LOCUS       sequence-180799-        9619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for expression of NHA-tagged Drosophila melanogaster GW182 
            with mutated PAM2 motif in D. melanogaster cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9619)
  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 9619)
  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-695
FEATURES             Location/Qualifiers
     source          1..9619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2568
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             2589..2654
                     /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             2682..2708
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6904..6945
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6955..6972
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6995..7012)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7045..7179
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7069..7088)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7123..7142
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7801..8389)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7881..7900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8560..9351,9352..9420))
                     /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     9183..9202
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9421..9525)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9593..9611
                     /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 cggatcgggg
     2581 tacccaacat ggacgcacaa acacgacgac gtgagcgtcg cgctgagaaa caagctcaat
     2641 ggaaagctgc aaacggtgga tctcctccaa gcttgatggc ttacccatac gatgttccag
     2701 attacgctcc tgcaggcatg cgtgaagccc ttttttccca agatggctgg ggctgtcagc
     2761 atgttaacca ggatactaat tgggaagttc ccagttcgcc agaaccagcc aataaggatg
     2821 cacccggtcc accaatgtgg aagccaagca ttaacaatgg tactgatctt tgggagtcca
     2881 atttgagaaa cggaggtcag ccggccgcac agcaagttcc aaagccgtcg tggggtcata
     2941 caccatcctc taacttaggt ggaacatggg gtgaggacga cgatggcgcc gatagtagta
     3001 gtgtgtggac tggaggagct gttagcaacg cgggatccgg agctgcagtg ggagtaaacc
     3061 aagccggagt taatgtcggt ccaggcggtg ttgtttcgtc tggcggacct cagtggggac
     3121 aaggtgtcgt tggcgtcgga cttggatcaa ctggaggtaa cgggtcaagc aatataactg
     3181 gatcgtctgg agtcgcaaca ggtagtagcg gaaactccag caacgctggt aacggttggg
     3241 gagaccctcg tgaaatacgc cctttgggag ttggtggctc catggatatt cgaaatgttg
     3301 aacatcgcgg cggtaacggt tctggagcaa cttcgagcga tccacgagac attcgcatga
     3361 tcgatccgcg tgaccctatt cgaggagatc cccgtggaat atctggtcgt cttaatggga
     3421 cctctgaaat gtggggtcat catccacaaa tgtcccataa ccagttgcaa ggtatcaaca
     3481 aaatggttgg tcaaagtgta gcaactgcca gcaccagtgt cggaacatct ggctcgggca
     3541 tcggtcctgg aggtcccggt cctagtacag tatcaggcaa tatcccaaca cagtgggggc
     3601 ctgctcaacc ggtaagcgtt ggtgtaagtg gtcccaaaga catgtcaaaa cagataagtg
     3661 gatgggagga accatcacca ccgcctcagc gtcgcagtat tcctaactac gatgatggta
     3721 catcgttgtg gggtcagcaa actcgtgttc ccgctgcaag cggtcactgg aaagacatga
     3781 ctgattcgat aggtcgtagt agtcatctca tgcgtggcca aagccaaacg ggaggtatag
     3841 gaatagccgg cgttggaaat agcaatgttc cagtgggagc caatccaagt aatcctataa
     3901 gcagtgtagt tggacctcaa gcccggattc catctgtggg cggcgtacaa cacaaaccag
     3961 acggcggcgc tatgtgggtg cattccggca atgtaggtgg cagaaataat gttgctgctg
     4021 ttactacttg gggagatgac actcatagcg ttaatgtcgg cgctcccagc agtggcagtg
     4081 tatccagcaa caattgggtt gatgacaagt ccaactcaac cttggcacaa aactcttgga
     4141 gcgacccggc ccctgttgga gttagttggg gcaataagca aagcaaaccg ccaagcaata
     4201 gtgcttcatc aggttggagc actgctgcgg gcgtggtgga tggggttgat ctaggatctg
     4261 agtggaacac gcacgggggg attattggaa aatctcagca acaacaaaaa ctagcgggac
     4321 ttaacgtggg aatggtgaac gtaattaacg cggagatcat taagcaaagc aagcaataca
     4381 ggatccttgt cgagaacggc tttaaaaagg aagatgtaga gcgggcatta gtgattgcta
     4441 atatgaacat cgaagaggca gccgatatgc tccgtgccaa ctcatcccta tcaatggatg
     4501 gttggcgtcg acatgatgag tcccttggat cttatgccga ccacaatagt tcaacaagca
     4561 gcggtggatt tgctggtcgt tacccggtca acagtggaca accttcaatg tcctttcctc
     4621 ataataacct tatgaataac atgggaggta ccgctgttac tggaggtaac aacaatacaa
     4681 acatgacagc tttacaggtg caaaagtatt taaatcaagg gcaacatggt gtcgctgttg
     4741 gaccgcaagc cgttggtaat tcttcagcag tatctgtcgg atttggtcag aacacgtcta
     4801 acgcagcagt ggcaggagca gcctctgtaa atatagcagc aaatacaaac aaccaaccgt
     4861 ctggtcagca aattcgcatg ctaggccagc aaattcagtt ggccattcat agtggtttca
     4921 tatctagtca gatattgact caaccgctaa ctcaaacaac ccttaacctt ttaaaccaac
     4981 ttcttagcaa tattaagcat ctccaggctg cgcagcaatc ccttacccgc gggggaaatg
     5041 tcaatccaat ggcagtgaat gtggctatat ctaaatacaa gcagcaaatc cagaatttac
     5101 agaaccagat aaatgcacaa caggctgtgt atgtaaaaca gcaaaatatg caaccaactt
     5161 cacaacaaca acagccccaa caacagcaac ttccttctgt tcatctaagt aactcaggca
     5221 acgactattt aagaggtcac gatgcaataa ataatttgca aagcaacttt tctgagctca
