LOCUS       sequence-288878-        9684 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmGW182-delCterm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9684)
  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 9684)
  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-EGFP-DmGW182-delCterm_G
FEATURES             Location/Qualifiers
     source          1..9684
                     /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             6969..7010
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             7020..7037
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7060..7077)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7110..7244
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7134..7153)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7188..7207
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7866..8454)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7946..7965)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8625..9416,9417..9485))
                     /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     9248..9267
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9486..9590)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9658..9676
                     /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 tggaggtaac gggtcaagca atataactgg atcgtctgga gtcgcaacag gtagtagcgg
     3841 aaactccagc aacgctggta acggttgggg agaccctcgt gaaatacgcc ctttgggagt
     3901 tggtggctcc atggatattc gaaatgttga acatcgcggc ggtaacggtt ctggagcaac
     3961 ttcgagcgat ccacgagaca ttcgcatgat cgatccgcgt gaccctattc gaggagatcc
     4021 ccgtggaata tctggtcgtc ttaatgggac ctctgaaatg tggggtcatc atccacaaat
     4081 gtcccataac cagttgcaag gtatcaacaa aatggttggt caaagtgtag caactgccag
     4141 caccagtgtc ggaacatctg gctcgggcat cggtcctgga ggtcccggtc ctagtacagt
     4201 atcaggcaat atcccaacac agtgggggcc tgctcaaccg gtaagcgttg gtgtaagtgg
     4261 tcccaaagac atgtcaaaac agataagtgg atgggaggaa ccatcaccac cgcctcagcg
     4321 tcgcagtatt cctaactacg atgatggtac atcgttgtgg ggtcagcaaa ctcgtgttcc
     4381 cgctgcaagc ggtcactgga aagacatgac tgattcgata ggtcgtagta gtcatctcat
     4441 gcgtggccaa agccaaacgg gaggtatagg aatagccggc gttggaaata gcaatgttcc
     4501 agtgggagcc aatccaagta atcctataag cagtgtagtt ggacctcaag cccggattcc
     4561 atctgtgggc ggcgtacaac acaaaccaga cggcggcgct atgtgggtgc attccggcaa
     4621 tgtaggtggc agaaataatg ttgctgctgt tactacttgg ggagatgaca ctcatagcgt
     4681 taatgtcggc gctcccagca gtggcagtgt atccagcaac aattgggttg atgacaagtc
     4741 caactcaacc ttggcacaaa actcttggag cgacccggcc cctgttggag ttagttgggg
     4801 caataagcaa agcaaaccgc caagcaatag tgcttcatca ggttggagca ctgctgcggg
     4861 cgtggtggat ggggttgatc taggatctga gtggaacacg cacgggggga ttattggaaa
     4921 atctcagcaa caacaaaaac tagcgggact taacgtggga atggtgaacg taattaacgc
     4981 ggagatcatt aagcaaagca agcaatacag gatccttgtc gagaacggct ttaaaaagga
     5041 agatgtagag cgggcattag tgattgctaa tatgaacatc gaagaggcag ccgatatgct
     5101 ccgtgccaac tcatccctat caatggatgg ttggcgtcga catgatgagt cccttggatc
     5161 ttatgccgac cacaatagtt caacaagcag cggtggattt gctggtcgtt acccggtcaa
     5221 cagtggacaa ccttcaatgt cctttcctca taataacctt atgaataaca tgggaggtac
     5281 cgctgttact ggaggtaaca acaatacaaa catgacagct ttacaggtgc aaaagtattt
     5341 aaatcaaggg caacatggtg tcgctgttgg accgcaagcc gttggtaatt cttcagcagt
     5401 atctgtcgga tttggtcaga acacgtctaa cgcagcagtg gcaggagcag cctctgtaaa
     5461 tatagcagca aatacaaaca accaaccgtc tggtcagcaa attcgcatgc taggccagca
     5521 aattcagttg gccattcata gtggtttcat atctagtcag atattgactc aaccgctaac
     5581 tcaaacaacc cttaaccttt taaaccaact tcttagcaat attaagcatc tccaggctgc
     5641 gcagcaatcc cttacccgcg ggggaaatgt caatccaatg gcagtgaatg tggctatatc
     5701 taaatacaag cagcaaatcc agaatttaca gaaccagata aatgcacaac aggctgtgta
     5761 tgtaaaacag caaaatatgc aaccaacttc acaacaacaa cagccccaac aacagcaact
     5821 tccttctgtt catctaagta actcaggcaa cgactattta agaggtcacg atgcaataaa
     5881 taatttgcaa agcaactttt ctgagctcaa tattaataag ccaagtggat atcaaggagc
     5941 gtccaatcaa caatcccgat taaatcagtg gaagcttcca gtattagata aggagatcaa
     6001 ctctgacagt acggaatttt ctcgtgcccc aggtgcaacg aaacaaaatt tgacggccaa
