LOCUS       sequence-288926-        7179 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmGW182-DUF-M2-Cterm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7179)
  AUTHORS   Zekri L, Huntzinger E, Heimstadt S, Izaurralde E
  TITLE     The silencing domain of GW182 interacts with PABPC1 to promote 
            translational repression and degradation of microRNA targets and is 
            required for target release.
  JOURNAL   Mol Cell Biol. 2009 Dec;29(23):6220-31. doi: 10.1128/MCB.01081-09. 
            Epub 2009 Sep 21.
  PUBMED    19797087
REFERENCE   2  (bases 1 to 7179)
  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: 
            "10.1128/MCB.01081-09"; journalName: "Mol Cell Biol"; date: 
            "2009-12"; volume: "29"; issue: "23"; pages: "6220-31"
COMMENT     Sequence Label: pAc5.1B-EGFP-DmGW182-DUF-M2-Cterm_H
FEATURES             Location/Qualifiers
     source          1..7179
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1991..1993,1994..1996,1997..2707)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1991..1993,1994..1996,1997..2707)
                     /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(2036..2057)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2297..2316)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2644..2665
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3867..3908
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3918..3935
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3958..3975)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4008..4142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4032..4051)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4086..4105
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4764..5352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4844..4863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5523..6314,6315..6383))
                     /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     6146..6165
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6384..6488)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6556..6574
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(6638..7179,1..1971)
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
ORIGIN
        1 tgttatctat agataatttt gttgcaaaat atgttgacta tgacaaagat tgtatgtata
       61 tacctttaat gtattctcat tttcttatgt atttataatg gcaatgatga tactgatgat
      121 attttaagat gatgccagac cacaggctga tttctgcgtc ttttgccgaa cgcagtgcat
      181 gtgcggttgt tgttttttgg aatagtttca attttcggac tgtccgcttt gatttcagtt
      241 tcttggctta ttcaaaaagc aaagtaaagc caaaaaagcg agatggcaat accaaatgcg
      301 gcaaaacggt agtggaagga aaggggtgcg gggcagcgga aggaagggtg gggcggggcg
      361 tggcggggtc tgtggctggg cgcgacgtca ccgacgttgg agccactcct ttgaccatgt
      421 gtgcgtgtgt gtattattcg tgtctcgcca ctcgccggtt gtttttttct ttttatctcg
      481 ctctctctag cgccatctcg tacgcatgct caacgcaccg catgttgccg tgtcctttat
      541 gcgtcatttt ggctcgaaat aggcaattat ttaaacaaag attagtcaac gaaaacgcta
      601 aaataaataa gtctacaata tggttactta ttgccatgtg tgtgcagcca acgatagcaa
      661 caaaagcaac aacacagtgg ctttccctct ttcacttttt gtttgcaagc gcgtgcgagc
      721 aagacggcac gaccggcaaa cgcaattacg ctgacaaaga gcagacgaag ttttggccga
      781 aaaacatcaa ggcgcctgat acgaatgcat ttgcaataac aattgcgata tttaatattg
      841 tttatgaagc tgtttgactt caaaacacac aaaaaaaaaa ataaaacaaa ttatttgaaa
      901 gagaattagg aatcggacag cttatcgtta cgggctaaca gcacaccgag acgaaatagc
      961 ttacctgacg tcacagcctc tggaagaact gccgccaagc agagagagag agaaaaagag
     1021 ggagagcagc ttagaccgca tgtgcttgtg tgtgaggcgt ctctctcttc gtctcctgtt
     1081 tgcgcaaacg catagactgc actgagaaaa tcgattacct attttttatg aatgaatatt
     1141 tgcactatta ctattcaaaa ctattaagat agcaatcaca ttcaatagcc aaatactata
     1201 ccacctgagc gatgcaacga aatgatcaat ttgagcaaaa atgctgcata tttaggacgg
     1261 catcattata gaaatgcttc ttgctgtgta cttttctctc gtctggcagc tgtttcgccg
     1321 ttattgttaa aaccggctta agttaggtgt gttttctacg actagtgatg cccctactag
     1381 aagatgtgtg ttgcacaaat gtccctgaat aaccaatttg aagtgcagat agcagtaaac
     1441 gtaagctaat atgaatatta tttaactgta atgttttaat atcgctggac attactaata
     1501 aacccactat aaacacatgt acatatgtat gttttggcat acaatgagta gttggggaaa
     1561 aaatgtgtaa aagcaccgtg accatcacag cataaagata accagctgaa gtatcgaata
     1621 tgagtaaccc ccaaattgaa tcacatgccg caactgatag gacccatgga agtacactct
     1681 tcatggcgat atacaagaca cacacaagca cgaacaccca gttgcggagg aaattctccg
     1741 taaatgaaaa cccaatcggc gaacaattca tacccatata tggtaaaagt tttgaacgcg
     1801 acttgagagc ggagagcatt gcggctgata aggttttagc gctaagcggg ctttataaaa
     1861 cgggctgcgg gaccagtttt catatcacta ccgtttgagt tcttgtgctg tgtggatact
     1921 cctcccgaca caaagccgct ccatcagcca gcagtcgtct aatccagaga ccccggatcg
     1981 gggtaccaac atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     2041 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     2101 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     2161 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     2221 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2281 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2341 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2401 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     2461 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     2521 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2581 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     2641 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2701 gtacaagcca agcttgaatt ctgcagatat ccagcacagt ggcggccgca acgagaataa
