LOCUS       sequence-180802-        9525 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for expression of HA-tagged Drosophila melanogaster GW182 in
            D. melanogaster cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9525)
  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 9525)
  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-56
FEATURES             Location/Qualifiers
     source          1..9525
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2568
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             2588..2614
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6810..6851
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6861..6878
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6901..6918)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6951..7085
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6975..6994)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7029..7048
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7707..8295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7787..7806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8466..9257,9258..9326))
                     /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     9089..9108
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9327..9431)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9499..9517
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcaaggaa ttaattctat attctaaaaa cacaaatgat acttctaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat atcaatatgt acattcgaga
      241 aagaagcgat caaagaagcg tcttcgggcg agtaggagaa tgcggaggag aaggagaacg
      301 agctgatcta gtatctctcc acaatccaat gccaactgac caactggcca tattcggagc
      361 aatttgaagc caatttccat cgcctggcga tcgctccatt cttggctata tgtttttcac
      421 cgttcccggg gccattttca aagactcgtc ggtaagataa gattgtgtca ctcgctgtct
      481 ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag tattttgaag
      541 aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac aaaaacatgt
      601 tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt atgtatatac
      661 ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac tgatgatatt
      721 ttaagatgat gccagaccac aggctgattt ctgcgtcttt tgccgaacgc agtgcatgtg
      781 cggttgttgt tttttggaat agtttcaatt ttcggactgt ccgctttgat ttcagtttct
      841 tggcttattc aaaaagcaaa gtaaagccaa aaaagcgaga tggcaatacc aaatgcggca
      901 aaacggtagt ggaaggaaag gggtgcgggg cagcggaagg aagggtgggg cggggcgtgg
      961 cggggtctgt ggctgggcgc gacgtcaccg acgttggagc cactcctttg accatgtgtg
     1021 cgtgtgtgta ttattcgtgt ctcgccactc gccggttgtt tttttctttt tatctcgctc
     1081 tctctagcgc catctcgtac gcatgctcaa cgcaccgcat gttgccgtgt cctttatgcg
     1141 tcattttggc tcgaaatagg caattattta aacaaagatt agtcaacgaa aacgctaaaa
     1201 taaataagtc tacaatatgg ttacttattg ccatgtgtgt gcagccaacg atagcaacaa
     1261 aagcaacaac acagtggctt tccctctttc actttttgtt tgcaagcgcg tgcgagcaag
     1321 acggcacgac cggcaaacgc aattacgctg acaaagagca gacgaagttt tggccgaaaa
     1381 acatcaaggc gcctgatacg aatgcatttg caataacaat tgcgatattt aatattgttt
     1441 atgaagctgt ttgacttcaa aacacacaaa aaaaaaaata aaacaaatta tttgaaagag
     1501 aattaggaat cggacagctt atcgttacgg gctaacagca caccgagacg aaatagctta
     1561 cctgacgtca cagcctctgg aagaactgcc gccaagcaga gagagagaga aaaagaggga
     1621 gagcagctta gaccgcatgt gcttgtgtgt gaggcgtctc tctcttcgtc tcctgtttgc
     1681 gcaaacgcat agactgcact gagaaaatcg attacctatt ttttatgaat gaatatttgc
     1741 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca
     1801 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat
     1861 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta
     1921 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgatgccc ctactagaag
     1981 atgtgtgttg cacaaatgtc cctgaataac caatttgaag tgcagatagc agtaaacgta
     2041 agctaatatg aatattattt aactgtaatg ttttaatatc gctggacatt actaataaac
     2101 ccactataaa cacatgtaca tatgtatgtt ttggcataca atgagtagtt ggggaaaaaa
     2161 tgtgtaaaag caccgtgacc atcacagcat aaagataacc agctgaagta tcgaatatga
     2221 gtaaccccca aattgaatca catgccgcaa ctgataggac ccatggaagt acactcttca
     2281 tggcgatata caagacacac acaagcacga acacccagtt gcggaggaaa ttctccgtaa
     2341 atgaaaaccc aatcggcgaa caattcatac ccatatatgg taaaagtttt gaacgcgact
