LOCUS       sequence-194511-        9509 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 9509)
  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 9509)
  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..9509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1847..1873
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6073..6114
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6124..6141
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6164..6181)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6214..6348
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6238..6257)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6292..6311
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6949..7537)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7029..7048)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7708..8499,8500..8568))
                     /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     8331..8350
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8569..8673)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8741..8759
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(8823..9509,1..1823)
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
ORIGIN
        1 caaaaggctt gaatttctgc gtcttttgcc gaacgcagtg catgtgcaat tgttgttttt
       61 tggaatattc aattttcgga ctgtccgctt tgatttcagt ttcttggctt attcaaaaag
      121 caaagtaaag ccaaaaaagc gagatggcaa taccaaatgc ggcaaaacgg tagtggaagg
      181 aaaggggtgc ggggcagcgg aaggaagggt ggggcggggc gtggcggggt ctgtggctgg
      241 gcgcgacgtc accgacgttg gagccactcc tttgaccatg tgtgcgtgtg tgtattattc
      301 gtgtctcgcc actcgccggt tgtttttttc tttttatgct gcgctctctc tagcgccatc
      361 tcgcttacgc atgctcaacg caccgcatgt tgccgtttcc ttttatgcgt cattttggct
      421 cgaaataggc aattatttaa acaaagatta gtcaacgaaa acgctaaaat aaataagtct
      481 acaatatggt tacttattgc catgtgtgtg cagccaacga tagcaacaaa agcaacaaca
      541 caggtggctt tccctctttc actttttgtt tgcaagccgc gtgcgagcaa gacggcacga
      601 ccggcaaacg caattacgct gacaaagagc agacgaagtt ttggcgaaaa acatcaaggc
      661 gcctgatacg aatgcatttg caataacaat tgcgatattt aatattgttt atgaagctgt
      721 ttgacttcaa aacacacaaa aaaaaaaata aaacaaatta tttgaaagag aattaggaat
      781 cggacgctta tcgttagggt aacaacaaga aatgcttact gagtcacagc ctctggaaaa
      841 ctgccgcaag ccagagagag agagaaaaag agggagagca gcttagaccg catgtgcttg
      901 tgtgtgaggc gtctctctct tcgtctctgt tgcgcaaacg catagactgc actgaaaaaa
      961 tcgattacct attttttatg aatgaatatt tgcactatta ctattcaaaa ctattaagat
     1021 agcaatcaca ttcaatagcc aaatactata ccacctgagc gatgcaacga aatgatcaat
     1081 ttgagcaaaa atgctgcata tttaggacgg catcattata gaaatgcttc ttgctgtgta
     1141 cttttctctc gtctggcagc tgtttcgccg ttattgttaa aaccggctta agttaggtgt
     1201 gttttctacg actagtgaat gccctactag aagatgtgtg ttgcacaaaa tgtccctgga
     1261 ataaccaatt tgaagtgcag atagcagtaa acgtaagcta atatgaatat tatttaactg
     1321 taatgtttta atatcgctgg acattactaa taaacccact ataaacacat gtacatatgt
     1381 atgttttggc atacaatgag tagttgggga aaaaatgtgt aaaagcaccg tgaccatcac
     1441 agcataaaga taaccagctg aagtatcgaa tatgagtaac ccccaaattg aatcacatgc
     1501 cgcaactgat aggacccatg gaagtacact cttcatggcg atatacaaga cacacacaag
     1561 cacgaacacc cagttgcgga ggaaattctc cgtaaatgaa aacccaatcg gcgaacaatt
     1621 catacccata tatggtaaaa gttttgaacg cgacttgaga gcggagagca ttgcggctga
     1681 taaggtttta gcgctaagcg ggctttataa aacgggctgc gggaccagtt ttcatatcac
     1741 taccgtttga gttcttgtgc tgtgtggata ctcctcccga cacaaagccg ctccatcagc
     1801 cagcagtcgt ctaatccaga gaccccggat cgggagcttg atggcttacc catacgatgt
     1861 tccagattac gctcctgcag gcatgcgtga agcccttttt tcccaagatg gctggggctg
     1921 tcagcatgtt aaccaggata ctaattggga agttcccagt tcgccagaac cagccaataa
     1981 ggatgcaccc ggtccaccaa tgtggaagcc aagcattaac aatggtactg atctttggga
     2041 gtccaatttg agaaacggag gtcagccggc cgcacagcaa gttccaaagc cgtcgtgggg
     2101 tcatacacca tcctctaact taggtggaac atggggtgag gacgacgatg gcgccgatag
     2161 tagtagtgtg tggactggag gagctgttag caacgcggga tccggagctg cagtgggagt
     2221 aaaccaagcc ggagttaatg tcggtccagg cggtgttgtt tcgtctggcg gacctcagtg
     2281 gggacaaggt gtcgttggcg tcggacttgg atcaactgga ggtaacgggt caagcaatat
