LOCUS       sequence-194408-        6305 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for expression of NHA-tagged Drosophila melanogaster GW182 
            NED in D. melanogaster cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6305)
  AUTHORS   Chekulaeva M, Parker R, Filipowicz W
  TITLE     The GW/WG repeats of Drosophila GW182 function as effector motifs 
            for miRNA-mediated repression.
  JOURNAL   Nucleic Acids Res. 2010 Oct;38(19):6673-83. doi: 10.1093/nar/gkq501.
            Epub 2010 Jun 8.
  PUBMED    20530530
REFERENCE   2  (bases 1 to 6305)
  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: 
            "10.1093/nar/gkq501"; journalName: "Nucleic Acids Res"; date: 
            "2010-10"; volume: "38"; issue: "19"; pages: "6673-83"
COMMENT     Sequence Label: pMCh-139
FEATURES             Location/Qualifiers
     source          1..6305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(988..1006)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1074..1178
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1179..1247,1248..2039)
                     /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     complement(1397..1416)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2210..2798
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2699..2718
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3399..3533
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3436..3455)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3490..3509
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3566..3583
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3606..3623)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3633..3674)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(4580..4606)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(4634..4699)
                     /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"
     promoter        complement(join(4720..6305,1..924))
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
ORIGIN
        1 gccacagacc ccgccacgcc ccgccccacc cttccttccg ctgccccgca cccctttcct
       61 tccactaccg ttttgccgca tttggtattg ccatctcgct tttttggctt tactttgctt
      121 tttgaataag ccaagaaact gaaatcaaag cggacagtcc gaaaattgaa tattccaaaa
      181 aacaacaatt gcacatgcac tgcgttcggc aaaagacgca gaaattcaag ccttttggtc
      241 tggcatcatc ttaaaatatc atcagtatca tcattgccat tataaataca taagaaaatg
      301 agaatacatt aaaggtatat acatacaatc tttgtcatag tcaacatatt ttgcaacaaa
      361 attatctata gataacatgt ttttgtagaa aattccacta actgattttc gtataaatac
      421 aaattattct cattcttcaa aatactcgcc gacgtcatcg cgaaattcct cagcattctt
      481 cgacaaatga agagagacag cgagtgacac aatcttatct tgccgacgag tctttgaaaa
      541 tggccccggg taacggtgaa aaacatatag ccaagaatgg agcgatcgcc aggcgatgga
      601 aattggcttc aaattgctcc gaatatggcc agttggtcag ttggcattgg attgtggaga
      661 gatactagat cagctcgttc tccttctcct ccgcattctc ctactccgcc cgaagacgct
      721 tctttgatcg cttctttctc gaatgtgtac atattcatta tgtatttggt gtttggcgac
      781 gatcagccag ctttaatggc cataaaacgt gtcttggttt gtgggccaca taacaggcag
      841 cgagatctgg aaaaatgatg tgacagtgga aatgagaagt aggtgcatct gcaaagattt
      901 gattcagagt tgatgccatt catgattttt ttttagaagt atcatttgtg tttttagaat
      961 atagaattaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     1021 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     1081 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1141 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     1201 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     1261 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     1321 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     1381 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     1441 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     1501 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     1561 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     1621 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     1681 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt
     1741 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     1801 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     1861 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     1921 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     1981 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     2041 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     2101 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     2161 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2221 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2281 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2341 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2401 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2461 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2521 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2581 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2641 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2701 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2761 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2821 tacggttcct ggccttttgc tggccttgaa gctgtccctg atggtcgtca tctacctgcc
