LOCUS       sequence-104095-        8132 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of MBP-MEI-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8132)
  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 8132)
  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: pJW4.18
FEATURES             Location/Qualifiers
     source          1..8132
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..1101
                     /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     1075..1098
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1150..1161
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    1165..1189
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2621..2645)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2696..2713)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2696..2712)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2705..2727)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2891..2908)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3061..3147
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3239..3266
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3285..3376
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3377..3445,3446..4237)
                     /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(3595..3614)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4279..4792)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4474..4495)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4686..4705
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4903..5491
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5392..5411
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5645..5662
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5677..5819
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5745..5764)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5905..5927
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5921..6112)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        6608..6685
                     /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             6686..7768
                     /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(6705..6724)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(8011..8016,8017..8032,8033..8039)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    8047..8063
                     /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     8052..8074
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 atgaaaatcg aagaaggtaa actggtaatc tggattaacg gcgataaagg ctataacggt
       61 ctcgctgaag tcggtaagaa attcgagaaa gataccggaa ttaaagtcac cgttgagcat
      121 ccggataaac tggaagagaa attcccacag gttgcggcaa ctggcgatgg ccctgacatt
      181 atcttctggg cacacgaccg ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc
      241 accccggaca aagcgttcca ggacaagctg tatccgttta cctgggatgc cgtacgttac
      301 aacggcaagc tgattgctta cccgatcgct gttgaagcgt tatcgctgat ttataacaaa
      361 gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga taaagaactg
      421 aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag aaccgtactt cacctggccg
      481 ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa acggcaagta cgacattaaa
      541 gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt
      601 aaaaacaaac acatgaatgc agacaccgat tactccatcg cagaagctgc ctttaataaa
      661 ggcgaaacag cgatgaccat caacggcccg tgggcatggt ccaacatcga caccagcaaa
      721 gtgaattatg gtgtaacggt actgccgacc ttcaagggtc aaccatccaa accgttcgtt
      781 ggcgtgctga gcgcaggtat taacgccgcc agtccgaaca aagagctggc aaaagagttc
      841 ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg
      901 ggtgccgtag cgctgaagtc ttacgaggaa gagttggcga aagatccacg tattgccgcc
      961 actatggaaa acgcccagaa aggtgaaatc atgccgaaca tcccgcagat gtccgctttc
     1021 tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa
     1081 gccctgaaag acgcgcagac taattcgagc tcgaacaaca acaacaataa caataacaac
     1141 aacctcggga tcgagggaag gatcacaagt ttgtacaaaa aagcaggctt aatgaatggg
     1201 gatgtgcagt cagtcattcg cggatatctg gagcgagccc aagtcgcgaa aacaatgtcc
     1261 gatgccggac gatggaatga ggctggtgat cttcttcgtc agctcatgac agatgtcaaa
     1321 agctgtaaaa tttcggcaag caacagagac gagcatgacg caagaaatac attcctacga
     1381 gccctagagg ctaatttgaa gcttgtccaa caaaatgtgc gcgatgaaga tgacctacac
     1441 gaggcgatga cgcgacaaag tggaagtcca gagccacctg cagacccaga tgtttggtcc
     1501 aaaccatcgc caccgttacc atcctcatcg aaatttggag caacaaagaa aggagttgga
     1561 gcagcaggtc cacgtcccag agaaatatca aagtcaacgt cgtcgatgag cacgaatccg
     1621 gctgatgtga agcctgcgaa tccaacacaa ggaatactgc ctcaaaatag tgctggagat
     1681 tcattcgatg catcggctta tgatgcgtac attgttcaag cggtgcgtgg tacaatggct
     1741 acaaatacgg aaaatacaat gtctttagat gatataatcg gaatgcacga cgtgaagcaa
     1801 gttctgcacg aagccgtcac tcttccactc ttggtgccag aattcttcca gggtctccgt
     1861 tcaccatgga aagcaatggt tctcgctgga ccacctggaa ctggaaagac tcttatcgca
     1921 cgtgcgattg cttcagagtc tagttcaact ttttttaccg tttcctcgac tgatctgtcc
     1981 agcaagtggc gtggtgattc cgagaagatt gttcgtctgt tattcgaact tgcacggttc
     2041 tatgctccat ccataatctt catcgatgag attgacacac ttggtggaca acgtggaaat
     2101 tctggagaac acgaagcgag tcggcgtgtt aaatccgagt ttctggtaca aatggacggg
     2161 tcacagaata aattcgattc gcgacgtgta ttcgtgctgg ccgccacaaa tattccatgg
     2221 gagctcgatg aggcactccg aagacgtttc gaaaagcgaa tcttcattcc actgccagat
     2281 atagatgcaa gaaagaagct tatcgaaaaa tcaatggaag gaactccgaa atccgatgaa
     2341 attaactatg atgacttggc ggcaaggact gaaggattct caggagctga tgttgtgtct
     2401 ctatgcagaa ctgctgctat caatgtgttg agaagatatt tcagatacga caccaaatca
     2461 ttgcgaggcg gggaactgac agctgcaatg gaatcactga aagccgaact cgtgaggaac
     2521 attgactttg aagcagctct tcaagcagtt tcaccaagtg ctggtccaga tacaatgctc
     2581 aaatgcaaag aatggtgtga ctcttttggt gccatgtaag acccagcttt cttgtacaaa
     2641 gtggtgatat ttcagaattc ggatcctcta gagtcgacct gcaggcaagc ttggcactgg