     2341 aactggatcg tctggagtcg caacaggtag tagcggaaac tccagcaacg ctggtaacgg
     2401 ttggggagac cctcgtgaaa tacgcccttt gggagttggt ggctccatgg atattcgaaa
     2461 tgttgaacat cgcggcggta acggttctgg agcaacttcg agcgatccac gagacattcg
     2521 catgatcgat ccgcgtgacc ctattcgagg agatccccgt ggaatatctg gtcgtcttaa
     2581 tgggacctct gaaatgtggg gtcatcatcc acaaatgtcc cataaccagt tgcaaggtat
     2641 caacaaaatg gttggtcaaa gtgtagcaac tgccagcacc agtgtcggaa catctggctc
     2701 gggcatcggt cctggaggtc ccggtcctag tacagtatca ggcaatatcc caacacagtg
     2761 ggggcctgct caaccggtaa gcgttggtgt aagtggtccc aaagacatgt caaaacagat
     2821 aagtggatgg gaggaaccat caccaccgcc tcagcgtcgc agtattccta actacgatga
     2881 tggtacatcg ttgtggggtc agcaaactcg tgttcccgct gcaagcggtc actggaaaga
     2941 catgactgat tcgataggtc gtagtagtca tctcatgcgt ggccaaagcc aaacgggagg
     3001 tataggaata gccggcgttg gaaatagcaa tgttccagtg ggagccaatc caagtaatcc
     3061 tataagcagt gtagttggac ctcaagcccg gattccatct gtgggcggcg tacaacacaa
     3121 accagacggc ggcgctatgt gggtgcattc cggcaatgta ggtggcagaa ataatgttgc
     3181 tgctgttact acttggggag atgacactca tagcgttaat gtcggcgctc ccagcagtgg
     3241 cagtgtatcc agcaacaatt gggttgatga caagtccaac tcaaccttgg cacaaaactc
     3301 ttggagcgac ccggcccctg ttggagttag ttggggcaat aagcaaagca aaccgccaag
     3361 caatagtgct tcatcaggtt ggagcactgc tgcgggcgtg gtggatgggg ttgatctagg
     3421 atctgagtgg aacacgcacg gggggattat tggaaaatct cagcaacaac aaaaactagc
     3481 gggacttaac gtgggaatgg tgaacgtaat taacgcggag atcattaagc aaagcaagca
     3541 atacaggatc cttgtcgaga acggctttaa aaaggaagat gtagagcggg cattagtgat
     3601 tgctaatatg aacatcgaag aggcagccga tatgctccgt gccaactcat ccctatcaat
     3661 ggatggttgg cgtcgacatg atgagtccct tggatcttat gccgaccaca atagttcaac
     3721 aagcagcggt ggatttgctg gtcgttaccc ggtcaacagt ggacaacctt caatgtcctt
     3781 tcctcataat aaccttatga ataacatggg aggtaccgct gttactggag gtaacaacaa
     3841 tacaaacatg acagctttac aggtgcaaaa gtatttaaat caagggcaac atggtgtcgc
     3901 tgttggaccg caagccgttg gtaattcttc agcagtatct gtcggatttg gtcagaacac
     3961 gtctaacgca gcagtggcag gagcagcctc tgcaaatata gcagcaaata caaacaacca
     4021 accgtctggt cagcaaattc gcatgctagg ccagcaaatt cagttggcca ttcatagtgg
     4081 tttcatatct agtcagatat tgactcaacc gctaactcaa acaaccctta accttttaaa
     4141 ccaacttctt agcaatatta agcatctcca ggctgcacag caatccctta cccgcggggg
     4201 aaatgtcaat ccaatggcag tgaatgtggc tatatctaaa tacaagcagc aaatccagaa
     4261 tttacagaac cagataagtg cacaacaggc tgtgtatgta aaacagcaaa atatgcaacc
     4321 aacttcacaa caacaacagc cccaacaaca gcaacttcct tctgttcatc taagtaactc
     4381 aggcaacgac tatttaagag gtcacgatgc aataaataat ttgcaaagca acttttctga
     4441 gctcaatatt aataagccaa gtggatatca aggagcgtcc aatcaacaat cccgattaaa
     4501 tcagtggaag cttccagtat tagataagga gatcaactct gacagtacgg aattttctcg
     4561 tgccccaggt gcaacgaaac aaaatttgac ggccaacaca agcaacataa actctttggg
     4621 tcttcaaaac gatagtacat ggtcaactgg acgcagtatt ggtgacggtt ggcctgatcc
     4681 ctcatctgat aacgagaata aagactggtc tgttgctcag ccaacttcag cagcaactgc
     4741 ttacactgat ctggtccaag agtttgagcc aggcaagcca tggaagggtt cacagatcaa
     4801 aagcatagaa gatgatccca gcattacacc aggaagcgtt gctagatctc cattgtctat
     4861 taattcgacg ccaaaagatg ctgacatatt tgccaatacc ggtaaaaatt caccgactga
     4921 tttaccgcca ctaagtttat cgtcgtctac atggagtttt aatccaaacc aaaattatcc
     4981 gagtcacagt tggtctgaca atagtcaaca atgtaccgcc acttcggagc tttggacaag
     5041 cccgctaaat aaatcatcgt ctcgaggtcc cccgccagga ttgactgcca attcaaataa
     5101 atctgcaaat agtaatgcgt caacgccaac aactattacc ggaggtgcga atggatggtt
     5161 acagcctcga agtggcggtg ttcaaaccac aaacactaat tggacaggtg gtaacaccac
     5221 ttggggctcc agttggttgc ttttgaaaaa tctaacagca cagattgatg gtcctacttt
     5281 gcgtacactg tgtatgcagc atgggcccct tgtcagcttt cacccgtatt tgaaccaagg
     5341 aattgcctta tgtaaatata ctactcgtga ggaggcgaac aaggcgcaaa tggcgttaaa
     5401 caactgtgtc ctcgccaaca ccacaatatt tgctgaatct cccagcgaga acgaggtgca
     5461 aagcattatg cagcacttac cacaaactcc ttcctctaca agctctagtg gaactagtgg
     5521 tggcaacgtc ggaggcgtcg gcacttcagc caataatgca aacagtggtt ctgcagcttg
     5581 tctgtccgga aacaatagcg gcaacggaaa cggcagcgcg agcggcgccg gcagcggcaa
     5641 caatggcaac agtagctgca acaacagtgc cgccgggggg ggcagcagca gcaacaacac
