LOCUS       sequence-194406-        9599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for expression of NHA-tagged Drosophila melanogaster GW182 
            in D. melanogaster cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9599)
  AUTHORS   Chekulaeva M, Filipowicz W, Parker R
  TITLE     Multiple independent domains of dGW182 function in miRNA-mediated 
            repression in Drosophila.
  JOURNAL   RNA. 2009 May;15(5):794-803. doi: 10.1261/rna.1364909. Epub 2009 Mar
            20.
  PUBMED    19304924
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: 
            "10.1261/rna.1364909"; journalName: "RNA"; date: "2009-05"; volume: 
            "15"; issue: "5"; pages: "794-803"
COMMENT     Sequence Label: pMCh-61
FEATURES             Location/Qualifiers
     source          1..9599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1818..1883
                     /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             1911..1937
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6137..6178
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6188..6205
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6228..6245)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6278..6412
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6302..6321)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6356..6375
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7013..7601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7093..7112)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7772..8563,8564..8632))
                     /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     8395..8414
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8633..8737)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8805..8823
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(8887..9599,1..1797)
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
ORIGIN
        1 tgccgaacgc agtgcatgtg caattgttgt tttttggaat attcaatttt cggactgtcc
       61 gctttgattt cagtttcttg gcttattcaa aaagcaaagt aaagccaaaa aagcgagatg
      121 gcaataccaa atgcggcaaa acggtagtgg aaggaaaggg gtgcggggca gcggaaggaa
      181 gggtggggcg gggcgtggcg gggtctgtgg ctgggcgcga cgtcaccgac gttggagcca
      241 ctcctttgac catgtgtgcg tgtgtgtatt attcgtgtct cgccactcgc cggttgtttt
      301 tttcttttta tgctgcgctc tctctagcgc catctcgctt acgcatgctc aacgcaccgc
      361 atgttgccgt ttccttttat gcgtcatttt ggctcgaaat aggcaattat ttaaacaaag
      421 attagtcaac gaaaacgcta aaataaataa gtctacaata tggttactta ttgccatgtg
      481 tgtgcagcca acgatagcaa caaaagcaac aacacaggtg gctttccctc tttcactttt
      541 tgtttgcaag ccgcgtgcga gcaagacggc acgaccggca aacgcaatta cgctgacaaa
      601 gagcagacga agttttggcg aaaaacatca aggcgcctga tacgaatgca tttgcaataa
      661 caattgcgat atttaatatt gtttatgaag ctgtttgact tcaaaacaca caaaaaaaaa
      721 aataaaacaa attatttgaa agagaattag gaatcggacg cttatcgtta gggtaacaac
      781 aagaaatgct tactgagtca cagcctctgg aaaactgccg caagccagag agagagagaa
      841 aaagagggag agcagcttag accgcatgtg cttgtgtgtg aggcgtctct ctcttcgtct
      901 ctgttgcgca aacgcataga ctgcactgaa aaaatcgatt acctattttt tatgaatgaa
      961 tatttgcact attactattc aaaactatta agatagcaat cacattcaat agccaaatac
     1021 tataccacct gagcgatgca acgaaatgat caatttgagc aaaaatgctg catatttagg
     1081 acggcatcat tatagaaatg cttcttgctg tgtacttttc tctcgtctgg cagctgtttc
     1141 gccgttattg ttaaaaccgg cttaagttag gtgtgttttc tacgactagt gaatgcccta
     1201 ctagaagatg tgtgttgcac aaaatgtccc tggaataacc aatttgaagt gcagatagca
     1261 gtaaacgtaa gctaatatga atattattta actgtaatgt tttaatatcg ctggacatta
     1321 ctaataaacc cactataaac acatgtacat atgtatgttt tggcatacaa tgagtagttg
     1381 gggaaaaaat gtgtaaaagc accgtgacca tcacagcata aagataacca gctgaagtat
     1441 cgaatatgag taacccccaa attgaatcac atgccgcaac tgataggacc catggaagta
     1501 cactcttcat ggcgatatac aagacacaca caagcacgaa cacccagttg cggaggaaat
     1561 tctccgtaaa tgaaaaccca atcggcgaac aattcatacc catatatggt aaaagttttg
     1621 aacgcgactt gagagcggag agcattgcgg ctgataaggt tttagcgcta agcgggcttt
     1681 ataaaacggg ctgcgggacc agttttcata tcactaccgt ttgagttctt gtgctgtgtg
     1741 gatactcctc ccgacacaaa gccgctccat cagccagcag tcgtctaatc cagagacccc
     1801 ggatcggggt acccaacatg gacgcacaaa cacgacgacg tgagcgtcgc gctgagaaac
     1861 aagctcaatg gaaagctgca aacggtggat ctcctccaag cttgatggct tacccatacg
     1921 atgttccaga ttacgctcct gcaggcatgc gtgaagccct tttttcccaa gatggctggg
     1981 gctgtcagca tgttaaccag gatactaatt gggaagttcc cagttcgcca gaaccagcca