     2761 agactggtct gttgctcagc caacttcagc agcaactgct tacactgatc tggtccaaga
     2821 gtttgagcca ggcaagccat ggaagggttc acagatcaaa agcatagaag atgatcccag
     2881 cattacacca ggaagcgttg ctagatctcc attgtctatt aattcgacgc caaaagatgc
     2941 tgacatattt gccaataccg gtaaaaattc accgactgat ttaccgccac taagtttatc
     3001 gtcgtctaca tggagtttta atccaaacca aaattatccg agtcacagtt ggtctgacaa
     3061 tagtcaacaa tgtaccgcca cttcggagct ttggacaagc ccgctaaata aatcatcgtc
     3121 tcgaggtccc ccgccaggat tgactgccaa ttcaaataaa tctgcaaata gtaatgcgtc
     3181 aacgccaaca actattaccg gaggtgcgaa tggatggtta cagcctcgaa gtggcggtgt
     3241 tcaaaccaca aacactaatt ggacaggtgg taacaccact tggggcccac aaactccttc
     3301 ctctacaagc tctagtggaa ctagtggtgg caacgtcgga ggcgtcggca cttcagccaa
     3361 taatgcaaac agtggttctg cagcttgtct gtccggaaac aatagcggca acggaaacgg
     3421 cagcgcgagc ggcgccggca gcggcaacaa tggcaacagt agctgcaaca acagtgccgc
     3481 cggggggggc agcagcagca acaacacgat taccactgta gcaaattcga atcttgttgg
     3541 ttctagtggc tctgtctcaa attcctctgg cgttactgct aactctagta ctgtttctgt
     3601 agttagttgt acagcgagtg ggaattccat aaatggggca ggtactgcaa acagttctgg
     3661 ttcaaagagt agtgcaaaca atttagctag cggccagtct agcgcttcta acttaactaa
     3721 tagcaccaat tcaacatggc gacaaactag ccaaaaccaa gctcttcaaa gtcaaagcag
     3781 gccatcaggc agagaagctg actttgatta tatatctctc gtttattcca ttgttgatga
     3841 ttaatctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg
     3901 attctacgcg taccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct
     3961 cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga
     4021 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     4081 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     4141 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     4201 tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgaccgatg
     4261 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
     4321 gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc
     4381 tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt
     4441 gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa
     4501 cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc
     4561 ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct tctcgcttcc
     4621 ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat
     4681 cagggacagc ttcaaggatc gctcgcggct cttaccagcc agcaaaaggc caggaaccgt
     4741 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4801 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4861 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4921 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     4981 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5041 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5101 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5161 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     5221 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5281 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5341 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5401 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5461 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5521 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5581 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5641 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5701 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5761 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5821 aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5881 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5941 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6001 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6061 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6121 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     6181 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6241 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6301 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6361 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6421 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6481 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     6541 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaagg aattaattct
     6601 atattctaaa aacacaaatg atacttctaa aaaaaatcat gaatggcatc aactctgaat
     6661 caaatctttg cagatgcacc tacttctcat ttccactgtc acatcatttt tccagatctc
     6721 gctgcctgtt atgtggccca caaaccaaga cacgttttat ggccattaaa gctggctgat
     6781 cgtcgccaaa caccaaatac atatcaatat gtacattcga gaaagaagcg atcaaagaag
     6841 cgtcttcggg cgagtaggag aatgcggagg agaaggagaa cgagctgatc tagtatctct
     6901 ccacaatcca atgccaactg accaactggc catattcgga gcaatttgaa gccaatttcc
     6961 atcgcctggc gatcgctcca ttcttggcta tatgtttttc accgttcccg gggccatttt
     7021 caaagactcg tcggtaagat aagattgtgt cactcgctgt ctctcttcat ttgtcgaaga
     7081 atgctgagga atttcgcgat gacgtcggcg agtattttga agaatgagaa taatttgtat
     7141 ttatacgaaa atcagttagt ggaattttct acaaaaaca
//
