LOCUS       sequence-194379-        6275 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for expression of NHA-tagged Drosophila melanogaster GW182 
            CED 7W7A with seven mutated W-motifs in D. melanogaster cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6275)
  AUTHORS   Chekulaeva M, Mathys H, Zipprich JT, Attig J, Colic M, Parker R, 
            Filipowicz W
  TITLE     miRNA repression involves GW182-mediated recruitment of CCR4-NOT 
            through conserved W-containing motifs.
  JOURNAL   Nat Struct Mol Biol. 2011 Oct 7;18(11):1218-26. doi: 
            10.1038/nsmb.2166.
  PUBMED    21984184
REFERENCE   2  (bases 1 to 6275)
  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"; doi: "10.1038/nsmb"; 
            journalName: "Nat Struct Mol Biol"; date: "2011-10-7- 7"; volume: 
            "18"; issue: "11"; pages: "1218-26"
COMMENT     Sequence Label: pMCh-646
FEATURES             Location/Qualifiers
     source          1..6275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(966..984)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1052..1156
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1157..1225,1226..2017)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1375..1394)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2188..2776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2677..2696
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3377..3511
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3414..3433)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3468..3487
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3544..3561
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3584..3601)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3611..3652)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(4528..4554)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(4582..4647)
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     promoter        complement(join(4668..6275,1..902))
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
ORIGIN
        1 gccccaccct tccttccgct gccccgcacc cctttccttc cactaccgtt ttgccgcatt
       61 tggtattgcc atctcgcttt tttggcttta ctttgctttt tgaataagcc aagaaactga
      121 aatcaaagcg gacagtccga aaattgaata ttccaaaaaa caacaattgc acatgcactg
      181 cgttcggcaa aagacgcaga aattcaagcc ttttggtctg gcatcatctt aaaatatcat
      241 cagtatcatc attgccatta taaatacata agaaaatgag aatacattaa aggtatatac
      301 atacaatctt tgtcatagtc aacatatttt gcaacaaaat tatctataga taacatgttt
      361 ttgtagaaaa ttccactaac tgattttcgt ataaatacaa attattctca ttcttcaaaa
      421 tactcgccga cgtcatcgcg aaattcctca gcattcttcg acaaatgaag agagacagcg
      481 agtgacacaa tcttatcttg ccgacgagtc tttgaaaatg gccccgggta acggtgaaaa
      541 acatatagcc aagaatggag cgatcgccag gcgatggaaa ttggcttcaa attgctccga
      601 atatggccag ttggtcagtt ggcattggat tgtggagaga tactagatca gctcgttctc
      661 cttctcctcc gcattctcct actccgcccg aagacgcttc tttgatcgct tctttctcga
      721 atgtgtacat attcattatg tatttggtgt ttggcgacga tcagccagct ttaatggcca
      781 taaaacgtgt cttggtttgt gggccacata acaggcagcg agatctggaa aaatgatgtg
      841 acagtggaaa tgagaagtag gtgcatctgc aaagatttga ttcagagttg atgccattca
      901 tgattttttt ttagaagtat catttgtgtt tttagaatat agaattaatt cttgaagacg
      961 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     1021 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1081 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1141 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1201 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1261 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     1321 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     1381 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     1441 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     1501 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     1561 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     1621 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     1681 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     1741 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     1801 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     1861 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     1921 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     1981 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2041 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2101 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2161 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2221 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2281 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     2341 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2401 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2461 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2521 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2581 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2641 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2701 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2761 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2821 gccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc
     2881 gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca tccagcctcg
     2941 cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg cgataatggc
     3001 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     3061 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     3121 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     3181 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     3241 gttgaaggct ctcaagggca tcggtcgact gatcataatc agccatacca catttgtaga
     3301 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     3361 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3421 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3481 actcatcaat gtatcttatc atgtctggat cctcgatcga ggctgatcag cgagctcagg
     3541 ccttagaagg cacagtcgag gctgatcagc gggtttaaac tcaatggtga tggtgatgat
     3601 gaccggtacg cgtagaatcg agaccgagga gagggttagg gataggctta ccttcgaacc
     3661 gcgggccctc tagactcgag cggccgctta gttgccacca ctagttccac tagagcttgt
     3721 agaggaagga gtttgtggta agtgctgcat aatgctttgc acctcgttct cgctgggaga
     3781 ttcagcaaat attgtggtgt tggcgaggac acagttgttt aacgccattt gcgccttgtt
     3841 cgcctcctca cgagtagtat atttacataa ggcaattcct tggttcaaat acgggtgaaa
     3901 gctgacaagg ggcccatgct gcatacacag tgtacgcaaa gtaggaccat caatctgtgc
     3961 tgttagattt ttcaaaagca accaactgga gcccgcagtg gtgttaccac ctgtcgcatt
     4021 agtgtttgtg gtttgaacac cgccacttcg aggctgtaat gctccattcg cacctccggt
     4081 aatagttgtt ggcgttgacg cattactatt tgcagattta tttgaattgg cagtcaatcc
     4141 tggcggggga cctcgagacg atgatttatt tagcgggctt gttgcaagct ccgaagtggc
     4201 ggtacattgt tgactattgt cagatgcact gtgactcgga taattttggt ttggattaaa
     4261 acttgctgta gacgacgata aacttagtgg cggtaaatca gtcggtgaat ttttaccggt
     4321 attggcaaat atgtcagcat cttttggcgt cgaattaata gacaatggag atctagcaac
     4381 gcttcctggt gtaatgctgg gatcatcttc tatgcttttg atctgtgaac ccttccatgg
     4441 cttgcctggc tcaaactctt ggaccagatc agtgtaagca gttgctgctg aagttggctg
     4501 agcaacagat gcgtctttgc ctgcaggagc gtaatctgga acatcgtatg ggtaagccat
     4561 caagcttgga ggagatccac cgtttgcagc tttccattga gcttgtttct cagcgcgacg
     4621 ctcacgtcgt cgtgtttgtg cgtccatgtt gggtaccccg atccggggtc tctggattag
     4681 acgactgctg gctgatggag cggctttgtg tcgggaggag tatccacaca gcacaagaac
     4741 tcaaacggta gtgatatgaa aactggtccc gcagcccgtt ttataaagcc cgcttagcgc
     4801 taaaacctta tcagccgcaa tgctctccgc tctcaagtcg cgttcaaaac ttttaccata
     4861 tatgggtatg aattgttcgc cgattgggtt ttcatttacg gagaatttcc tccgcaactg
     4921 ggtgttcgtg cttgtgtgtg tcttgtatat cgccatgaag agtgtacttc catgggtcct
     4981 atcagttgcg gcatgtgatt caatttgggg gttactcata ttcgatactt cagctggtta
     5041 tctttatgct gtgatggtca cggtgctttt acacattttt tccccaacta ctcattgtat
     5101 gccaaaacat acatatgtac atgtgtttat agtgggttta ttagtaatgt ccagcgatat
     5161 taaaacatta cagttaaata atattcatat tagcttacgt ttactgctat ctgcacttca
     5221 aattggttat tccagggaca ttttgtgcaa cacacatctt ctagtagggc attcactagt
     5281 cgtagaaaac acacctaact taagccggtt ttaacaataa cggcgaaaca gctgccagac
     5341 gagagaaaag tacacagcaa gaagcatttc tataatgatg ccgtcctaaa tatgcagcat
     5401 ttttgctcaa attgatcatt tcgttgcatc gctcaggtgg tatagtattt ggctattgaa
     5461 tgtgattgct atcttaatag ttttgaatag taatagtgca aatattcatt cataaaaaat
     5521 aggtaatcga ttttttcagt gcagtctatg cgtttgcgca acagagacga agagagagac
     5581 gcctcacaca caagcacatg cggtctaagc tgctctccct ctttttctct ctctctctgg
     5641 cttgcggcag ttttccagag gctgtgactc agtaagcatt tcttgttgtt accctaacga
     5701 taagcgtccg attcctaatt ctctttcaaa taatttgttt tatttttttt tttgtgtgtt
     5761 ttgaagtcaa acagcttcat aaacaatatt aaatatcgca attgttattg caaatgcatt
     5821 cgtatcaggc gccttgatgt ttttcgccaa aacttcgtct gctctttgtc agcgtaattg
     5881 cgtttgccgg tcgtgccgtc ttgctcgcac gcggcttgca aacaaaaagt gaaagaggga
     5941 aagccacctg tgttgttgct tttgttgcta tcgttggctg cacacacatg gcaataagta
     6001 accatattgt agacttattt attttagcgt tttcgttgac taatctttgt ttaaataatt
     6061 gcctatttcg agccaaaatg acgcataaaa ggaaacggca acatgcggtg cgttgagcat
     6121 gcgtaagcga gatggcgcta gagagagcgc agcataaaaa gaaaaaaaca accggcgagt
     6181 ggcgagacac gaataataca cacacgcaca catggtcaaa ggagtggctc caacgtcggt
     6241 gacgtcgcgc ccagccacag accccgccac gcccc
//