     5701 gattaccact gtagcaaatt cgaatcttgt tggttctagt ggctctgtct caaattcctc
     5761 tggcgttact gctaactcta gtactgtttc tgtagttagt tgtacagcga gtgggaattc
     5821 cataaatggg gcaggtactg caaacagttc tggttcaaag agtagtgcaa acaatttagc
     5881 tagcggccag tctagcgctt ctaacttaac taatagcacc aattcaacat ggcgacaaac
     5941 tagccaaaac caagctcttc aaagtcaaag caggccatca ggcagagaag ctgactttga
     6001 ttatatatct ctcgtttatt ccattgttga tgattaagcg gccgctcgag tctagagggc
     6061 ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc tacgcgtacc
     6121 ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac tgtgccttct
     6181 aaggcctgag ctcgctgatc agcctcgatc gaggatccag acatgataag atacattgat
     6241 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     6301 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     6361 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     6421 aacctctaca aatgtggtat ggctgattat gatcagtcga ccgatgccct tgagagcctt
     6481 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     6541 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg
     6601 cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat
     6661 cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa
     6721 gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc
     6781 gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg gcatcgggat
     6841 gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg gacagcttca
     6901 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6961 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7021 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7081 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7141 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7201 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7261 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7321 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7381 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7441 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7501 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7561 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7621 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7681 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7741 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7801 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7861 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7921 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7981 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc
     8041 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8101 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8161 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8221 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8281 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc
     8341 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8401 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8461 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8521 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8581 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8641 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8701 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     8761 ctttcgtctt caagaattaa ttctatattc taaaaacaca aatgatactt ctaaaaaaaa
     8821 atcatgaatg gcatcaactc tgaatcaaat ctttgcagat gcacctactt ctcatttcca
     8881 ctgtcacatc atttttccag atctcgctgc ctgttatgtg gcccacaaac caagacacgt
     8941 tttatggcca ttaaagctgg ctgatcgtcg ccaaacacca aatacataat gaatatgtac
     9001 acattcgaga aagaagcgat caaagaagcg tcttcgggcg gagtaggaga atgcggagga
     9061 gaaggagaac gagctgatct agtatctctc cacaatccaa tgccaactga ccaactggcc
     9121 atattcggag caatttgaag ccaatttcca tcgcctggcg atcgctccat tcttggctat
     9181 atgtttttca ccgttacccg gggccatttt caaagactcg tcggcaagat aagattgtgt
     9241 cactcgctgt ctctcttcat ttgtcgaaga atgctgagga atttcgcgat gacgtcggcg
     9301 agtattttga agaatgagaa taatttgtat ttatacgaaa atcagttagt ggaattttct
     9361 acaaaaacat gttatctata gataattttg ttgcaaaata tgttgactat gacaaagatt
     9421 gtatgtatat acctttaatg tattctcatt ttcttatgta tttataatgg caatgatgat
     9481 actgatgata ttttaagatg atgccagac
//
