LOCUS       sequence-289850-        8723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmGW182-del205-490-dsRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8723)
  AUTHORS   Huntzinger E, Kuzuoglu-Ozturk D, Braun JE, Eulalio A, Wohlbold L, 
            Izaurralde E
  TITLE     The interactions of GW182 proteins with PABP and deadenylases are 
            required for both translational repression and degradation of miRNA 
            targets.
  JOURNAL   Nucleic Acids Res. 2013 Jan;41(2):978-94. doi: 10.1093/nar/gks1078. 
            Epub 2012 Nov 21.
  PUBMED    23172285
REFERENCE   2  (bases 1 to 8723)
  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/gks1078"; journalName: "Nucleic Acids Res"; date: 
            "2013-01"; volume: "41"; issue: "2"; pages: "978-94"
COMMENT     Sequence Label: pAc5.1B-lambdaN-DmGW182-del205-490-dsRNAres_R
FEATURES             Location/Qualifiers
     source          1..8723
                     /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             6008..6049
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6059..6076
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6099..6116)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6149..6283
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6173..6192)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6227..6246
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6905..7493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6985..7004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7664..8455,8456..8524))
                     /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     8287..8306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8525..8629)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8697..8715
                     /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 gcttgatgcg tgaagccctt ttttcccaag
     2701 atggctgggg ctgtcagcat gttaaccagg atactaattg ggaagttccc agttcgccag
     2761 aaccagccaa taaggatgca cccggtccac caatgtggaa gccaagcatt aacaatggta
     2821 ctgatctttg ggagtccaat ttgagaaacg gaggtcagcc ggccgcacag caagttccaa
     2881 agccgtcgtg gggtcataca ccatcctcta acttaggtgg aacatggggt gaggacgacg
     2941 atggcgccga tagtagtagt gtgtggactg gaggagctgt tagcaacgcg ggatccggag
     3001 ctgcagtggg agtaaaccaa gccggagtta atgtcggtcc aggcggtgtt gtttcgtctg
     3061 gcggacctca gtggggacaa ggtgtcgttg gcgtcggact tggatcaact ggcggaaatg
     3121 gttccagtaa cattacaggc tcttcgggcg ttgccacggg aagcagtggc aattcaagta
     3181 atgccggaaa tggatggggc gatccacgcg agattcgtcc actgggcgtc ggaggatcga
     3241 tggacatccg taacgtcgag caccgtggag gaaatggatc aggcgccccg ccaagcaata
     3301 gtgcttcatc aggttggagc actgctgcgg gcgtggtgga tggggttgat ctaggatctg
     3361 agtggaacac gcacgggggg attattggaa aatctcagca acaacaaaaa ctagcgggac
     3421 ttaacgtggg aatggtgaac gtaattaacg cggagatcat taagcaaagc aagcaataca
     3481 ggatccttgt cgagaacggc tttaaaaagg aagatgtaga gcgggcatta gtgattgcta
     3541 atatgaacat cgaagaggca gccgatatgc tccgtgccaa ctcatcccta tcaatggatg
     3601 gttggcgtcg acatgatgag tcccttggat cttatgccga ccacaatagt tcaacaagca
     3661 gcggtggatt tgctggtcgt tacccggtca acagtggaca accttcaatg tcctttcctc
     3721 ataataacct tatgaataac atgggaggta ccgctgttac tggaggtaac aacaatacaa
     3781 acatgacagc tttacaggtg caaaagtatt taaatcaagg gcaacatggt gtcgctgttg
     3841 gaccgcaagc cgttggtaat tcttcagcag tatctgtcgg atttggtcag aacacgtcta
     3901 acgcagcagt ggcaggagca gcctctgcaa atatagcagc aaatacaaac aaccaaccgt
     3961 ctggtcagca aattcgcatg ctaggccagc aaattcagtt ggccattcat agtggtttca
     4021 tatctagtca gatattgact caaccgctaa ctcaaacaac ccttaacctt ttaaaccaac
     4081 ttcttagcaa tattaagcat ctccaggctg cacagcaatc ccttacccgc gggggaaatg
     4141 tcaatccaat ggcagtgaat gtggctatat ctaaatacaa gcagcaaatc cagaatttac
     4201 agaaccagat aagtgcacaa caggctgtgt atgtaaaaca gcaaaatatg caaccaactt
     4261 cacaacaaca acagccccaa caacagcaac ttccttctgt tcatctaagt aactcaggca
     4321 acgactattt aagaggtcac gatgcaataa ataatttgca aagcaacttt tctgagctca
     4381 atattaataa gccaagtgga tatcaaggag cgtccaatca acaatcccga ttaaatcagt
     4441 ggaagcttcc agtattagat aaggagatca actctgacag tacggaattt tctcgtgccc
     4501 caggtgcaac gaaacaaaat ttgacggcca acacaagcaa cataaactct ttgggtcttc
     4561 aaaacgatag tacatggtca actggacgca gtattggtga cggttggcct gatccctcat
     4621 ctgataacga gaataaagac tggtctgttg ctcagccaac ttcagcagca actgcttaca
     4681 ctgatctggt ccaagagttt gagccaggca agccatggaa gggttcacag atcaaaagca