     2401 tgagagcgga gagcattgcg gctgataagg ttttagcgct aagcgggctt tataaaacgg
     2461 gctgcgggac cagttttcat atcactaccg tttgagttct tgtgctgtgt ggatactcct
     2521 cccgacacaa agccgctcca tcagccagca gtcgtctaat ccagagaccc cggatcgggt
     2581 gatggcttac ccatacgatg ttccagatta cgctcctgca ggcatgcgtg aagccctttt
     2641 ttcccaagat ggctggggct gtcagcatgt taaccaggat actaattggg aagttcccag
     2701 ttcgccagaa ccagccaata aggatgcacc cggtccacca atgtggaagc caagcattaa
     2761 caatggtact gatctttggg agtccaattt gagaaacgga ggtcagccgg ccgcacagca
     2821 agttccaaag ccgtcgtggg gtcatacacc atcctctaac ttaggtggaa catggggtga
     2881 ggacgacgat ggcgccgata gtagtagtgt gtggactgga ggagctgtta gcaacgcggg
     2941 atccggagct gcagtgggag taaaccaagc cggagttaat gtcggtccag gcggtgttgt
     3001 ttcgtctggc ggacctcagt ggggacaagg tgtcgttggc gtcggacttg gatcaactgg
     3061 aggtaacggg tcaagcaata taactggatc gtctggagtc gcaacaggta gtagcggaaa
     3121 ctccagcaac gctggtaacg gttggggaga ccctcgtgaa atacgccctt tgggagttgg
     3181 tggctccatg gatattcgaa atgttgaaca tcgcggcggt aacggttctg gagcaacttc
     3241 gagcgatcca cgagacattc gcatgatcga tccgcgtgac cctattcgag gagatccccg
     3301 tggaatatct ggtcgtctta atgggacctc tgaaatgtgg ggtcatcatc cacaaatgtc
     3361 ccataaccag ttgcaaggta tcaacaaaat ggttggtcaa agtgtagcaa ctgccagcac
     3421 cagtgtcgga acatctggct cgggcatcgg tcctggaggt cccggtccta gtacagtatc
     3481 aggcaatatc ccaacacagt gggggcctgc tcaaccggta agcgttggtg taagtggtcc
     3541 caaagacatg tcaaaacaga taagtggatg ggaggaacca tcaccaccgc ctcagcgtcg
     3601 cagtattcct aactacgatg atggtacatc gttgtggggt cagcaaactc gtgttcccgc
     3661 tgcaagcggt cactggaaag acatgactga ttcgataggt cgtagtagtc atctcatgcg
     3721 tggccaaagc caaacgggag gtataggaat agccggcgtt ggaaatagca atgttccagt
     3781 gggagccaat ccaagtaatc ctataagcag tgtagttgga cctcaagccc ggattccatc
     3841 tgtgggcggc gtacaacaca aaccagacgg cggcgctatg tgggtgcatt ccggcaatgt
     3901 aggtggcaga aataatgttg ctgctgttac tacttgggga gatgacactc atagcgttaa
     3961 tgtcggcgct cccagcagtg gcagtgtatc cagcaacaat tgggttgatg acaagtccaa
     4021 ctcaaccttg gcacaaaact cttggagcga cccggcccct gttggagtta gttggggcaa
     4081 taagcaaagc aaaccgccaa gcaatagtgc ttcatcaggt tggagcactg ctgcgggcgt
     4141 ggtggatggg gttgatctag gatctgagtg gaacacgcac ggggggatta ttggaaaatc
     4201 tcagcaacaa caaaaactag cgggacttaa cgtgggaatg gtgaacgtaa ttaacgcgga
     4261 gatcattaag caaagcaagc aatacaggat ccttgtcgag aacggcttta aaaaggaaga
     4321 tgtagagcgg gcattagtga ttgctaatat gaacatcgaa gaggcagccg atatgctccg
     4381 tgccaactca tccctatcaa tggatggttg gcgtcgacat gatgagtccc ttggatctta
     4441 tgccgaccac aatagttcaa caagcagcgg tggatttgct ggtcgttacc cggtcaacag
     4501 tggacaacct tcaatgtcct ttcctcataa taaccttatg aataacatgg gaggtaccgc
     4561 tgttactgga ggtaacaaca atacaaacat gacagcttta caggtgcaaa agtatttaaa
     4621 tcaagggcaa catggtgtcg ctgttggacc gcaagccgtt ggtaattctt cagcagtatc
     4681 tgtcggattt ggtcagaaca cgtctaacgc agcagtggca ggagcagcct ctgtaaatat
     4741 agcagcaaat acaaacaacc aaccgtctgg tcagcaaatt cgcatgctag gccagcaaat
     4801 tcagttggcc attcatagtg gtttcatatc tagtcagata ttgactcaac cgctaactca
     4861 aacaaccctt aaccttttaa accaacttct tagcaatatt aagcatctcc aggctgcgca
     4921 gcaatccctt acccgcgggg gaaatgtcaa tccaatggca gtgaatgtgg ctatatctaa
     4981 atacaagcag caaatccaga atttacagaa ccagataaat gcacaacagg ctgtgtatgt
     5041 aaaacagcaa aatatgcaac caacttcaca acaacaacag ccccaacaac agcaacttcc
     5101 ttctgttcat ctaagtaact caggcaacga ctatttaaga ggtcacgatg caataaataa
     5161 tttgcaaagc aacttttctg agctcaatat taataagcca agtggatatc aaggagcgtc
     5221 caatcaacaa tcccgattaa atcagtggaa gcttccagta ttagataagg agatcaactc
     5281 tgacagtacg gaattttctc gtgccccagg tgcaacgaaa caaaatttga cggccaacac
     5341 aagcaacata aactctttgg gtcttcaaaa cgatagtaca tggtcaactg gacgcagtat
     5401 tggtgacggt tggcctgatc cctcatctga taacgagaat aaagactggt ctgttgctca
     5461 gccaacttca gcagcaactg cttacactga tctggtccaa gagtttgagc caggcaagcc
     5521 atggaagggt tcacagatca aaagcataga agatgatccc agcattacac caggaagcgt
     5581 tgctagatct ccattgtcta ttaattcgac gccaaaagat gctgacatat ttgccaatac
     5641 cggtaaaaat tcaccgactg atttaccgcc actaagttta tcgtcgtcta catggagttt
     5701 taatccaaac caaaattatc cgagtcacag ttggtctgac aatagtcaac aatgtaccgc