     2041 ataaggatgc acccggtcca ccaatgtgga agccaagcat taacaatggt actgatcttt
     2101 gggagtccaa tttgagaaac ggaggtcagc cggccgcaca gcaagttcca aagccgtcgt
     2161 ggggtcatac accatcctct aacttaggtg gaacatgggg tgaggacgac gatggcgccg
     2221 atagtagtag tgtgtggact ggaggagctg ttagcaacgc gggatccgga gctgcagtgg
     2281 gagtaaacca agccggagtt aatgtcggtc caggcggtgt tgtttcgtct ggcggacctc
     2341 agtggggaca aggtgtcgtt ggcgtcggac ttggatcaac tggaggtaac gggtcaagca
     2401 atataactgg atcgtctgga gtcgcaacag gtagtagcgg aaactccagc aacgctggta
     2461 acggttgggg agaccctcgt gaaatacgcc ctttgggagt tggtggctcc atggatattc
     2521 gaaatgttga acatcgcggc ggtaacggtt ctggagcaac ttcgagcgat ccacgagaca
     2581 ttcgcatgat cgatccgcgt gaccctattc gaggagatcc ccgtggaata tctggtcgtc
     2641 ttaatgggac ctctgaaatg tggggtcatc atccacaaat gtcccataac cagttgcaag
     2701 gtatcaacaa aatggttggt caaagtgtag caactgccag caccagtgtc ggaacatctg
     2761 gctcgggcat cggtcctgga ggtcccggtc ctagtacagt atcaggcaat atcccaacac
     2821 agtgggggcc tgctcaaccg gtaagcgttg gtgtaagtgg tcccaaagac atgtcaaaac
     2881 agataagtgg atgggaggaa ccatcaccac cgcctcagcg tcgcagtatt cctaactacg
     2941 atgatggtac atcgttgtgg ggtcagcaaa ctcgtgttcc cgctgcaagc ggtcactgga
     3001 aagacatgac tgattcgata ggtcgtagta gtcatctcat gcgtggccaa agccaaacgg
     3061 gaggtatagg aatagccggc gttggaaata gcaatgttcc agtgggagcc aatccaagta
     3121 atcctataag cagtgtagtt ggacctcaag cccggattcc atctgtgggc ggcgtacaac
     3181 acaaaccaga cggcggcgct atgtgggtgc attccggcaa tgtaggtggc agaaataatg
     3241 ttgctgctgt tactacttgg ggagatgaca ctcatagcgt taatgtcggc gctcccagca
     3301 gtggcagtgt atccagcaac aattgggttg atgacaagtc caactcaacc ttggcacaaa
     3361 actcttggag cgacccggcc cctgttggag ttagttgggg caataagcaa agcaaaccgc
     3421 caagcaatag tgcttcatca ggttggagca ctgctgcggg cgtggtggat ggggttgatc
     3481 taggatctga gtggaacacg cacgggggga ttattggaaa atctcagcaa caacaaaaac
     3541 tagcgggact taacgtggga atggtgaacg taattaacgc ggagatcatt aagcaaagca
     3601 agcaatacag gatccttgtc gagaacggct ttaaaaagga agatgtagag cgggcattag
     3661 tgattgctaa tatgaacatc gaagaggcag ccgatatgct ccgtgccaac tcatccctat
     3721 caatggatgg ttggcgtcga catgatgagt cccttggatc ttatgccgac cacaatagtt
     3781 caacaagcag cggtggattt gctggtcgtt acccggtcaa cagtggacaa ccttcaatgt
     3841 cctttcctca taataacctt atgaataaca tgggaggtac cgctgttact ggaggtaaca
     3901 acaatacaaa catgacagct ttacaggtgc aaaagtattt aaatcaaggg caacatggtg
     3961 tcgctgttgg accgcaagcc gttggtaatt cttcagcagt atctgtcgga tttggtcaga
     4021 acacgtctaa cgcagcagtg gcaggagcag cctctgcaaa tatagcagca aatacaaaca
     4081 accaaccgtc tggtcagcaa attcgcatgc taggccagca aattcagttg gccattcata
     4141 gtggtttcat atctagtcag atattgactc aaccgctaac tcaaacaacc cttaaccttt
     4201 taaaccaact tcttagcaat attaagcatc tccaggctgc acagcaatcc cttacccgcg
     4261 ggggaaatgt caatccaatg gcagtgaatg tggctatatc taaatacaag cagcaaatcc
     4321 agaatttaca gaaccagata agtgcacaac aggctgtgta tgtaaaacag caaaatatgc
     4381 aaccaacttc acaacaacaa cagccccaac aacagcaact tccttctgtt catctaagta
     4441 actcaggcaa cgactattta agaggtcacg atgcaataaa taatttgcaa agcaactttt
     4501 ctgagctcaa tattaataag ccaagtggat atcaaggagc gtccaatcaa caatcccgat
     4561 taaatcagtg gaagcttcca gtattagata aggagatcaa ctctgacagt acggaatttt
     4621 ctcgtgcccc aggtgcaacg aaacaaaatt tgacggccaa cacaagcaac ataaactctt
     4681 tgggtcttca aaacgatagt acatggtcaa ctggacgcag tattggtgac ggttggcctg
     4741 atccctcatc tgataacgag aataaagact ggtctgttgc tcagccaact tcagcagcaa
     4801 ctgcttacac tgatctggtc caagagtttg agccaggcaa gccatggaag ggttcacaga
     4861 tcaaaagcat agaagatgat cccagcatta caccaggaag cgttgctaga tctccattgt
     4921 ctattaattc gacgccaaaa gatgctgaca tatttgccaa taccggtaaa aattcaccga
     4981 ctgatttacc gccactaagt ttatcgtcgt ctacatggag ttttaatcca aaccaaaatt
     5041 atccgagtca cagttggtct gacaatagtc aacaatgtac cgccacttcg gagctttgga
     5101 caagcccgct aaataaatca tcgtctcgag gtcccccgcc aggattgact gccaattcaa
     5161 ataaatctgc aaatagtaat gcgtcaacgc caacaactat taccggaggt gcgaatggat
     5221 ggttacagcc tcgaagtggc ggtgttcaaa ccacaaacac taattggaca ggtggtaaca
     5281 ccacttgggg ctccagttgg ttgcttttga aaaatctaac agcacagatt gatggtccta
     5341 ctttgcgtac actgtgtatg cagcatgggc cccttgtcag ctttcacccg tatttgaacc
     5401 aaggaattgc cttatgtaaa tatactactc gtgaggaggc gaacaaggcg caaatggcgt
     5461 taaacaactg tgtcctcgcc aacaccacaa tatttgctga atctcccagc gagaacgagg
     5521 tgcaaagcat tatgcagcac ttaccacaaa ctccttcctc tacaagctct agtggaacta