     4741 tagaagatga tcccagcatt acaccaggaa gcgttgctag atctccattg tctattaatt
     4801 cgacgccaaa agatgctgac atatttgcca ataccggtaa aaattcaccg actgatttac
     4861 cgccactaag tttatcgtcg tctacatgga gttttaatcc aaaccaaaat tatccgagtc
     4921 acagttggtc tgacaatagt caacaatgta ccgccacttc ggagctttgg acaagcccgc
     4981 taaataaatc atcgtctcga ggtcccccgc caggattgac tgccaattca aataaatctg
     5041 caaatagtaa tgcgtcaacg ccaacaacta ttaccggagg tgcgaatgga tggttacagc
     5101 ctcgaagtgg cggtgttcaa accacaaaca ctaattggac aggtggtaac accacttggg
     5161 gctccagttg gttgcttttg aaaaatctaa cagcacagat tgatggtcct actttgcgta
     5221 cactgtgtat gcagcatggg ccccttgtca gctttcaccc gtatttgaac caaggaattg
     5281 ccttatgtaa atatactact cgtgaggagg cgaacaaggc gcaaatggcg ttaaacaact
     5341 gtgtcctcgc caacaccaca atatttgctg aatctcccag cgagaacgag gtgcaaagca
     5401 ttatgcagca cttaccacaa actccttcct ctacaagctc tagtggaact agtggtggca
     5461 acgtcggagg cgtcggcact tcagccaata atgcaaacag tggttctgca gcttgtctgt
     5521 ccggaaacaa tagcggcaac ggaaacggca gcgcgagcgg cgccggcagc ggcaacaatg
     5581 gcaacagtag ctgcaacaac agtgccgccg gggggggcag cagcagcaac aacacgatta
     5641 ccactgtagc aaattcgaat cttgttggtt ctagtggctc tgtctcaaat tcctctggcg
     5701 ttactgctaa ctctagtact gtttctgtag ttagttgtac agcgagtggg aattccataa
     5761 atggggcagg tactgcaaac agttctggtt caaagagtag tgcaaacaat ttagctagcg
     5821 gccagtctag cgcttctaac ttaactaata gcaccaattc aacatggcga caaactagcc
     5881 aaaaccaagc tcttcaaagt caaagcaggc catcaggcag agaagctgac tttgattata
     5941 tatctctcgt ttattccatt gttgatgatt aagcggccgc tcgagtctag agggcccgcg
     6001 gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca
     6061 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctaaggc
     6121 ctgagctcgc tgatcagcct cgatcgagga tccagacatg ataagataca ttgatgagtt
     6181 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     6241 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     6301 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     6361 ctacaaatgt ggtatggctg attatgatca gtcgaccgat gcccttgaga gccttcaacc
     6421 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
     6481 tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg
     6541 accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc
     6601 acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg
     6661 ccattatcgc cggcatggcg gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc
     6721 gaggctggat ggccttcccc attatgattc ttctcgcttc cggcggcatc gggatgcccg
     6781 cgttgcaggc catgctgtcc aggcaggtag atgacgacca tcagggacag cttcaaggat
     6841 cgctcgcggc tcttaccagc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6901 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6961 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     7021 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     7081 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     7141 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     7201 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7261 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7321 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     7381 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7441 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7501 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7561 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7621 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7681 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7741 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7801 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7861 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7921 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgctg
     7981 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     8041 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     8101 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     8161 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     8221 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8281 cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8341 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8401 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8461 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8521 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8581 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8641 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     8701 ggcgtatcac gaggcccttt cgt
//
