LOCUS       sequence-260060-        8333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of MBP:PPTR-1 in C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8333)
  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 8333)
  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: pJW4.12
FEATURES             Location/Qualifiers
     source          1..8333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    775..917
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(843..862)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1003..1025
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1019..1210)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        1706..1783
                     /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             1784..2866
                     /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(1803..1822)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(3109..3114,3115..3130,3131..3137)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3145..3161
                     /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     3150..3172
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             3231..4331
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     4305..4328
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             4380..4391
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    4395..4419
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6052..6076)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(6127..6144)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6127..6143)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6136..6158)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6322..6339)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      6492..6578
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6670..6697
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        6716..6807
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6808..6876,6877..7668)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7026..7045)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7710..8223)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7905..7926)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8117..8136
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatatg gtgcactctc
      841 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
      901 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
      961 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     1021 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     1081 ggtcgtgcag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     1141 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     1201 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga
     1261 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg
     1321 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc
     1381 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc
     1441 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac
     1501 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc
     1561 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc
     1621 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtggcc
     1681 aggacccaac gctgcccgaa attccgacac catcgaatgg tgcaaaacct ttcgcggtat
     1741 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
     1801 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
     1861 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
     1921 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
     1981 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
     2041 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
     2101 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
     2161 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
     2221 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
     2281 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
     2341 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
     2401 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
     2461 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
     2521 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
     2581 ggatatctca gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
     2641 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
     2701 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
     2761 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     2821 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     2881 atgtaagtta gctcactcat taggcacaat tctcatgttt gacagcttat catcgactgc
     2941 acggtgcacc aatgcttctg gcgtcaggca gccatcggaa gctgtggtat ggctgtgcag
     3001 gtcgtaaatc actgcataat tcgtgtcgct caaggcgcac tcccgttctg gataatgttt
     3061 tttgcgccga catcataacg gttctggcaa atattctgaa atgagctgtt gacaattaat
     3121 catcggctcg tataatgtgt ggaattgtga gcggataaca atttcacaca ggaaacagcc
     3181 agtccgttta ggtgttttca cgagcacttc accaacaagg accatagcat atgaaaatcg
     3241 aagaaggtaa actggtaatc tggattaacg gcgataaagg ctataacggt ctcgctgaag
     3301 tcggtaagaa attcgagaaa gataccggaa ttaaagtcac cgttgagcat ccggataaac
     3361 tggaagagaa attcccacag gttgcggcaa ctggcgatgg ccctgacatt atcttctggg
     3421 cacacgaccg ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc accccggaca
     3481 aagcgttcca ggacaagctg tatccgttta cctgggatgc cgtacgttac aacggcaagc
     3541 tgattgctta cccgatcgct gttgaagcgt tatcgctgat ttataacaaa gatctgctgc
     3601 cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga taaagaactg aaagcgaaag
