LOCUS       sequence-355870-        9590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmGW182-10xmut.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9590)
  AUTHORS   Eulalio A, Helms S, Fritzsch C, Fauser M, Izaurralde E
  TITLE     A C-terminal silencing domain in GW182 is essential for miRNA 
            function.
  JOURNAL   RNA. 2009 Jun;15(6):1067-77. doi: 10.1261/rna.1605509. Epub 2009 Apr
            21.
  PUBMED    19383769
REFERENCE   2  (bases 1 to 9590)
  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.1605509"; journalName: "RNA"; date: "2009-06"; volume: 
            "15"; issue: "6"; pages: "1067-77"
COMMENT     Sequence Label: pAc5.1B-lambdaN-HA-DmGW182-10xmut_G
FEATURES             Location/Qualifiers
     source          1..9590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..2510
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             2531..2596
                     /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             2624..2650
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6841..6882
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6892..6909
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6932..6949)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6982..7116
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7006..7025)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7060..7079
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7717..8305)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7797..7816)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8476..9267,9268..9336))
                     /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     9099..9118
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9337..9441)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9509..9527
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 catgaatggc atcaactctg aatcaaatct ttgcagatgc acctacttct catttccact
       61 gtcacatcat ttttccagat ctcgctgcct gttatgtggc ccacaaacca agacacgttt
      121 tatggccatt aaagctggct gatcgtcgcc aaacaccaaa tacataatga atatgtacac
      181 attcgagaaa gaagcgatca aagaagcgtc ttcgggcgga gtaggagaat gcggaggaga
      241 aggagaacga gctgatctag tatctctcca caatccaatg ccaactgacc aactggccat
      301 attcggagca atttgaagcc aatttccatc gcctggcgat cgctccattc ttggctatat
      361 gtttttcacc gttacccggg gccattttca aagactcgtc ggcaagataa gattgtgtca
      421 ctcgctgtct ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag
      481 tattttgaag aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac
      541 aaaaacatgt tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt
      601 atgtatatac ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac
      661 tgatgatatt ttaagatgat gccagaccaa aaggcttgaa tttctgcgtc ttttgccgaa
      721 cgcagtgcat gtgcaattgt tgttttttgg aatattcaat tttcggactg tccgctttga
      781 tttcagtttc ttggcttatt caaaaagcaa agtaaagcca aaaaagcgag atggcaatac
      841 caaatgcggc aaaacggtag tggaaggaaa ggggtgcggg gcagcggaag gaagggtggg
      901 gcggggcgtg gcggggtctg tggctgggcg cgacgtcacc gacgttggag ccactccttt
      961 gaccatgtgt gcgtgtgtgt attattcgtg tctcgccact cgccggttgt ttttttcttt
     1021 ttatgctgcg ctctctctag cgccatctcg cttacgcatg ctcaacgcac cgcatgttgc
     1081 cgtttccttt tatgcgtcat tttggctcga aataggcaat tatttaaaca aagattagtc
     1141 aacgaaaacg ctaaaataaa taagtctaca atatggttac ttattgccat gtgtgtgcag
     1201 ccaacgatag caacaaaagc aacaacacag gtggctttcc ctctttcact ttttgtttgc
     1261 aagccgcgtg cgagcaagac ggcacgaccg gcaaacgcaa ttacgctgac aaagagcaga
     1321 cgaagttttg gcgaaaaaca tcaaggcgcc tgatgcgaat gcatttgcaa taacaattgc
     1381 gatatttaat attgtttatg aagctgtttg acttcaaaac acacaaaaaa aaaaataaaa
     1441 caaattattt gaaagagaat taggaatcgg acgcttatcg ttagggtaac aacaagaaat
     1501 gcttactgag tcacagcctc tggaaaactg ccgcaagcca gagagagaga gaaaaagagg
     1561 gagagcagct tagaccgcat gtgcttgtgt gtgaggcgtc tctctcttcg tctctgttgc
     1621 gcaaacgcat agactgcact gaaaaaatcg attacctatt ttttatgaat gaatatttgc
     1681 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca
     1741 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat
     1801 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta
     1861 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgaatgcc ctactagaag
     1921 atgtgtgttg cacaaaatgt ccctggaata accaatttga agtgcagata gcagtaaacg
     1981 taagctaata tgaatattat ttaactgtaa tgttttaata tcgctggaca ttactaataa
     2041 acccactata aacacatgta catatgtatg ttttggcata caatgagtag ttggggaaaa
