LOCUS       sequence-104099-        8333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of MBP-PPTR-1.
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"
     promoter        3..80
                     /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             81..1163
                     /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(100..119)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1406..1411,1412..1427,1428..1434)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1442..1458
                     /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     1447..1469
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1528..2628
                     /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     2602..2625
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2677..2688
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    2692..2716
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4349..4373)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4424..4441)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4424..4440)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4433..4455)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4619..4636)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4789..4875
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4967..4994
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5013..5104
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5105..5173,5174..5965)
                     /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(5323..5342)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6007..6520)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6202..6223)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6414..6433
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      6631..7219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7120..7139
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7373..7390
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7405..7547
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7473..7492)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7633..7655
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7649..7840)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
       61 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
      121 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
      181 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
      241 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
      301 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
      361 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
      421 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
      481 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
      541 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
      601 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
      661 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
      721 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
      781 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
      841 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
      901 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
      961 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     1021 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     1081 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     1141 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     1201 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg
     1261 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     1321 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     1381 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
     1441 attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga
     1501 gcacttcacc aacaaggacc atagcatatg aaaatcgaag aaggtaaact ggtaatctgg
     1561 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat
     1621 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt
     1681 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac
     1741 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat
     1801 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt
     1861 gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa
     1921 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac
     1981 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag
     2041 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg
     2101 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac
     2161 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg
     2221 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc
     2281 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt
     2341 ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg
     2401 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag
     2461 ttggcgaaag atccacgtat tgccgccact atggaaaacg cccagaaagg tgaaatcatg
     2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc
     2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg
     2641 aacaacaaca acaataacaa taacaacaac ctcgggatcg agggaaggat cacaagtttg
     2701 tacaaaaaag caggcttaat gcacggaagc gggcacagtc taacgggcgc tccgcatcaa
     2761 attccgccgc cacggacaca gggagccgcc accggtggcc aacaactctc ggcgacagct
     2821 aatcaatttg ttgataaaat cgatccgttc cataataaaa gaggaacttc tagacgattg
     2881 cgcatcaaca acagcagccg atataatgtt gattcggctc aagaactcgt acaattggcg
     2941 ctcatcaaag atactgctgc aaatgagcaa ccggctctcg ttatcgaaaa gctcgtccaa
     3001 tgtcagcacg tattcgattt ctatgatcca gtagctcaac tgaaatgcaa agaaatcaaa
     3061 agagcagctc taaatgagct catcgatcac attacatcga caaaaggagc aattgttgag
     3121 acaatttatc cggctgtcat taaaatggtt gccaagaata tattccgtgt acttccacca
     3181 tcagaaaatt gtgaatttga tccagaagag gatgaaccga ctttagaagt ctcgtggcca
     3241 catttacagc ttgtctacga gctcttcctg agattccttg agtcgccaga tttccaagct
     3301 tcaatcggta aaaagtatat tgatcagaga tttgtgctca agctgctcga tttattcgat
     3361 tcggaggatc cacgtgaacg agactttttg aagacggttc ttcatcgtat ttacggaaaa
     3421 tttttgggcc ttcgggcttt tattcgcaaa catatcaata atatgttttt gagattcgta
     3481 tacgaaacgg actcattcaa cggtgtcggt gagcttctcg agattcttgg ctcaattata
     3541 aacggatttg ctcttccatt gaagcaagag cacaaggttt tccttgtaaa agttcttcta
     3601 ccattgcaca aaccgaaatg cttatcactg tatcatgcac agctcgccta ctgcgtcgtt
     3661 caattcatcg aaaaagactc atcactgact ccacaagttt ttgaggctct gctcaaattt
     3721 tggcctcgaa catgtagcag caaggaggtt atgttcctgg gtgaagtgga ggaaattctc
     3781 gacattatcg aaccggaaca attcaaaaag attatcgatc cattattccg tcaattggcc
     3841 aaatgtgtca gtagtccaca ttttcaagtt gctgaacggg ctctctactt ttggaataat
     3901 gaatatatat tgtcattaat tgaggataca agcagtttgg tgatgccgat tatgttccca
     3961 gcgctctatc ggatttccaa agagcactgg aatcaaacaa ttgttgcact tgtctataat
     4021 gtactcaaaa cttttatgga aatgaatgga aaactgtttg atgagcttac ttctacgtac
     4081 aaaggtgaac gattgcggga aaagcaacga gaaaaggatc gtgatgcttt ttggaagaaa
     4141 atggaagctc ttgaattgaa tccaccggct gaaggaaaag aagtgacacc ttcgttgttt
     4201 ccagagaagt tgactgatta tttgaagaag gacggtccaa atatgacgcc actgccagtt
     4261 gcaactgcag gtggcggtga taaatctcca tctgtagtga agaaatcgtc aacaggatca
     4321 gaaacgacga cgccggctaa aaaataagac ccagctttct tgtacaaagt ggtgatattt
     4381 cagaattcgg atcctctaga gtcgacctgc aggcaagctt ggcactggcc gtcgttttac
     4441 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     4501 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     4561 gcagcctgaa tggcgaatgg cagcttggct gttttggcgg atgagataag attttcagcc
     4621 tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca
     4681 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
     4741 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
     4801 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     4861 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
     4921 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
     4981 acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa
     5041 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5101 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5161 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5221 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5281 ccccgaagaa cgtttcccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5341 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5401 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5461 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5521 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5581 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5641 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5701 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5761 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5821 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5881 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5941 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6001 tgatttaccc cggttgataa tcagaaaagc cccaaaaaca ggaagattgt ataagcaaat
     6061 atttaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     6121 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     6181 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     6241 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     6301 cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     6361 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     6421 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     6481 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt aaaaggatct aggtgaagat
     6541 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6601 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6661 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6721 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     6781 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6841 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6901 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6961 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     7021 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     7081 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     7141 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7201 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     7261 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     7321 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     7381 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     7441 tatttcacac cgcatatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     7501 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     7561 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     7621 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     7681 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgcag cgattcacag atgtctgcct
     7741 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     7801 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
     7861 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
     7921 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
     7981 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
     8041 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
     8101 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
     8161 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
     8221 tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg
     8281 acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgaa att
//