     6061 cacaagcaac ataaactctt tgggtcttca aaacgatagt acatggtcaa ctggacgcag
     6121 tattggtgac ggttggcctg atccctcatc tgataacgag aataaagact ggtctgttgc
     6181 tcagccaact tcagcagcaa ctgcttacac tgatctggtc caagagtttg agccaggcaa
     6241 gccatggaag ggttcacaga tcaaaagcat agaagatgat cccagcatta caccaggaag
     6301 cgttgctaga tctccattgt ctattaattc gacgccaaaa gatgctgaca tatttgccaa
     6361 taccggtaaa aattcaccga ctgatttacc gccactaagt ttatcgtcgt ctacatggag
     6421 ttttaatcca aaccaaaatt atccgagtca cagttggtct gacaatagtc aacaatgtac
     6481 cgccacttcg gagctttgga caagcccgct aaataaatca tcgtctcgag gtcccccgcc
     6541 aggattgact gccaattcaa ataaatctgc aaatagtaat gcgtcaacgc caacaactat
     6601 taccggaggt gcgaatggat ggttacagcc tcgaagtggc ggtgttcaaa ccacaaacac
     6661 taattggaca ggtggtaaca ccacttgggg ctccagttgg ttgcttttga aaaatctaac
     6721 agcacagatt gatggtccta ctttgcgtac actgtgtatg cagcatgggc cccttgtcag
     6781 ctttcacccg tatttgaacc aaggaattgc cttatgtaaa tatactactc gtgaggaggc
     6841 gaacaaggcg caaatggcgt taaacaactg tgtcctcgcc aacaccacaa tatttgctga
     6901 atctcccagc gagaacgagg tgcaaagcat tatgcagcac ttataatcta gagggcccgc
     6961 ggttcgaagg taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtc
     7021 atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg ccttctaagg
     7081 cctgagctcg ctgatcagcc tcgatcgagg atccagacat gataagatac attgatgagt
     7141 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     7201 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     7261 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     7321 tctacaaatg tggtatggct gattatgatc agtcgaccga tgcccttgag agccttcaac
     7381 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
     7441 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag
     7501 gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg
     7561 cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag
     7621 gccattatcg ccggcatggc ggccgacgcg ctgggctacg tcttgctggc gttcgcgacg
     7681 cgaggctgga tggccttccc cattatgatt cttctcgctt ccggcggcat cgggatgccc
     7741 gcgttgcagg ccatgctgtc caggcaggta gatgacgacc atcagggaca gcttcaagga
     7801 tcgctcgcgg ctcttaccag ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     7861 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     7921 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     7981 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     8041 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     8101 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     8161 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     8221 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     8281 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     8341 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     8401 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     8461 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     8521 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     8581 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     8641 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     8701 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     8761 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     8821 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     8881 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     8941 gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     9001 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     9061 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     9121 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     9181 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     9241 acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     9301 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     9361 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     9421 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     9481 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     9541 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     9601 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     9661 aggcgtatca cgaggccctt tcgt
//