     2101 aatgtgtaaa agcaccgtga ccatcacagc ataaagataa ccagctgaag tatcgaatat
     2161 gagtaacccc caaattgaat cacatgccgc aactgatagg acccatggaa gtacactctt
     2221 catggcgata tacaagacac acacaagcac gaacacccag ttgcggagga aattctccgt
     2281 aaatgaaaac ccaatcggcg aacaattcat acccatatat ggtaaaagtt ttgaacgcga
     2341 cttgagagcg gagagcattg cggctgataa ggttttagcg ctaagcgggc tttataaaac
     2401 gggctgcggg accagttttc atatcactac cgtttgagtt cttgtgctgt gtggatactc
     2461 ctcccgacac aaagccgctc catcagccag cagtcgtcta atccagagac cccggatcgg
     2521 ggtacccaac atggacgcac aaacacgacg acgtgagcgt cgcgctgaga aacaagctca
     2581 atggaaagct gcaaacggtg gatctcctcc aagcttgatg gcttacccat acgatgttcc
     2641 agattacgct atgcgtgaag cccttttttc ccaagatgcc gcggcctgtc agcatgttaa
     2701 ccaggatact aattgggaag ttcccagttc gccagaacca gccaataagg atgcacccgg
     2761 tccaccaatg tggaagccaa gcattaacaa tggtactgat ctttgggagt ccaatttgag
     2821 aaacggaggt cagccggccg cacagcaagt tccaaagccg tcggcggctc atacaccatc
     2881 ctctaactta ggtggaacag cagctgagga cgacgatggc gccgatagta gtagtgtgtg
     2941 gactggagga gctgttagca acgcgggatc cggagctgca gtgggagtaa accaagccgg
     3001 agttaatgtc ggtccaggcg gtgttgtttc gtctggcgga cctcaggcgg cacaaggtgt
     3061 cgttggcgtc ggacttggat caactggagg taacgggtca agcaatataa ctggatcgtc
     3121 tggagtcgca acaggtagta gcggaaactc cagcaacgct ggtaacgctg cggcagaccc
     3181 tcgtgaaata cgccctttgg gagttggtgg ctccatggat attcgaaatg ttgaacatcg
     3241 cggcggtaac ggttctggag caacttcgag cgatccacga gacattcgca tgatcgatcc
     3301 gcgtgaccct attcgaggag atccccgtgg aatatctggt cgtcttaatg ggacctctga
     3361 aatggccgct catcatccac aaatgtccca taaccagttg caaggtatca acaaaatggt
     3421 tggtcaaagt gtagcaactg ccagcaccag tgtcggaaca tctggctcgg gcatcggtcc
     3481 tggaggtccc ggtcctagta cagtatcagg caatatccca acacaggcgg cgcctgctca
     3541 accggtaagc gttggtgtaa gtggtcccaa agacatgtca aaacagataa gtgcagcgga
     3601 ggaaccatca ccaccgcctc agcgtcgcag tattcctaac tacgatgatg gtacatcgtt
     3661 ggcggctcag caaactcgtg ttcccgctgc aagcggtcac tggaaagaca tgactgattc
     3721 gataggtcgt agtagtcatc tcatgcgtgg ccaaagccaa acgggaggta taggaatagc
     3781 cggcgttgga aatagcaatg ttccagtggg agccaatcca agtaatccta taagcagtgt
     3841 agttggacct caagcccgga ttccatctgt gggcggcgta caacacaaac cagacggcgg
     3901 cgctatgtgg gtgcattccg gcaatgtagg tggcagaaat aatgttgctg ctgttactac
     3961 tgcagcggat gacactcata gcgttaatgt cggcgctccc agcagtggca gtgtatccag
     4021 caacaattgg gttgatgaca agtccaactc aaccttggca caaaactctt ggagcgaccc
     4081 ggcccctgtt ggagttagtt ggggcaataa gcaaagcaaa ccgccaagca atagtgcttc
     4141 atcaggttgg agcactgctg cgggcgtggt ggatggggtt gatctaggat ctgagtggaa
     4201 cacgcacggg gggattattg gaaaatctca gcaacaacaa aaactagcgg gacttaacgt
     4261 gggaatggtg aacgtaatta acgcggagat cattaagcaa agcaagcaat acaggatcct
     4321 tgtcgagaac ggctttaaaa aggaagatgt agagcgggca ttagtgattg ctaatatgaa
     4381 catcgaagag gcagccgata tgctccgtgc caactcatcc ctatcaatgg atggttggcg
     4441 tcgacatgat gagtcccttg gatcttatgc cgaccacaat agttcaacaa gcagcggtgg
     4501 atttgctggt cgttacccgg tcaacagtgg acaaccttca atgtcctttc ctcataataa
     4561 ccttatgaat aacatgggag gtaccgctgt tactggaggt aacaacaata caaacatgac
     4621 agctttacag gtgcaaaagt atttaaatca agggcaacat ggtgtcgctg ttggaccgca
     4681 agccgttggt aattcttcag cagtatctgt cggatttggt cagaacacgt ctaacgcagc
     4741 agtggcagga gcagcctctg caaatatagc agcaaataca aacaaccaac cgtctggtca
     4801 gcaaattcgc atgctaggcc agcaaattca gttggccatt catagtggtt tcatatctag
     4861 tcagatattg actcaaccgc taactcaaac aacccttaac cttttaaacc aacttcttag
     4921 caatattaag catctccagg ctgcacagca atcccttacc cgcgggggaa atgtcaatcc
     4981 aatggcagtg aatgtggcta tatctaaata caagcagcaa atccagaatt tacagaacca
     5041 gataagtgca caacaggctg tgtatgtaaa acagcaaaat atgcaaccaa cttcacaaca
     5101 acaacagccc caacaacagc aacttccttc tgttcatcta agtaactcag gcaacgacta
     5161 tttaagaggt cacgatgcaa taaataattt gcaaagcaac ttttctgagc tcaatattaa
     5221 taagccaagt ggatatcaag gagcgtccaa tcaacaatcc cgattaaatc agtggaagct
     5281 tccagtatta gataaggaga tcaactctga cagtacggaa ttttctcgtg ccccaggtgc
     5341 aacgaaacaa aatttgacgg ccaacacaag caacataaac tctttgggtc ttcaaaacga
     5401 tagtacatgg tcaactggac gcagtattgg tgacggttgg cctgatccct catctgataa
     5461 cgagaataaa gactggtctg ttgctcagcc aacttcagca gcaactgctt acactgatct
     5521 ggtccaagag tttgagccag gcaagccatg gaagggttca cagatcaaaa gcatagaaga
     5581 tgatcccagc attacaccag gaagcgttgc tagatctcca ttgtctatta attcgacgcc