     3661 gtaagagcgc gctgatgttc aacctgcaag aaccgtactt cacctggccg ctgattgctg
     3721 ctgacggggg ttatgcgttc aagtatgaaa acggcaagta cgacattaaa gacgtgggcg
     3781 tggataacgc tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt aaaaacaaac
     3841 acatgaatgc agacaccgat tactccatcg cagaagctgc ctttaataaa ggcgaaacag
     3901 cgatgaccat caacggcccg tgggcatggt ccaacatcga caccagcaaa gtgaattatg
     3961 gtgtaacggt actgccgacc ttcaagggtc aaccatccaa accgttcgtt ggcgtgctga
     4021 gcgcaggtat taacgccgcc agtccgaaca aagagctggc aaaagagttc ctcgaaaact
     4081 atctgctgac tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg ggtgccgtag
     4141 cgctgaagtc ttacgaggaa gagttggcga aagatccacg tattgccgcc actatggaaa
     4201 acgcccagaa aggtgaaatc atgccgaaca tcccgcagat gtccgctttc tggtatgccg
     4261 tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa gccctgaaag
     4321 acgcgcagac taattcgagc tcgaacaaca acaacaataa caataacaac aacctcggga
     4381 tcgagggaag gatcacaagt ttgtacaaaa aagcaggctt aatgcacgga agcgggcaca
     4441 gtctaacggg cgctccgcat caaattccgc cgccacggac acagggagcc gccaccggtg
     4501 gccaacaact ctcggcgaca gctaatcaat ttgttgataa aatcgatccg ttccataata
     4561 aaagaggaac ttctagacga ttgcgcatca acaacagcag ccgatataat gttgattcgg
     4621 ctcaagaact cgtacaattg gcgctcatca aagatactgc tgcaaatgag caaccggctc
     4681 tcgttatcga aaagctcgtc caatgtcagc acgtattcga tttctatgat ccagtagctc
     4741 aactgaaatg caaagaaatc aaaagagcag ctctaaatga gctcatcgat cacattacat
     4801 cgacaaaagg agcaattgtt gagacaattt atccggctgt cattaaaatg gttgccaaga
     4861 atatattccg tgtacttcca ccatcagaaa attgtgaatt tgatccagaa gaggatgaac
     4921 cgactttaga agtctcgtgg ccacatttac agcttgtcta cgagctcttc ctgagattcc
     4981 ttgagtcgcc agatttccaa gcttcaatcg gtaaaaagta tattgatcag agatttgtgc
     5041 tcaagctgct cgatttattc gattcggagg atccacgtga acgagacttt ttgaagacgg
     5101 ttcttcatcg tatttacgga aaatttttgg gccttcgggc ttttattcgc aaacatatca
     5161 ataatatgtt tttgagattc gtatacgaaa cggactcatt caacggtgtc ggtgagcttc
     5221 tcgagattct tggctcaatt ataaacggat ttgctcttcc attgaagcaa gagcacaagg
     5281 ttttccttgt aaaagttctt ctaccattgc acaaaccgaa atgcttatca ctgtatcatg
     5341 cacagctcgc ctactgcgtc gttcaattca tcgaaaaaga ctcatcactg actccacaag
     5401 tttttgaggc tctgctcaaa ttttggcctc gaacatgtag cagcaaggag gttatgttcc
     5461 tgggtgaagt ggaggaaatt ctcgacatta tcgaaccgga acaattcaaa aagattatcg
     5521 atccattatt ccgtcaattg gccaaatgtg tcagtagtcc acattttcaa gttgctgaac
     5581 gggctctcta cttttggaat aatgaatata tattgtcatt aattgaggat acaagcagtt
     5641 tggtgatgcc gattatgttc ccagcgctct atcggatttc caaagagcac tggaatcaaa
     5701 caattgttgc acttgtctat aatgtactca aaacttttat ggaaatgaat ggaaaactgt
     5761 ttgatgagct tacttctacg tacaaaggtg aacgattgcg ggaaaagcaa cgagaaaagg
     5821 atcgtgatgc tttttggaag aaaatggaag ctcttgaatt gaatccaccg gctgaaggaa
     5881 aagaagtgac accttcgttg tttccagaga agttgactga ttatttgaag aaggacggtc
     5941 caaatatgac gccactgcca gttgcaactg caggtggcgg tgataaatct ccatctgtag
     6001 tgaagaaatc gtcaacagga tcagaaacga cgacgccggc taaaaaataa gacccagctt
     6061 tcttgtacaa agtggtgata tttcagaatt cggatcctct agagtcgacc tgcaggcaag
     6121 cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     6181 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     6241 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcagcttg gctgttttgg
     6301 cggatgagat aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat
     6361 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     6421 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
     6481 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     6541 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     6601 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     6661 aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt
     6721 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6781 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6841 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6901 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6961 ggtaagatcc ttgagagttt tcgccccgaa gaacgtttcc caatgatgag cacttttaaa
     7021 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     7081 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     7141 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     7201 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     7261 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     7321 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     7381 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     7441 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     7501 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     7561 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     7621 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     7681 gtttactcat atatacttta gattgattta ccccggttga taatcagaaa agccccaaaa
     7741 acaggaagat tgtataagca aatatttaaa ttgtaaacgt taatattttg ttaaaattcg
     7801 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     7861 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     7921 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     7981 atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag
     8041 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     8101 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     8161 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     8221 cgtaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     8281 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttc
//
