LOCUS       sequence-289849-        9381 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 9381)
  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 9381)
  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-EGFP-DmGW182-del205-490-dsRNAres_R
FEATURES             Location/Qualifiers
     source          1..9381
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2568
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2588..2590,2591..2593,2594..3304)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2588..2590,2591..2593,2594..3304)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2633..2654)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2894..2913)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3241..3262
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6666..6707
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6717..6734
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6757..6774)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6807..6941
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6831..6850)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6885..6904
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7563..8151)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7643..7662)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8322..9113,9114..9182))
                     /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     8945..8964
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9183..9287)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9355..9373
                     /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 taccaacatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     2641 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     2701 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     2761 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     2821 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     2881 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     2941 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     3001 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     3061 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     3121 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     3181 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     3241 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     3301 caagccaagc ttgaattctg cagatatcca gcacagtggc ggccgcatgc gtgaagccct
     3361 tttttcccaa gatggctggg gctgtcagca tgttaaccag gatactaatt gggaagttcc
     3421 cagttcgcca gaaccagcca ataaggatgc acccggtcca ccaatgtgga agccaagcat
     3481 taacaatggt actgatcttt gggagtccaa tttgagaaac ggaggtcagc cggccgcaca
     3541 gcaagttcca aagccgtcgt ggggtcatac accatcctct aacttaggtg gaacatgggg
     3601 tgaggacgac gatggcgccg atagtagtag tgtgtggact ggaggagctg ttagcaacgc
     3661 gggatccgga gctgcagtgg gagtaaacca agccggagtt aatgtcggtc caggcggtgt
     3721 tgtttcgtct ggcggacctc agtggggaca aggtgtcgtt ggcgtcggac ttggatcaac
     3781 tggcggaaat ggttccagta acattacagg ctcttcgggc gttgccacgg gaagcagtgg
     3841 caattcaagt aatgccggaa atggatgggg cgatccacgc gagattcgtc cactgggcgt
     3901 cggaggatcg atggacatcc gtaacgtcga gcaccgtgga ggaaatggat caggcgcccc
     3961 gccaagcaat agtgcttcat caggttggag cactgctgcg ggcgtggtgg atggggttga
     4021 tctaggatct gagtggaaca cgcacggggg gattattgga aaatctcagc aacaacaaaa
     4081 actagcggga cttaacgtgg gaatggtgaa cgtaattaac gcggagatca ttaagcaaag
     4141 caagcaatac aggatccttg tcgagaacgg ctttaaaaag gaagatgtag agcgggcatt
     4201 agtgattgct aatatgaaca tcgaagaggc agccgatatg ctccgtgcca actcatccct
     4261 atcaatggat ggttggcgtc gacatgatga gtcccttgga tcttatgccg accacaatag
     4321 ttcaacaagc agcggtggat ttgctggtcg ttacccggtc aacagtggac aaccttcaat
     4381 gtcctttcct cataataacc ttatgaataa catgggaggt accgctgtta ctggaggtaa
     4441 caacaataca aacatgacag ctttacaggt gcaaaagtat ttaaatcaag ggcaacatgg
     4501 tgtcgctgtt ggaccgcaag ccgttggtaa ttcttcagca gtatctgtcg gatttggtca
     4561 gaacacgtct aacgcagcag tggcaggagc agcctctgca aatatagcag caaatacaaa
     4621 caaccaaccg tctggtcagc aaattcgcat gctaggccag caaattcagt tggccattca
     4681 tagtggtttc atatctagtc agatattgac tcaaccgcta actcaaacaa cccttaacct
     4741 tttaaaccaa cttcttagca atattaagca tctccaggct gcacagcaat cccttacccg
     4801 cgggggaaat gtcaatccaa tggcagtgaa tgtggctata tctaaataca agcagcaaat
     4861 ccagaattta cagaaccaga taagtgcaca acaggctgtg tatgtaaaac agcaaaatat
     4921 gcaaccaact tcacaacaac aacagcccca acaacagcaa cttccttctg ttcatctaag
     4981 taactcaggc aacgactatt taagaggtca cgatgcaata aataatttgc aaagcaactt
     5041 ttctgagctc aatattaata agccaagtgg atatcaagga gcgtccaatc aacaatcccg
     5101 attaaatcag tggaagcttc cagtattaga taaggagatc aactctgaca gtacggaatt
     5161 ttctcgtgcc ccaggtgcaa cgaaacaaaa tttgacggcc aacacaagca acataaactc