     2701 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     2761 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     2821 cccaacagtt gcgcagcctg aatggcgaat ggcagcttgg ctgttttggc ggatgagata
     2881 agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt
     2941 tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac
     3001 gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat
     3061 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     3121 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
     3181 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag
     3241 aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttttgtt tatttttcta
     3301 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3361 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3421 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3481 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3541 tgagagtttt cgccccgaag aacgtttccc aatgatgagc acttttaaag ttctgctatg
     3601 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3661 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3721 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3781 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3841 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3901 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     3961 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4021 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4081 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4141 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4201 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4261 tatactttag attgatttac cccggttgat aatcagaaaa gccccaaaaa caggaagatt
     4321 gtataagcaa atatttaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt
     4381 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4441 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     4501 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     4561 cgtgaaccat cacccaaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     4621 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4681 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     4741 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtaaaaggat
     4801 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4861 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4921 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4981 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5041 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5101 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5161 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5221 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5281 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5341 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5401 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5461 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5521 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5581 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     5641 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     5701 gcatctgtgc ggtatttcac accgcatata tggtgcactc tcagtacaat ctgctctgat
     5761 gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc
     5821 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     5881 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     5941 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtgc agcgattcac
     6001 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     6061 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct
     6121 ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc
     6181 tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac
     6241 aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct
     6301 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
     6361 ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac
     6421 cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg
     6481 ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc
     6541 gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ccaggaccca acgctgcccg
     6601 aaattccgac accatcgaat ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga
     6661 agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta
     6721 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc
     6781 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt
     6841 ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc
     6901 cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc
     6961 cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca
     7021 caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga
     7081 tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga
     7141 ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga
     7201 gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt
     7261 ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc
     7321 gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat
     7381 gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg
     7441 cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg
     7501 atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga
     7561 ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc
     7621 ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc
     7681 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     7741 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc
     7801 attaggcaca attctcatgt ttgacagctt atcatcgact gcacggtgca ccaatgcttc
     7861 tggcgtcagg cagccatcgg aagctgtggt atggctgtgc aggtcgtaaa tcactgcata
     7921 attcgtgtcg ctcaaggcgc actcccgttc tggataatgt tttttgcgcc gacatcataa
     7981 cggttctggc aaatattctg aaatgagctg ttgacaatta atcatcggct cgtataatgt
     8041 gtggaattgt gagcggataa caatttcaca caggaaacag ccagtccgtt taggtgtttt
     8101 cacgagcact tcaccaacaa ggaccatagc at
//