     2881 tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa
     2941 tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg
     3001 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     3061 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     3121 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     3181 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     3241 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga ctgatcataa
     3301 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3361 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3421 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3481 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcctcgatc
     3541 gaggctgatc agcgagctca ggccttagaa ggcacagtcg aggctgatca gcgggtttaa
     3601 actcaatggt gatggtgatg atgaccggta cgcgtagaat cgagaccgag gagagggtta
     3661 gggataggct taccttcgaa ccgcgggccc tctagactcg agcggccgct tatttgcttt
     3721 gcttattgcc ccaactaact ccaacagggg ccgggtcgct ccaagagttt tgtgccaagg
     3781 ttgagttgga cttgtcatca acccaattgt tgctggatac actgccactg ctgggagcgc
     3841 cgacattaac gctatgagtg tcatctcccc aagtagtaac agcagcaaca ttatttctgc
     3901 cacctacatt gccggaatgc acccacatag cgccgccgtc tggtttgtgt tgtacgccgc
     3961 ccacagatgg aatccgggct tgaggtccaa ctacactgct tataggatta cttggattgg
     4021 ctcccactgg aacattgcta tttccaacgc cggctattcc tatacctccc gtttggcttt
     4081 ggccacgcat gagatgacta ctacgaccta tcgaatcagt catgtctttc cagtgaccgc
     4141 ttgcagcggg aacacgagtt tgctgacccc acaacgatgt accatcatcg tagttaggaa
     4201 tactgcgacg ctgaggcggt ggtgatggtt cctcccatcc acttatctgt tttgacatgt
     4261 ctttgggacc acttacacca acgcttaccg gttgagcagg cccccactgt gttgggatat
     4321 tgcctgatac tgtactagga ccgggacctc caggaccgat gcccgagcca gatgttccga
     4381 cactggtgct ggcagttgct acactttgac caaccatttt gttgatacct tgcaactggt
     4441 tatgggacat ttgtggatga tgaccccaca tttcagaggt cccattaaga cgaccagata
     4501 ttccacgggg atctcctcga atagggtcac gcggatcgat catgcgaatg tctcgtggat
     4561 cgctcgaagt gcctgcagga gcgtaatctg gaacatcgta tgggtaagcc atcaagcttg
     4621 gaggagatcc accgtttgca gctttccatt gagcttgttt ctcagcgcga cgctcacgtc
     4681 gtcgtgtttg tgcgtccatg ttgggtaccc cgatccgggg tctctggatt agacgactgc
     4741 tggctgatgg agcggctttg tgtcgggagg agtatccaca cagcacaaga actcaaacgg
     4801 tagtgatatg aaaactggtc ccgcagcccg ttttataaag cccgcttagc gctaaaacct
     4861 tatcagccgc aatgctctcc gctctcaagt cgcgttcaaa acttttacca tatatgggta
     4921 tgaattgttc gccgattggg ttttcattta cggagaattt cctccgcaac tgggtgttcg
     4981 tgcttgtgtg tgtcttgtat atcgccatga agagtgtact tccatgggtc ctatcagttg
     5041 cggcatgtga ttcaatttgg gggttactca tattcgatac ttcagctggt tatctttatg
     5101 ctgtgatggt cacggtgctt ttacacattt tttccccaac tactcattgt atgccaaaac
     5161 atacatatgt acatgtgttt atagtgggtt tattagtaat gtccagcgat attaaaacat
     5221 tacagttaaa taatattcat attagcttac gtttactgct atctgcactt caaattggtt
     5281 attccaggga cattttgtgc aacacacatc ttctagtagg gcattcacta gtcgtagaaa
     5341 acacacctaa cttaagccgg ttttaacaat aacggcgaaa cagctgccag acgagagaaa
     5401 agtacacagc aagaagcatt tctataatga tgccgtccta aatatgcagc atttttgctc
     5461 aaattgatca tttcgttgca tcgctcaggt ggtatagtat ttggctattg aatgtgattg
     5521 ctatcttaat agttttgaat agtaatagtg caaatattca ttcataaaaa ataggtaatc
     5581 gattttttca gtgcagtcta tgcgtttgcg caacagagac gaagagagag acgcctcaca
     5641 cacaagcaca tgcggtctaa gctgctctcc ctctttttct ctctctctct ggcttgcggc
     5701 agttttccag aggctgtgac tcagtaagca tttcttgttg ttaccctaac gataagcgtc
     5761 cgattcctaa ttctctttca aataatttgt tttatttttt tttttgtgtg ttttgaagtc
     5821 aaacagcttc ataaacaata ttaaatatcg caattgttat tgcaaatgca ttcgtatcag
     5881 gcgccttgat gtttttcgcc aaaacttcgt ctgctctttg tcagcgtaat tgcgtttgcc
     5941 ggtcgtgccg tcttgctcgc acgcggcttg caaacaaaaa gtgaaagagg gaaagccacc
     6001 tgtgttgttg cttttgttgc tatcgttggc tgcacacaca tggcaataag taaccatatt
     6061 gtagacttat ttattttagc gttttcgttg actaatcttt gtttaaataa ttgcctattt
     6121 cgagccaaaa tgacgcataa aaggaaacgg caacatgcgg tgcgttgagc atgcgtaagc
     6181 gagatggcgc tagagagagc gcagcataaa aagaaaaaaa caaccggcga gtggcgagac
     6241 acgaataata cacacacgca cacatggtca aaggagtggc tccaacgtcg gtgacgtcgc
     6301 gccca
//
