LOCUS       sequence-289788-        7809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmNot2-dsRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7809)
  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 7809)
  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-DmNot2-dsRNAres_Q
FEATURES             Location/Qualifiers
     source          1..7809
                     /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             5094..5135
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5145..5162
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5185..5202)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5235..5369
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5259..5278)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5313..5332
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5991..6579)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6071..6090)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6750..7541,7542..7610))
                     /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     7373..7392
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7611..7715)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7783..7801
                     /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 ttgatggcga atttaaattt tcaacaaccc ccgagaagta tagcgaacgc
     3361 ggcgttagct ggccggacaa caggcggatt cggtggtagc tccctggctg gccatgtgac
     3421 gcccacgtcg ggcatgtttc aaaccgactt cgccaactcc tatccaggaa cggcgaacta
     3481 cggacaggcg ccgcagcagc agcaacaaca gcagcagcag ccccaactgt cgccgaaccg
     3541 gaatgcccag ctgtcggtcg gcggacccgc catctcgagc ggcaaccgca acgccaacct
     3601 cttcggccag cgtcagtttg tggagcggcg agccatgcaa ggattgggca gtggccccat
     3661 gtcgaacatg ggcaacttca tgcagacggg tcgcggcggc tacggaacgg gcggaggcgg
     3721 tggcggtcct ttgaataact tccacgtgtt cggcggcggc ggtggtagcg acacctccac
     3781 gcctgccctg ctcgatccca cggagttccc ctccttgacc aacgctcgtg gacaaaatga
     3841 tcaaacgttg ccgcagtcca accccttgca accccccgga tccaagccgt acggaaactt
     3901 tttcacgtca ttcggaatgg tcaaacagcc caccagtgaa cagagcgaat ttaccatgtc
     3961 aaatgaagat tttccagccc ttcccggaac gcagaattcc gatggaacga ccaatgccgt
     4021 cggttccgtg gctggcacgg gcggaagcgg cggagctagc acggaaaatc atctggatgg
     4081 aaccgagaaa cccatgaact ccatcgtcgt gtccggcagt gcctccggat cctccggaag
     4141 caatgtggga gtcgtcggag gaaacggact gggagctgtc ggatcgggaa ttggtggact
     4201 ggcggtcgga ggaggaggcg gcgcgggatc cagtggcgga ggcggcgtgg gaggaaacgc
     4261 ggcttccgga gtcgtgggag gtagccacgt tggcttggtc ggatcgaatt ccggaatcgg
     4321 aggagtgaat tccgttccga attccaacgc tatgatggga gttggaggag gattgggttc
     4381 gggctccgga tcctcaggtt ccggcgccgg aggagaacac ttgaatgaca acagctccaa
     4441 tgacaagctc gttaaatccg gagtccagac atccccggac ggtaaagtga cgaatattcc
     4501 ggccacgatg gtcaataatc aatttggaat ggtcggattg ttgaccttta tccgcgcggc
     4561 ggagaccgac ccgaacttgg tcaccttgag cttgggtacg gatctcacgg gtttaggcct
     4621 gaacctgaac tcgcaggaga gcttgcatac aaccttcgct ggccccttcg tagcacagcc
     4681 ctgcagagca caagacgttg agttcaacgt gccacccgaa tatctgatta actttgcgat
     4741 aagagacaaa ctaacagcgc cggtactgaa aaagctgcaa gaagacctgc tctttttcct
     4801 tttttacacg aacatcggcg atatgatgca attgatggca gctgctgaat tgcatagtcg
     4861 cgagtggcga taccacgtgg aggagaaaat atggataact cgaattcctg gcattgatca
     4921 atatgaaaaa aatggtacaa aagagcgcgg caccttctac tactttgatg cgcaaagttg
     4981 gaaacgtttg tccaaggttt tccaaataga tcccgagaaa ttagataaat gtcccaatat
     5041 aagtgcgttt atgaatggac agtctgtata atctagaggg cccgcggttc gaaggtaagc
     5101 ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc
     5161 attgagttta aacccgctga tcagcctcga ctgtgccttc taaggcctga gctcgctgat
     5221 cagcctcgat cgaggatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     5281 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     5341 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     5401 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     5461 tggctgatta tgatcagtcg accgatgccc ttgagagcct tcaacccagt cagctccttc
     5521 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     5581 ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg
     5641 agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa
     5701 gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc
     5761 atggcggccg acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc
     5821 ttccccatta tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg
     5881 ctgtccaggc aggtagatga cgaccatcag ggacagcttc aaggatcgct cgcggctctt
     5941 accagccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6001 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6061 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6121 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6181 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6241 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6301 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6361 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6421 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6481 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6541 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6601 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6661 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6721 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6781 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6841 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6901 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6961 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7021 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg
     7081 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7141 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7201 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7261 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7321 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc
     7381 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7441 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7501 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7561 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7621 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7681 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7741 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     7801 ccctttcgt
//
