LOCUS       sequence-288111-        7906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of MEG-3 (aa1-862) in C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7906)
  AUTHORS   Smith J, Calidas D, Schmidt H, Lu T, Rasoloson D, Seydoux G
  TITLE     Spatial patterning of P granules by RNA-induced phase separation of 
            the intrinsically-disordered protein MEG-3.
  JOURNAL   Elife. 2016 Dec 3;5. pii: 21337. doi: 10.7554/eLife.21337.
  PUBMED    27914198
REFERENCE   2  (bases 1 to 7906)
  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: "2016-12-3"; volume: "5. pii: 21337. doi"; pages: " 
            10.7554/eLife.21337"
COMMENT     Sequence Label: pDC20
FEATURES             Location/Qualifiers
     source          1..7906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2771..2788
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2841..2859)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2855..2902
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2939..3394
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3026..3045)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3236..3257
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3486..4301)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3486..4301)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4209..4228
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4293..4312
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4423..5011
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4912..4931
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5165..5182
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5197..5336
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5262..5281)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5422..5444
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5438..5629)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6438..7520)
                     /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     7482..7501
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7521..7598)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7804..7823
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagccatatg agttcctcaa aaccttaccc aagcggtctg
      181 ccaaactcga gaagaaaaag aggaggtcgc cgctcaagta gtcgtagcaa tcaggaaagt
      241 gcatccaaca acatggaaca ccaaatcact cttgacgaat tgttcaatcc gatcgcaaaa
      301 caagacagtg ctcagtcaac aagccgtgaa tatggtgcaa aatcggggat atctcatcac
      361 ggatccgtat cattcaacgg gaatacattc atgaatggcc aacagctgaa tcactcgatg
      421 actcgacatg gtcgagtatt caatcagtcg atgcatgcag ctcaaggaaa cggcagcaac
      481 gcctttaaca gtattcctcc tacggccccc gtcttctctg cagacttccg ccgcaacttg
      541 caaacacgca atagctccag ctggtatgaa cgaagattcc cagtatccac agatcaggac
      601 gacgttcagc aatcaaacac aagacgtagt agaagtcgtc agaatggaca acatggccta
      661 agcttctcgg atggttcgaa caattatggt cacgctggga ataagtcatt cagtgtgagt
      721 tccgttcccg ttgggttcca gaagcaagaa aataattcta agaaactgcg acaaaccaat
      781 gttcaccagc aatgtcttgg aaataaaagc ttcaacgctc aagctggagt tcatggacac
      841 gcttttaaaa aaggccataa ggacaataaa aatgcatctg gaaaggaggt tatcaattct
      901 agtcttgtcc aaaagcacga tgctattaag tcgcggaatt tgaaccagag cttctctgga
      961 tttcctacac atgaaacctc atcgatgaaa aatcaacaac agaaatcaag aaacgacaga
     1021 aaaaaatcac gtggtagcag caacttccag gatcgtactt acttcaacac aaatgacgat
     1081 gaattaactg acgatgtgtt catagacgat tccatggatg ctgcccgtgg tcggagatct
     1141 cgatcagtca ctaagaaact tcaacaatcg acttattcaa agcagaatgc aggcagtaaa
     1201 caattgactg aaaaatgcaa atcttctgaa gaagcggcta aacgtaacct ggtatccaat
     1261 gtattcagca aggacggcac tgaactctcc attgaacagt tattggaaat tgtcagcatg
     1321 aagatcggac aacagattca cttgccttcc agctctcatg gagaatgcag caatttgaat
     1381 cggacattac cagcaagtga tctaaattgc tcaattggtg aagactttga ctccagtttt
     1441 gtggatgcta acaaccaaac cctaccagtg agtcttccca agaaaacttc actgtcgatc
     1501 aagcgccgtg gaagttctcg tagtgcctcc cgtctggcca gtcttgatgt gacactggaa
     1561 actgttgaag aagacgagga acccacgcca agtccacagc cgagttctcc tccaaagatc
     1621 tcaagaagaa agtggacagg aacttttgat gccaacgttg aggaaatgag acgtttactt
     1681 catggggacc ctgagatgcc gaaatctgca aatcgtgctt cctcaagcaa ggatcagatc
     1741 aatcggaaca atgtggatgt aaagagaaca ccttcctcgt caatcatccc aactccaaag
     1801 gctttgatcg gagaaagatg tctgacatca tccagcaagt cttcaaagtt gaacaagtca
     1861 cttggcgtgg ttgattctaa agctacaaaa tcaccaatgt attccgtcac agtttctgga
     1921 aaagaaacag cttcaggaaa gcgaattgct caaaagttga ctccaaaggt tgtggcgctg
     1981 gaaagctctt acattaccgg aattccagtt tccacggatt gtaatggttg cccaactccg
     2041 aagagaagtg gaatcaattg tgaaattcgt gcagctgaag tatacaatca agctgggaaa
     2101 tggcccttcg aaatcaccag tgatccggct ccacttcctt gtgaatcagc cgatcggatc
     2161 gaatatcctt ctcaagattg cactcaagat ccagcttcaa cctcgccacc acctcgcatt
     2221 tctgaaagct tgacagcatt cctggaagct cagcaagact ttaacgacta cattgatact
     2281 aactacaaag aaaagacaca actcctgaaa gtgaatttga atattcacgg gatgtcacct
     2341 gaaagatggc tgtacttgaa ttatttttgc accgagacaa ttccaagatt ggatggtcct
     2401 tatgccgatg atccaagagt tccacctgtg cgaaatatgt tcagaaaatg gttcctccgt
     2461 tttgctgaag cctgcttagg aaatcctcac cagcttgcag ttatgcaaga gattgctgcc
     2521 acgtttgttc aggctcggct ggacgatacg agcagctcca cagactccac caatatgctc
     2581 tacatgttgt ggaaggagtg tatcggacag aagaatatca tcgcaattgc tgatgcgtgt
     2641 ttacttgcac atcttcggaa atcggatcct atcaaatact tgaatgtaaa acgggattgg
     2701 ctggaatcta tttttgaccc acccagagat caatgagagc tccgtcgaca agcttgcggc
     2761 cgcactcgag caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga
     2821 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     2881 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
     2941 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3001 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3061 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3121 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3181 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3241 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3301 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3361 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     3421 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt
     3481 aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     3541 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     3601 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     3661 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     3721 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
     3781 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     3841 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     3901 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     3961 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     4021 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     4081 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     4141 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     4201 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     4261 cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     4321 tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg agttttcgtt
     4381 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4441 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4501 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4561 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4621 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4681 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4741 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4801 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4861 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4921 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4981 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5041 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5101 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     5161 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     5221 gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc
     5281 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     5341 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     5401 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     5461 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     5521 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     5581 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     5641 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     5701 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     5761 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     5821 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     5881 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     5941 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     6001 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     6061 tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg
     6121 cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt
     6181 gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt
     6241 cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga
     6301 agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg
     6361 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     6421 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6481 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     6541 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     6601 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     6661 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata
     6721 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     6781 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     6841 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     6901 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     6961 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     7021 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     7081 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     7141 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     7201 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     7261 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     7321 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     7381 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     7441 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     7501 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     7561 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     7621 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     7681 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     7741 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     7801 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     7861 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaat
//