     5761 cacttcggag ctttggacaa gcccgctaaa taaatcatcg tctcgaggtc ccccgccagg
     5821 attgactgcc aattcaaata aatctgcaaa tagtaatgcg tcaacgccaa caactattac
     5881 cggaggtgcg aatggatggt tacagcctcg aagtggcggt gttcaaacca caaacactaa
     5941 ttggacaggt ggtaacacca cttggggctc cagttggttg cttttgaaaa atctaacagc
     6001 acagattgat ggtcctactt tgcgtacact gtgtatgcag catgggcccc ttgtcagctt
     6061 tcacccgtat ttgaaccaag gaattgcctt atgtaaatat actactcgtg aggaggcgaa
     6121 caaggcgcaa atggcgttaa acaactgtgt cctcgccaac accacaatat ttgctgaatc
     6181 tcccagcgag aacgaggtgc aaagcattat gcagcactta ccacaaactc cttcctctac
     6241 aagctctagt ggaactagtg gtggcaacgt cggaggcgtc ggcacttcag ccaataatgc
     6301 aaacagtggt tctgcagctt gtctgtccgg aaacaatagc ggcaacggaa acggcagcgc
     6361 gagcggcgcc ggcagcggca acaatggcaa cagtagctgc aacaacagtg ccgccggggg
     6421 gggcagcagc agcaacaaca cgattaccac tgtagcaaat tcgaatcttg ttggttctag
     6481 tggctctgtc tcaaattcct ctggcgttac tgctaactct agtactgttt ctgtagttag
     6541 ttgtacagcg agtgggaatt ccataaatgg ggcaggtact gcaaacagtt ctggttcaaa
     6601 gagtagtgca aacaatttag ctagcggcca gtctagcgct tctaacttaa ctaatagcac
     6661 caattcaaca tggcgacaaa ctagccaaaa ccaagctctt caaagtcaaa gcaggccatc
     6721 aggcagagaa gctgactttg attatatatc tctcgtttat tccattgttg atgattaagc
     6781 ggccgctcga gtctagaggg cccttcgaag gtaagcctat ccctaaccct ctcctcggtc
     6841 tcgattctac gcgtaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag
     6901 cctcgactgt gccttctaag gcctgagctc gctgatcagc ctcgatcgag gatccagaca
     6961 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     7021 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     7081 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     7141 ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat cagtcgaccg
     7201 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc
     7261 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
     7321 ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg
     7381 cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc
     7441 aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac
     7501 gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct
     7561 tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac
     7621 catcagggac agcttcaagg atcgctcgcg gctcttacca gccagcaaaa ggccaggaac
     7681 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7741 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7801 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7861 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     7921 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7981 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     8041 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     8101 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     8161 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     8221 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     8281 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     8341 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     8401 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     8461 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     8521 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8581 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8641 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8701 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8761 cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8821 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8881 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8941 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     9001 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     9061 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     9121 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     9181 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     9241 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     9301 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     9361 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     9421 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     9481 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgt
//