     5581 gtggtggcaa cgtcggaggc gtcggcactt cagccaataa tgcaaacagt ggttctgcag
     5641 cttgtctgtc cggaaacaat agcggcaacg gaaacggcag cgcgagcggc gccggcagcg
     5701 gcaacaatgg caacagtagc tgcaacaaca gtgccgccgg ggggggcagc agcagcaaca
     5761 acacgattac cactgtagca aattcgaatc ttgttggttc tagtggctct gtctcaaatt
     5821 cctctggcgt tactgctaac tctagtactg tttctgtagt tagttgtaca gcgagtggga
     5881 attccataaa tggggcaggt actgcaaaca gttctggttc aaagagtagt gcaaacaatt
     5941 tagctagcgg ccagtctagc gcttctaact taactaatag caccaattca acatggcgac
     6001 aaactagcca aaaccaagct cttcaaagtc aaagcaggcc atcaggcaga gaagctgact
     6061 ttgattatat atctctcgtt tattccattg ttgatgatta agcggccgct cgagtctaga
     6121 gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg
     6181 taccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
     6241 ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga taagatacat
     6301 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     6361 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     6421 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     6481 gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgaccgatg cccttgagag
     6541 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
     6601 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt
     6661 tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg
     6721 gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg
     6781 agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt
     6841 tcgcgacgcg aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg
     6901 ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc
     6961 ttcaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7021 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7081 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7141 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     7201 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7261 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7321 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7381 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7441 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7501 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7561 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7621 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7681 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7741 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7801 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7861 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7921 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7981 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     8041 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg
     8101 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     8161 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     8221 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     8281 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     8341 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata
     8401 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     8461 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     8521 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     8581 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8641 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8701 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8761 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     8821 ggccctttcg tcttcaagaa ttaattctat attctaaaaa cacaaatgat acttctaaaa
     8881 aaaaatcatg aatggcatca actctgaatc aaatctttgc agatgcacct acttctcatt
     8941 tccactgtca catcattttt ccagatctcg ctgcctgtta tgtggcccac aaaccaagac
     9001 acgttttatg gccattaaag ctggctgatc gtcgccaaac accaaataca taatgaatat
     9061 gtacacattc gagaaagaag cgatcaaaga agcgtcttcg ggcggagtag gagaatgcgg
     9121 aggagaagga gaacgagctg atctagtatc tctccacaat ccaatgccaa ctgaccaact
     9181 ggccatattc ggagcaattt gaagccaatt tccatcgcct ggcgatcgct ccattcttgg
     9241 ctatatgttt ttcaccgtta cccggggcca ttttcaaaga ctcgtcggca agataagatt
     9301 gtgtcactcg ctgtctctct tcatttgtcg aagaatgctg aggaatttcg cgatgacgtc
     9361 ggcgagtatt ttgaagaatg agaataattt gtatttatac gaaaatcagt tagtggaatt
     9421 ttctacaaaa acatgttatc tatagataat tttgttgcaa aatatgttga ctatgacaaa
     9481 gattgtatgt atataccttt aatgtattct cattttctta tgtatttata atggcaatga
     9541 tgatactgat gatattttaa gatgatgcca gaccaaaagg cttgaatttc tgcgtcttt
//
