LOCUS       sequence-194528-        9599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for expression of NHA-tagged Drosophila melanogaster GW182 
            6W6A with six mutated W-motifs in its N-terminal domain in D. 
            melanogaster cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9599)
  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 9599)
  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-623
FEATURES             Location/Qualifiers
     source          1..9599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1857..1922
                     /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"
     CDS             1950..1976
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6176..6217
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6227..6244
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6267..6284)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6317..6451
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6341..6360)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6395..6414
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7052..7640)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7132..7151)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7811..8602,8603..8671))
                     /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     8434..8453
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8672..8776)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8844..8862
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(8926..9599,1..1836)
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
ORIGIN
        1 gatgatgcca gaccaaaagg cttgaatttc tgcgtctttt gccgaacgca gtgcatgtgc
       61 aattgttgtt ttttggaata ttcaattttc ggactgtccg ctttgatttc agtttcttgg
      121 cttattcaaa aagcaaagta aagccaaaaa agcgagatgg caataccaaa tgcggcaaaa
      181 cggtagtgga aggaaagggg tgcggggcag cggaaggaag ggtggggcgg ggcgtggcgg
      241 ggtctgtggc tgggcgcgac gtcaccgacg ttggagccac tcctttgacc atgtgtgcgt
      301 gtgtgtatta ttcgtgtctc gccactcgcc ggttgttttt ttctttttat gctgcgctct
      361 ctctagcgcc atctcgctta cgcatgctca acgcaccgca tgttgccgtt tccttttatg
      421 cgtcattttg gctcgaaata ggcaattatt taaacaaaga ttagtcaacg aaaacgctaa
      481 aataaataag tctacaatat ggttacttat tgccatgtgt gtgcagccaa cgatagcaac
      541 aaaagcaaca acacaggtgg ctttccctct ttcacttttt gtttgcaagc cgcgtgcgag
      601 caagacggca cgaccggcaa acgcaattac gctgacaaag agcagacgaa gttttggcga
      661 aaaacatcaa ggcgcctgat acgaatgcat ttgcaataac aattgcgata tttaatattg
      721 tttatgaagc tgtttgactt caaaacacac aaaaaaaaaa ataaaacaaa ttatttgaaa
      781 gagaattagg aatcggacgc ttatcgttag ggtaacaaca agaaatgctt actgagtcac
      841 agcctctgga aaactgccgc aagccagaga gagagagaaa aagagggaga gcagcttaga
      901 ccgcatgtgc ttgtgtgtga ggcgtctctc tcttcgtctc tgttgcgcaa acgcatagac
      961 tgcactgaaa aaatcgatta cctatttttt atgaatgaat atttgcacta ttactattca
     1021 aaactattaa gatagcaatc acattcaata gccaaatact ataccacctg agcgatgcaa
     1081 cgaaatgatc aatttgagca aaaatgctgc atatttagga cggcatcatt atagaaatgc
     1141 ttcttgctgt gtacttttct ctcgtctggc agctgtttcg ccgttattgt taaaaccggc
     1201 ttaagttagg tgtgttttct acgactagtg aatgccctac tagaagatgt gtgttgcaca
     1261 aaatgtccct ggaataacca atttgaagtg cagatagcag taaacgtaag ctaatatgaa
     1321 tattatttaa ctgtaatgtt ttaatatcgc tggacattac taataaaccc actataaaca
     1381 catgtacata tgtatgtttt ggcatacaat gagtagttgg ggaaaaaatg tgtaaaagca
     1441 ccgtgaccat cacagcataa agataaccag ctgaagtatc gaatatgagt aacccccaaa
     1501 ttgaatcaca tgccgcaact gataggaccc atggaagtac actcttcatg gcgatataca
     1561 agacacacac aagcacgaac acccagttgc ggaggaaatt ctccgtaaat gaaaacccaa
     1621 tcggcgaaca attcataccc atatatggta aaagttttga acgcgacttg agagcggaga
     1681 gcattgcggc tgataaggtt ttagcgctaa gcgggcttta taaaacgggc tgcgggacca
     1741 gttttcatat cactaccgtt tgagttcttg tgctgtgtgg atactcctcc cgacacaaag
     1801 ccgctccatc agccagcagt cgtctaatcc agagaccccg gatcggggta cccaacatgg
     1861 acgcacaaac acgacgacgt gagcgtcgcg ctgagaaaca agctcaatgg aaagctgcaa
     1921 acggtggatc tcctccaagc ttgatggctt acccatacga tgttccagat tacgctcctg
     1981 caggcatgcg tgaagccctt ttttcccaag atggctgggg ctgtcagcat gttaaccagg
     2041 atactaattg ggaagttccc agttcgccag aaccagccaa taaggatgca cccggtccac
     2101 caatgtggaa gccaagcatt aacaatggta ctgatctttg ggagtccaat ttgagaaacg
     2161 gaggtcagcc ggccgcacag caagttccaa agccgtcgtg gggtcataca ccatcctcta