     5281 atattaataa gccaagtgga tatcaaggag cgtccaatca acaatcccga ttaaatcagt
     5341 ggaagcttcc agtattagat aaggagatca actctgacag tacggaattt tctcgtgccc
     5401 caggtgcaac gaaacaaaat ttgacggcca acacaagcaa cataaactct ttgggtcttc
     5461 aaaacgatag tacatggtca actggacgca gtattggtga cggttggcct gatccctcat
     5521 ctgataacga gaataaagac tggtctgttg ctcagccaac ttcagcagca actgcttaca
     5581 ctgatctggt ccaagcggcg gagccaggca agccagcggc gggttcacag atcaaaagca
     5641 tagaagatga tcccagcatt gcgccaggaa gcgttgctag atctccattg tctattaatt
     5701 cgacgccaaa agatgctgac atatttgcca ataccggtaa aaattcaccg actgatttac
     5761 cgccactaag tttatcgtcg tctacatgga gttttaatcc aaaccaaaat tatccgagtc
     5821 acagttggtc tgacaatagt caacaatgta ccgccacttc ggagctttgg acaagcccgc
     5881 taaataaatc atcgtctcga ggtcccccgc caggattgac tgccaattca aataaatctg
     5941 caaatagtaa tgcgtcaacg ccaacaacta ttaccggagg tgcgaatgga tggttacagc
     6001 ctcgaagtgg cggtgttcaa accacaaaca ctaattggac aggtggtaac accacttggg
     6061 gctccagttg gttgcttttg aaaaatctaa cagcacagat tgatggtcct actttgcgta
     6121 cactgtgtat gcagcatggg ccccttgtca gctttcaccc gtatttgaac caaggaattg
     6181 ccttatgtaa atatactact cgtgaggagg cgaacaaggc gcaaatggcg ttaaacaact
     6241 gtgtcctcgc caacaccaca atatttgctg aatctcccag cgagaacgag gtgcaaagca
     6301 ttatgcagca cttaccacaa actccttcct ctacaagctc tagtggaact agtggtggca
     6361 acgtcggagg cgtcggcact tcagccaata atgcaaacag tggttctgca gcttgtctgt
     6421 ccggaaacaa tagcggcaac ggaaacggca gcgcgagcgg cgccggcagc ggcaacaatg
     6481 gcaacagtag ctgcaacaac agtgccgccg gggggggcag cagcagcaac aacacgatta
     6541 ccactgtagc aaattcgaat cttgttggtt ctagtggctc tgtctcaaat tcctctggcg
     6601 ttactgctaa ctctagtact gtttctgtag ttagttgtac agcgagtggg aattccataa
     6661 atggggcagg tactgcaaac agttctggtt caaagagtag tgcaaacaat ttagctagcg
     6721 gccagtctag cgcttctaac ttaactaata gcaccaattc aacatggcga caaactagcc
     6781 aaaaccaagc tcttcaaagt caaagcaggc catcaggcag agaagctgac tttgattata
     6841 tatctctcgt ttattccatt gttgatgatt aagcggccgc tcgagtctag agggcccttc
     6901 gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat
     6961 caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc taaggcctga
     7021 gctcgctgat cagcctcgat cgaggatcca gacatgataa gatacattga tgagtttgga
     7081 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     7141 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     7201 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     7261 aaatgtggta tggctgatta tgatcagtcg accgatgccc ttgagagcct tcaacccagt
     7321 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
     7381 tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg
     7441 ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc
     7501 cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat
     7561 tatcgccggc atggcggccg acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg
     7621 ctggatggcc ttccccatta tgattcttct cgcttccggc ggcatcggga tgcccgcgtt
     7681 gcaggccatg ctgtccaggc aggtagatga cgaccatcag ggacagcttc aaggatcgct
     7741 cgcggctctt accagccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7801 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7861 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7921 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7981 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     8041 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     8101 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     8161 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     8221 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     8281 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     8341 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     8401 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     8461 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     8521 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     8581 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     8641 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     8701 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     8761 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8821 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg
     8881 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8941 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     9001 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     9061 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     9121 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg
     9181 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     9241 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     9301 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     9361 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     9421 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     9481 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     9541 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     9601 tatcacgagg ccctttcgt
//
