LOCUS       sequence-260048-        6941 bp    DNA     circular SYN 15-APR-2025
DEFINITION  expression of MEG-3IDR (aa1-544) in C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6941)
  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 6941)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pDC21
FEATURES             Location/Qualifiers
     source          1..6941
                     /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     732..749
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    764..903
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(829..848)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     989..1011
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1005..1196)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2005..3087)
                     /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     3049..3068
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3088..3165)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3371..3390
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3474..3493
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3474..3492
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3493..3517
                     /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             3532..3554
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3573..3590
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3600..3617
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5290..5307
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5360..5378)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5374..5421
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5458..5913
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5545..5564)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5755..5776
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6005..6820)
                     /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(6005..6820)
                     /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     6728..6747
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6812..6831
                     /label=pENTR-R
                     /note="pENTR vectors, 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 ctcacatgtt ctttcctgcg ttatcccctg
      661 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
      721 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
      781 tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc
      841 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
      901 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
      961 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     1021 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
     1081 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
     1141 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat
     1201 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
     1261 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
     1321 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
     1381 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
     1441 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
     1501 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     1561 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     1621 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg
     1681 ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg
     1741 agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga
     1801 aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg
     1861 ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag
     1921 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     1981 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2041 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     2101 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     2161 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     2221 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     2281 cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     2341 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     2401 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     2461 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     2521 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     2581 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     2641 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     2701 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     2761 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     2821 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     2881 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     2941 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     3001 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     3061 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     3121 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     3181 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     3241 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     3301 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     3361 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     3421 tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg
     3481 actcactata ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa
     3541 ctttaagaag gagatatacc atgggcagca gccatcatca tcatcatcac agcagcggcc
     3601 tggtgccgcg cggcagccat atgagttcct caaaacctta cccaagcggt ctgccaaact
     3661 cgagaagaaa aagaggaggt cgccgctcaa gtagtcgtag caatcaggaa agtgcatcca
     3721 acaacatgga acaccaaatc actcttgacg aattgttcaa tccgatcgca aaacaagaca
     3781 gtgctcagtc aacaagccgt gaatatggtg caaaatcggg gatatctcat cacggatccg
     3841 tatcattcaa cgggaataca ttcatgaatg gccaacagct gaatcactcg atgactcgac
     3901 atggtcgagt attcaatcag tcgatgcatg cagctcaagg aaacggcagc aacgccttta
     3961 acagtattcc tcctacggcc cccgtcttct ctgcagactt ccgccgcaac ttgcaaacac
     4021 gcaatagctc cagctggtat gaacgaagat tcccagtatc cacagatcag gacgacgttc
     4081 agcaatcaaa cacaagacgt agtagaagtc gtcagaatgg acaacatggc ctaagcttct
     4141 cggatggttc gaacaattat ggtcacgctg ggaataagtc attcagtgtg agttccgttc
     4201 ccgttgggtt ccagaagcaa gaaaataatt ctaagaaact gcgacaaacc aatgttcacc
     4261 agcaatgtct tggaaataaa agcttcaacg ctcaagctgg agttcatgga cacgctttta
     4321 aaaaaggcca taaggacaat aaaaatgcat ctggaaagga ggttatcaat tctagtcttg
     4381 tccaaaagca cgatgctatt aagtcgcgga atttgaacca gagcttctct ggatttccta
     4441 cacatgaaac ctcatcgatg aaaaatcaac aacagaaatc aagaaacgac agaaaaaaat
     4501 cacgtggtag cagcaacttc caggatcgta cttacttcaa cacaaatgac gatgaattaa
     4561 ctgacgatgt gttcatagac gattccatgg atgctgcccg tggtcggaga tctcgatcag
     4621 tcactaagaa acttcaacaa tcgacttatt caaagcagaa tgcaggcagt aaacaattga
     4681 ctgaaaaatg caaatcttct gaagaagcgg ctaaacgtaa cctggtatcc aatgtattca
     4741 gcaaggacgg cactgaactc tccattgaac agttattgga aattgtcagc atgaagatcg
     4801 gacaacagat tcacttgcct tccagctctc atggagaatg cagcaatttg aatcggacat
     4861 taccagcaag tgatctaaat tgctcaattg gtgaagactt tgactccagt tttgtggatg
     4921 ctaacaacca aaccctacca gtgagtcttc ccaagaaaac ttcactgtcg atcaagcgcc
     4981 gtggaagttc tcgtagtgcc tcccgtctgg ccagtcttga tgtgacactg gaaactgttg
     5041 aagaagacga ggaacccacg ccaagtccac agccgagttc tcctccaaag atctcaagaa
     5101 gaaagtggac aggaactttt gatgccaacg ttgaggaaat gagacgttta cttcatgggg
     5161 accctgagat gccgaaatct gcaaatcgtg cttcctcaag caaggatcag atcaatcgga
     5221 acaatgtgga tgtaaagaga acaccttcct cgtgagagct ccgtcgacaa gcttgcggcc
     5281 gcactcgagc accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa
     5341 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     5401 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg
     5461 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5521 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     5581 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5641 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     5701 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5761 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     5821 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5881 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     5941 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta
     6001 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     6061 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     6121 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     6181 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     6241 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     6301 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     6361 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     6421 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     6481 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     6541 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     6601 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     6661 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     6721 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     6781 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     6841 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc
     6901 cactgagcgt cagaccccgt agaaaagatc aaaggatctt c
//