     2221 acttaggtgg aacatggggt gaggacgacg atggcgccga tagtagtagt gtgtggactg
     2281 gaggagctgt tagcaacgcg ggatccggag ctgcagtggg agtaaaccaa gccggagtta
     2341 atgtcggtcc aggcggtgtt gtttcgtctg gcggacctca gtggggacaa ggtgtcgttg
     2401 gcgtcggact tggatcaact ggaggtaacg ggtcaagcaa tataactgga tcgtctggag
     2461 tcgcaacagg tagtagcgga aactccagca acgctggtaa cggttgggga gaccctcgtg
     2521 aaatacgccc tttgggagtt ggtggctcca tggatattcg aaatgttgaa catcgcggcg
     2581 gtaacggttc tggagcaact tcgagcgatc cacgagacat tcgcatgatc gatccgcgtg
     2641 accctattcg aggagatccc cgtggaatat ctggtcgtct taatgggacc tctgaaatgg
     2701 cgggtcatca tccacaaatg tcccataacc agttgcaagg tatcaacaaa atggttggtc
     2761 aaagtgtagc aactgccagc accagtgtcg gaacatctgg ctcgggcatc ggtcctggag
     2821 gtcccggtcc tagtacagta tcaggcaata tcccaacaca ggcggggcct gctcaaccgg
     2881 taagcgttgg tgtaagtggt cccaaagaca tgtcaaaaca gataagtgga gcggaggaac
     2941 catcaccacc gcctcagcgt cgcagtattc ctaactacga tgatggtaca tcgttggcgg
     3001 gtcagcaaac tcgtgttccc gctgcaagcg gtcactggaa agacatgact gattcgatag
     3061 gtcgtagtag tcatctcatg cgtggccaaa gccaaacggg aggtatagga atagccggcg
     3121 ttggaaatag caatgttcca gtgggagcca atccaagtaa tcctataagc agtgtagttg
     3181 gacctcaagc ccggattcca tctgtgggcg gcgtacaaca caaaccagac ggcggcgcta
     3241 tgtgggtgca ttccggcaat gtaggtggca gaaataatgt tgctgctgtt actactgcgg
     3301 gagatgacac tcatagcgtt aatgtcggcg ctcccagcag tggcagtgta tccagcaaca
     3361 attgggttga tgacaagtcc aactcaacct tggcacaaaa ctcttggagc gacccggccc
     3421 ctgttggagt tagtgcgggc aataagcaaa gcaaaccgcc aagcaatagt gcttcatcag
     3481 gttggagcac tgctgcgggc gtggtggatg gggttgatct aggatctgag tggaacacgc
     3541 acggggggat tattggaaaa tctcagcaac aacaaaaact agcgggactt aacgtgggaa
     3601 tggtgaacgt aattaacgcg gagatcatta agcaaagcaa gcaatacagg atccttgtcg
     3661 agaacggctt taaaaaggaa gatgtagagc gggcattagt gattgctaat atgaacatcg
     3721 aagaggcagc cgatatgctc cgtgccaact catccctatc aatggatggt tggcgtcgac
     3781 atgatgagtc ccttggatct tatgccgacc acaatagttc aacaagcagc ggtggatttg
     3841 ctggtcgtta cccggtcaac agtggacaac cttcaatgtc ctttcctcat aataacctta
     3901 tgaataacat gggaggtacc gctgttactg gaggtaacaa caatacaaac atgacagctt
     3961 tacaggtgca aaagtattta aatcaagggc aacatggtgt cgctgttgga ccgcaagccg
     4021 ttggtaattc ttcagcagta tctgtcggat ttggtcagaa cacgtctaac gcagcagtgg
     4081 caggagcagc ctctgcaaat atagcagcaa atacaaacaa ccaaccgtct ggtcagcaaa
     4141 ttcgcatgct aggccagcaa attcagttgg ccattcatag tggtttcata tctagtcaga
     4201 tattgactca accgctaact caaacaaccc ttaacctttt aaaccaactt cttagcaata
     4261 ttaagcatct ccaggctgca cagcaatccc ttacccgcgg gggaaatgtc aatccaatgg
     4321 cagtgaatgt ggctatatct aaatacaagc agcaaatcca gaatttacag aaccagataa
     4381 gtgcacaaca ggctgtgtat gtaaaacagc aaaatatgca accaacttca caacaacaac
     4441 agccccaaca acagcaactt ccttctgttc atctaagtaa ctcaggcaac gactatttaa
     4501 gaggtcacga tgcaataaat aatttgcaaa gcaacttttc tgagctcaat attaataagc
     4561 caagtggata tcaaggagcg tccaatcaac aatcccgatt aaatcagtgg aagcttccag
     4621 tattagataa ggagatcaac tctgacagta cggaattttc tcgtgcccca ggtgcaacga
     4681 aacaaaattt gacggccaac acaagcaaca taaactcttt gggtcttcaa aacgatagta
     4741 catggtcaac tggacgcagt attggtgacg gttggcctga tccctcatct gataacgaga
     4801 ataaagactg gtctgttgct cagccaactt cagcagcaac tgcttacact gatctggtcc
     4861 aagagtttga gccaggcaag ccatggaagg gttcacagat caaaagcata gaagatgatc
     4921 ccagcattac accaggaagc gttgctagat ctccattgtc tattaattcg acgccaaaag
     4981 atgctgacat atttgccaat accggtaaaa attcaccgac tgatttaccg ccactaagtt
     5041 tatcgtcgtc tacatggagt tttaatccaa accaaaatta tccgagtcac agttggtctg
     5101 acaatagtca acaatgtacc gccacttcgg agctttggac aagcccgcta aataaatcat
     5161 cgtctcgagg tcccccgcca ggattgactg ccaattcaaa taaatctgca aatagtaatg
     5221 cgtcaacgcc aacaactatt accggaggtg cgaatggatg gttacagcct cgaagtggcg
     5281 gtgttcaaac cacaaacact aattggacag gtggtaacac cacttggggc tccagttggt
     5341 tgcttttgaa aaatctaaca gcacagattg atggtcctac tttgcgtaca ctgtgtatgc
     5401 agcatgggcc ccttgtcagc tttcacccgt atttgaacca aggaattgcc ttatgtaaat
     5461 atactactcg tgaggaggcg aacaaggcgc aaatggcgtt aaacaactgt gtcctcgcca
     5521 acaccacaat atttgctgaa tctcccagcg agaacgaggt gcaaagcatt atgcagcact
     5581 taccacaaac tccttcctct acaagctcta gtggaactag tggtggcaac gtcggaggcg
     5641 tcggcacttc agccaataat gcaaacagtg gttctgcagc ttgtctgtcc ggaaacaata
