ccgagttctc ctccaaagat ctcaagaaga
     2221 aagtggacag gaacttttga tgccaacgtt gaggaaatga gacgtttact tcatggggac
     2281 cctgggaagc cgaaatctac aaaacgtgct ttctcaagca aggatcagag aacaccttcc
     2341 ttgtcaatca tcacaagccc gaaggctttg atcggagaaa aatgtctgac agcatccaac
     2401 aagtcttcaa agattgacaa gtcacttggc atgattgatt cgaaactcac aaagtcacca
     2461 atgtactccg ccacggtttc tgggaaggaa gcaacttcag gaaaccgaat cgctcgaaag
     2521 ttgactccag ggcttgatgg actgaaaagc tctcattgta ccggaaatcc agtttccata
     2581 gattgtaatg gttgcccaac tccgaaaaga agtggaatca attgtgataa tcgtatggct
     2641 gaagtgaaca accaagccgg gagatggtcc tctgaagcca ccagtcagcc tgccccactc
     2701 ccttgcaagt cagccaaccc gaccgaagat tcttctcaag atgcacctcc aacttcacca
     2761 ccacctcgta tttctgatag cttgacggca ttcctggaag ctcaacaaga cttcaatgat
     2821 tatattgatg ctcactacaa agagaagaca caactcttga aagtgaattt gaatattcac
     2881 gggatgtcac ctgaaagatg gctgtacttg aattattttt gcaccgagac aattccaaga
     2941 ttggatggtc cttatgccga tgatccaagg gttccacctg tgcgaaatat attcagaaga
     3001 tggtttctcc ggtttgctga agcctgctta ggaaatccaa atgaccttgc agttcagaaa
     3061 gatattgctc tcgagttcgt tcaggctcgg cttgacgata cgagcagctc cacagattcc
     3121 accaatatgc tctacatgtt gtggaaggag tgtatcggac agaagaatat catctcaatt
     3181 gctgatgcgt gtctccttgt atatcttcgg aaatcggatc ctgtcagata cttgaatgca
     3241 aaacgagaat ggctggaatc tatttttgac ccacccagag atcaatgagc ggccgcatcg
     3301 tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     3361 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     3421 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     3481 cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     3541 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     3601 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3661 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3721 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3781 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3841 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3901 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     3961 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4021 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4081 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     4141 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4201 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     4261 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4321 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4381 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4441 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4501 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4561 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     4621 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4681 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4741 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4801 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4861 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     4921 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4981 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5041 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5101 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5161 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5221 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5281 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5341 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5401 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     5461 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     5521 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg acaccatcga
     5581 atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt
     5641 ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca
     5701 gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt
     5761 ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg
     5821 caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca
     5881 aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat
     5941 ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg
     6001 cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc
     6061 tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag
     6121 tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg
     6181 tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct
     6241 ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg
     6301 cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt
     6361 tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac
     6421 cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga
     6481 cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca
     6541 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
     6601 gttgcccgtc tcactggtga aaagaaaaac cacgctagcg cccaatacgc aaaccgcctc
     6661 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     6721 cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca ccccaggctt
     6781 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     6841 caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca acgtcgtgac
     6901 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     6961 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     7021 ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag ctggctggag
     7081 tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat gcacggttac
     7141 gatgcgccca tctacaccaa cgtgacctat cccattacgg tcaatccgcc gtttgttccc
     7201 acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag ctggctacag
     7261 gaaggccaga cgcgaattat ttttgatggc gttggaatta gcttatcgac tgcacggtgc
     7321 accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa
     7381 atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc
     7441 cgacatcata acggttctgg caaatattct gaaatgagct g
//
