LOCUS       sequence-194414-        6275 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for expression of NHA-tagged Drosophila melanogaster GW182 
            CED in D. melanogaster cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6275)
  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 6275)
  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-76
FEATURES             Location/Qualifiers
     source          1..6275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(968..986)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1054..1158
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1159..1227,1228..2019)
                     /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(1377..1396)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2190..2778
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2679..2698
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3379..3513
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3416..3435)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3470..3489
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3546..3563
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3586..3603)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3613..3654)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(4530..4556)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(4584..4649)
                     /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(4670..6275,1..904))
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
ORIGIN
        1 ccgccccacc cttccttccg ctgccccgca cccctttcct tccactaccg ttttgccgca
       61 tttggtattg ccatctcgct tttttggctt tactttgctt tttgaataag ccaagaaact
      121 gaaatcaaag cggacagtcc gaaaattgaa tattccaaaa aacaacaatt gcacatgcac
      181 tgcgttcggc aaaagacgca gaaattcaag ccttttggtc tggcatcatc ttaaaatatc
      241 atcagtatca tcattgccat tataaataca taagaaaatg agaatacatt aaaggtatat
      301 acatacaatc tttgtcatag tcaacatatt ttgcaacaaa attatctata gataacatgt
      361 ttttgtagaa aattccacta actgattttc gtataaatac aaattattct cattcttcaa
      421 aatactcgcc gacgtcatcg cgaaattcct cagcattctt cgacaaatga agagagacag
      481 cgagtgacac aatcttatct tgccgacgag tctttgaaaa tggccccggg taacggtgaa
      541 aaacatatag ccaagaatgg agcgatcgcc aggcgatgga aattggcttc aaattgctcc
      601 gaatatggcc agttggtcag ttggcattgg attgtggaga gatactagat cagctcgttc
      661 tccttctcct ccgcattctc ctactccgcc cgaagacgct tctttgatcg cttctttctc
      721 gaatgtgtac atattcatta tgtatttggt gtttggcgac gatcagccag ctttaatggc
      781 cataaaacgt gtcttggttt gtgggccaca taacaggcag cgagatctgg aaaaatgatg
      841 tgacagtgga aatgagaagt aggtgcatct gcaaagattt gattcagagt tgatgccatt
      901 catgattttt ttttagaagt atcatttgtg tttttagaat atagaattaa ttcttgaaga
      961 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     1021 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1081 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1141 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1201 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1261 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1321 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1381 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     1441 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1501 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1561 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     1621 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     1681 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     1741 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     1801 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     1861 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     1921 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     1981 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2041 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2101 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2161 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2221 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2281 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     2341 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2401 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2461 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2521 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2581 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2641 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2701 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2761 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     2821 tggccttgaa gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac
     2881 gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct
     2941 cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg
     3001 gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg
     3061 tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg
     3121 tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag
     3181 aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact
     3241 gggttgaagg ctctcaaggg catcggtcga ctgatcataa tcagccatac cacatttgta
     3301 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     3361 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3421 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3481 aaactcatca atgtatctta tcatgtctgg atcctcgatc gaggctgatc agcgagctca
     3541 ggccttagaa ggcacagtcg aggctgatca gcgggtttaa actcaatggt gatggtgatg
     3601 atgaccggta cgcgtagaat cgagaccgag gagagggtta gggataggct taccttcgaa
     3661 ccgcgggccc tctagactcg agcggccgct tagttgccac cactagttcc actagagctt
     3721 gtagaggaag gagtttgtgg taagtgctgc ataatgcttt gcacctcgtt ctcgctggga
     3781 gattcagcaa atattgtggt gttggcgagg acacagttgt ttaacgccat ttgcgccttg
     3841 ttcgcctcct cacgagtagt atatttacat aaggcaattc cttggttcaa atacgggtga
     3901 aagctgacaa ggggcccatg ctgcatacac agtgtacgca aagtaggacc atcaatctgt
     3961 gctgttagat ttttcaaaag caaccaactg gagccccaag tggtgttacc acctgtccaa
     4021 ttagtgtttg tggtttgaac accgccactt cgaggctgta accatccatt cgcacctccg
     4081 gtaatagttg ttggcgttga cgcattacta tttgcagatt tatttgaatt ggcagtcaat
     4141 cctggcgggg gacctcgaga cgatgattta tttagcgggc ttgtccaaag ctccgaagtg
     4201 gcggtacatt gttgactatt gtcagaccaa ctgtgactcg gataattttg gtttggatta
     4261 aaactccatg tagacgacga taaacttagt ggcggtaaat cagtcggtga atttttaccg
     4321 gtattggcaa atatgtcagc atcttttggc gtcgaattaa tagacaatgg agatctagca
     4381 acgcttcctg gtgtaatgct gggatcatct tctatgcttt tgatctgtga acccttccat
     4441 ggcttgcctg gctcaaactc ttggaccaga tcagtgtaag cagttgctgc tgaagttggc
     4501 tgagcaacag accagtcttt gcctgcagga gcgtaatctg gaacatcgta tgggtaagcc
     4561 atcaagcttg gaggagatcc accgtttgca gctttccatt gagcttgttt ctcagcgcga
     4621 cgctcacgtc gtcgtgtttg tgcgtccatg ttgggtaccc cgatccgggg tctctggatt
     4681 agacgactgc tggctgatgg agcggctttg tgtcgggagg agtatccaca cagcacaaga
     4741 actcaaacgg tagtgatatg aaaactggtc ccgcagcccg ttttataaag cccgcttagc
     4801 gctaaaacct tatcagccgc aatgctctcc gctctcaagt cgcgttcaaa acttttacca
     4861 tatatgggta tgaattgttc gccgattggg ttttcattta cggagaattt cctccgcaac
     4921 tgggtgttcg tgcttgtgtg tgtcttgtat atcgccatga agagtgtact tccatgggtc
     4981 ctatcagttg cggcatgtga ttcaatttgg gggttactca tattcgatac ttcagctggt
     5041 tatctttatg ctgtgatggt cacggtgctt ttacacattt tttccccaac tactcattgt
     5101 atgccaaaac atacatatgt acatgtgttt atagtgggtt tattagtaat gtccagcgat
     5161 attaaaacat tacagttaaa taatattcat attagcttac gtttactgct atctgcactt
     5221 caaattggtt attccaggga cattttgtgc aacacacatc ttctagtagg gcattcacta
     5281 gtcgtagaaa acacacctaa cttaagccgg ttttaacaat aacggcgaaa cagctgccag
     5341 acgagagaaa agtacacagc aagaagcatt tctataatga tgccgtccta aatatgcagc
     5401 atttttgctc aaattgatca tttcgttgca tcgctcaggt ggtatagtat ttggctattg
     5461 aatgtgattg ctatcttaat agttttgaat agtaatagtg caaatattca ttcataaaaa
     5521 ataggtaatc gattttttca gtgcagtcta tgcgtttgcg caacagagac gaagagagag
     5581 acgcctcaca cacaagcaca tgcggtctaa gctgctctcc ctctttttct ctctctctct
     5641 ggcttgcggc agttttccag aggctgtgac tcagtaagca tttcttgttg ttaccctaac
     5701 gataagcgtc cgattcctaa ttctctttca aataatttgt tttatttttt tttttgtgtg
     5761 ttttgaagtc aaacagcttc ataaacaata ttaaatatcg caattgttat tgcaaatgca
     5821 ttcgtatcag gcgccttgat gtttttcgcc aaaacttcgt ctgctctttg tcagcgtaat
     5881 tgcgtttgcc ggtcgtgccg tcttgctcgc acgcggcttg caaacaaaaa gtgaaagagg
     5941 gaaagccacc tgtgttgttg cttttgttgc tatcgttggc tgcacacaca tggcaataag
     6001 taaccatatt gtagacttat ttattttagc gttttcgttg actaatcttt gtttaaataa
     6061 ttgcctattt cgagccaaaa tgacgcataa aaggaaacgg caacatgcgg tgcgttgagc
     6121 atgcgtaagc gagatggcgc tagagagagc gcagcataaa aagaaaaaaa caaccggcga
     6181 gtggcgagac acgaataata cacacacgca cacatggtca aaggagtggc tccaacgtcg
     6241 gtgacgtcgc gcccagccac agaccccgcc acgcc
//