     5221 tttgggtctt caaaacgata gtacatggtc aactggacgc agtattggtg acggttggcc
     5281 tgatccctca tctgataacg agaataaaga ctggtctgtt gctcagccaa cttcagcagc
     5341 aactgcttac actgatctgg tccaagagtt tgagccaggc aagccatgga agggttcaca
     5401 gatcaaaagc atagaagatg atcccagcat tacaccagga agcgttgcta gatctccatt
     5461 gtctattaat tcgacgccaa aagatgctga catatttgcc aataccggta aaaattcacc
     5521 gactgattta ccgccactaa gtttatcgtc gtctacatgg agttttaatc caaaccaaaa
     5581 ttatccgagt cacagttggt ctgacaatag tcaacaatgt accgccactt cggagctttg
     5641 gacaagcccg ctaaataaat catcgtctcg aggtcccccg ccaggattga ctgccaattc
     5701 aaataaatct gcaaatagta atgcgtcaac gccaacaact attaccggag gtgcgaatgg
     5761 atggttacag cctcgaagtg gcggtgttca aaccacaaac actaattgga caggtggtaa
     5821 caccacttgg ggctccagtt ggttgctttt gaaaaatcta acagcacaga ttgatggtcc
     5881 tactttgcgt acactgtgta tgcagcatgg gccccttgtc agctttcacc cgtatttgaa
     5941 ccaaggaatt gccttatgta aatatactac tcgtgaggag gcgaacaagg cgcaaatggc
     6001 gttaaacaac tgtgtcctcg ccaacaccac aatatttgct gaatctccca gcgagaacga
     6061 ggtgcaaagc attatgcagc acttaccaca aactccttcc tctacaagct ctagtggaac
     6121 tagtggtggc aacgtcggag gcgtcggcac ttcagccaat aatgcaaaca gtggttctgc
     6181 agcttgtctg tccggaaaca atagcggcaa cggaaacggc agcgcgagcg gcgccggcag
     6241 cggcaacaat ggcaacagta gctgcaacaa cagtgccgcc ggggggggca gcagcagcaa
     6301 caacacgatt accactgtag caaattcgaa tcttgttggt tctagtggct ctgtctcaaa
     6361 ttcctctggc gttactgcta actctagtac tgtttctgta gttagttgta cagcgagtgg
     6421 gaattccata aatggggcag gtactgcaaa cagttctggt tcaaagagta gtgcaaacaa
     6481 tttagctagc ggccagtcta gcgcttctaa cttaactaat agcaccaatt caacatggcg
     6541 acaaactagc caaaaccaag ctcttcaaag tcaaagcagg ccatcaggca gagaagctga
     6601 ctttgattat atatctctcg tttattccat tgttgatgat taatctagag ggcccgcggt
     6661 tcgaaggtaa gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggtcatc
     6721 atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct tctaaggcct
     6781 gagctcgctg atcagcctcg atcgaggatc cagacatgat aagatacatt gatgagtttg
     6841 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     6901 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     6961 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     7021 acaaatgtgg tatggctgat tatgatcagt cgaccgatgc ccttgagagc cttcaaccca
     7081 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
     7141 tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac
     7201 cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac
     7261 gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc
     7321 attatcgccg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga
     7381 ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg
     7441 ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg
     7501 ctcgcggctc ttaccagcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     7561 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     7621 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     7681 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     7741 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7801 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7861 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7921 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7981 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     8041 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     8101 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     8161 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     8221 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     8281 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     8341 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     8401 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     8461 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     8521 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     8581 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca
     8641 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     8701 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     8761 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     8821 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     8881 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
     8941 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     9001 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     9061 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     9121 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     9181 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     9241 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     9301 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     9361 cgtatcacga ggccctttcg t
//