     5701 gcggcaacgg aaacggcagc gcgagcggcg ccggcagcgg caacaatggc aacagtagct
     5761 gcaacaacag tgccgccggg gggggcagca gcagcaacaa cacgattacc actgtagcaa
     5821 attcgaatct tgttggttct agtggctctg tctcaaattc ctctggcgtt actgctaact
     5881 ctagtactgt ttctgtagtt agttgtacag cgagtgggaa ttccataaat ggggcaggta
     5941 ctgcaaacag ttctggttca aagagtagtg caaacaattt agctagcggc cagtctagcg
     6001 cttctaactt aactaatagc accaattcaa catggcgaca aactagccaa aaccaagctc
     6061 ttcaaagtca aagcaggcca tcaggcagag aagctgactt tgattatata tctctcgttt
     6121 attccattgt tgatgattaa gcggccgctc gagtctagag ggcccgcggt tcgaaggtaa
     6181 gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggtcatc atcaccatca
     6241 ccattgagtt taaacccgct gatcagcctc gactgtgcct tctaaggcct gagctcgctg
     6301 atcagcctcg atcgaggatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     6361 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     6421 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     6481 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     6541 tatggctgat tatgatcagt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
     6601 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     6661 aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct
     6721 ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc
     6781 aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg
     6841 gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga ggctggatgg
     6901 ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca
     6961 tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggccag caaaaggcca
     7021 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     7081 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     7141 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7201 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7261 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7321 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7381 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7441 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     7501 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7561 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7621 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7681 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7741 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7801 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7861 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7921 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7981 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     8041 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     8101 gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta
     8161 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     8221 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8281 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8341 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8401 gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt
     8461 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8521 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8581 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8641 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8701 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8761 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     8821 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaagaat
     8881 taattctata ttctaaaaac acaaatgata cttctaaaaa aaaatcatga atggcatcaa
     8941 ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac atcatttttc
     9001 cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg ccattaaagc
     9061 tggctgatcg tcgccaaaca ccaaatacat aatgaatatg tacacattcg agaaagaagc
     9121 gatcaaagaa gcgtcttcgg gcggagtagg agaatgcgga ggagaaggag aacgagctga
     9181 tctagtatct ctccacaatc caatgccaac tgaccaactg gccatattcg gagcaatttg
     9241 aagccaattt ccatcgcctg gcgatcgctc cattcttggc tatatgtttt tcaccgttac
     9301 ccggggccat tttcaaagac tcgtcggcaa gataagattg tgtcactcgc tgtctctctt
     9361 catttgtcga agaatgctga ggaatttcgc gatgacgtcg gcgagtattt tgaagaatga
     9421 gaataatttg tatttatacg aaaatcagtt agtggaattt tctacaaaaa catgttatct
     9481 atagataatt ttgttgcaaa atatgttgac tatgacaaag attgtatgta tataccttta
     9541 atgtattctc attttcttat gtatttataa tggcaatgat gatactgatg atattttaa
//