     5641 aaaagatgct gacatatttg ccaataccgg taaaaattca ccgactgatt taccgccact
     5701 aagtttatcg tcgtctacat ggagttttaa tccaaaccaa aattatccga gtcacagttg
     5761 gtctgacaat agtcaacaat gtaccgccac ttcggagctt tggacaagcc cgctaaataa
     5821 atcatcgtct cgaggtcccc cgccaggatt gactgccaat tcaaataaat ctgcaaatag
     5881 taatgcgtca acgccaacaa ctattaccgg aggtgcgaat ggatggttac agcctcgaag
     5941 tggcggtgtt caaaccacaa acactaattg gacaggtggt aacaccactt ggggctccag
     6001 ttggttgctt ttgaaaaatc taacagcaca gattgatggt cctactttgc gtacactgtg
     6061 tatgcagcat gggccccttg tcagctttca cccgtatttg aaccaaggaa ttgccttatg
     6121 taaatatact actcgtgagg aggcgaacaa ggcgcaaatg gcgttaaaca actgtgtcct
     6181 cgccaacacc acaatatttg ctgaatctcc cagcgagaac gaggtgcaaa gcattatgca
     6241 gcacttacca caaactcctt cctctacaag ctctagtgga actagtggtg gcaacgtcgg
     6301 aggcgtcggc acttcagcca ataatgcaaa cagtggttct gcagcttgtc tgtccggaaa
     6361 caatagcggc aacggaaacg gcagcgcgag cggcgccggc agcggcaaca atggcaacag
     6421 tagctgcaac aacagtgccg ccgggggggg cagcagcagc aacaacacga ttaccactgt
     6481 agcaaattcg aatcttgttg gttctagtgg ctctgtctca aattcctctg gcgttactgc
     6541 taactctagt actgtttctg tagttagttg tacagcgagt gggaattcca taaatggggc
     6601 aggtactgca aacagttctg gttcaaagag tagtgcaaac aatttagcta gcggccagtc
     6661 tagcgcttct aacttaacta atagcaccaa ttcaacatgg cgacaaacta gccaaaacca
     6721 agctcttcaa agtcaaagca ggccatcagg cagagaagct gactttgatt atatatctct
     6781 cgtttattcc attgttgatg attaagcggc cgctcgagtc tagagggccc gcggttcgaa
     6841 ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg tcatcatcac
     6901 catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctaa ggcctgagct
     6961 cgctgatcag cctcgatcga ggatccagac atgataagat acattgatga gtttggacaa
     7021 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     7081 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     7141 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     7201 tgtggtatgg ctgattatga tcagtcgacc gatgcccttg agagccttca acccagtcag
     7261 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
     7321 catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt
     7381 tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct
     7441 cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat
     7501 cgccggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg
     7561 gatggccttc cccattatga ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca
     7621 ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag gccagcaaaa
     7681 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7741 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7801 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7861 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7921 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7981 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     8041 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     8101 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     8161 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     8221 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     8281 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     8341 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     8401 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     8461 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     8521 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     8581 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     8641 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     8701 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     8761 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
     8821 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8881 gttgtgcaag aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8941 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     9001 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     9061 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     9121 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     9181 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     9241 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     9301 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     9361 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     9421 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     9481 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     9541 agaattaatt ctatattcta aaaacacaaa tgatacttct aaaaaaaaat
//
