LOCUS       sequence-355951-        7380 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmGW182-MGW-Cterm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7380)
  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 7380)
  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-MGW-Cterm_H
FEATURES             Location/Qualifiers
     source          1..7380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..2510
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2530..2532,2533..2535,2536..3246)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2530..2532,2533..2535,2536..3246)
                     /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(2575..2596)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2836..2855)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3183..3204
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4631..4672
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4682..4699
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4722..4739)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4772..4906
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4796..4815)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4850..4869
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5507..6095)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5587..5606)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6266..7057,7058..7126))
                     /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     6889..6908
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7127..7231)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7299..7317
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 catgaatggc atcaactctg aatcaaatct ttgcagatgc acctacttct catttccact
       61 gtcacatcat ttttccagat ctcgctgcct gttatgtggc ccacaaacca agacacgttt
      121 tatggccatt aaagctggct gatcgtcgcc aaacaccaaa tacataatga atatgtacac
      181 attcgagaaa gaagcgatca aagaagcgtc ttcgggcgga gtaggagaat gcggaggaga
      241 aggagaacga gctgatctag tatctctcca caatccaatg ccaactgacc aactggccat
      301 attcggagca atttgaagcc aatttccatc gcctggcgat cgctccattc ttggctatat
      361 gtttttcacc gttacccggg gccattttca aagactcgtc ggcaagataa gattgtgtca
      421 ctcgctgtct ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag
      481 tattttgaag aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac
      541 aaaaacatgt tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt
      601 atgtatatac ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac
      661 tgatgatatt ttaagatgat gccagaccaa aaggcttgaa tttctgcgtc ttttgccgaa
      721 cgcagtgcat gtgcaattgt tgttttttgg aatattcaat tttcggactg tccgctttga
      781 tttcagtttc ttggcttatt caaaaagcaa agtaaagcca aaaaagcgag atggcaatac
      841 caaatgcggc aaaacggtag tggaaggaaa ggggtgcggg gcagcggaag gaagggtggg
      901 gcggggcgtg gcggggtctg tggctgggcg cgacgtcacc gacgttggag ccactccttt
      961 gaccatgtgt gcgtgtgtgt attattcgtg tctcgccact cgccggttgt ttttttcttt
     1021 ttatgctgcg ctctctctag cgccatctcg cttacgcatg ctcaacgcac cgcatgttgc
     1081 cgtttccttt tatgcgtcat tttggctcga aataggcaat tatttaaaca aagattagtc
     1141 aacgaaaacg ctaaaataaa taagtctaca atatggttac ttattgccat gtgtgtgcag
     1201 ccaacgatag caacaaaagc aacaacacag gtggctttcc ctctttcact ttttgtttgc
     1261 aagccgcgtg cgagcaagac ggcacgaccg gcaaacgcaa ttacgctgac aaagagcaga
     1321 cgaagttttg gcgaaaaaca tcaaggcgcc tgatacgaat gcatttgcaa taacaattgc
     1381 gatatttaat attgtttatg aagctgtttg acttcaaaac acacaaaaaa aaaaataaaa
     1441 caaattattt gaaagagaat taggaatcgg acgcttatcg ttagggtaac aacaagaaat
     1501 gcttactgag tcacagcctc tggaaaactg ccgcaagcca gagagagaga gaaaaagagg
     1561 gagagcagct tagaccgcat gtgcttgtgt gtgaggcgtc tctctcttcg tctctgttgc
     1621 gcaaacgcat agactgcact gaaaaaatcg attacctatt ttttatgaat gaatatttgc
     1681 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca
     1741 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat
     1801 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta
     1861 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgaatgcc ctactagaag
     1921 atgtgtgttg cacaaaatgt ccctggaata accaatttga agtgcagata gcagtaaacg
     1981 taagctaata tgaatattat ttaactgtaa tgttttaata tcgctggaca ttactaataa
     2041 acccactata aacacatgta catatgtatg ttttggcata caatgagtag ttggggaaaa
     2101 aatgtgtaaa agcaccgtga ccatcacagc ataaagataa ccagctgaag tatcgaatat
     2161 gagtaacccc caaattgaat cacatgccgc aactgatagg acccatggaa gtacactctt
     2221 catggcgata tacaagacac acacaagcac gaacacccag ttgcggagga aattctccgt
     2281 aaatgaaaac ccaatcggcg aacaattcat acccatatat ggtaaaagtt ttgaacgcga
     2341 cttgagagcg gagagcattg cggctgataa ggttttagcg ctaagcgggc tttataaaac
     2401 gggctgcggg accagttttc atatcactac cgtttgagtt cttgtgctgt gtggatactc
     2461 ctcccgacac aaagccgctc catcagccag cagtcgtcta atccagagac cccggatcgg
     2521 ggtaccatca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     2581 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     2641 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     2701 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     2761 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     2821 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2881 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2941 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     3001 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     3061 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3121 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     3181 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     3241 tacaagccaa gcttgaattc tgcagatatc cagcacagtg gcggccgcta tcaaggagcg
     3301 tccaatcaac aatcccgatt aaatcagtgg aagcttccag tattagataa ggagatcaac
     3361 tctgacagta cggaattttc tcgtgcccca ggtgcaacga aacaaaattt gacggccaac
     3421 acaagcaaca taaactcttt gggtcttcaa aacgatagta catggtcaac tggacgcagt
     3481 attggtgacg gttggcctga tccctcatct gataacgaga ataaagactg gtctgttgct
     3541 cagccaactt cagcagcaac tgcttacact gatctggtcc aagagtttga gccaggcaag
     3601 ccatggaagg gttcacagat caaaagcata gaagatgatc ccagcattac accaggaagc
     3661 gttgctagat ctccattgtc tattaattcg acgccaaaag atgctgacat atttgccaat
     3721 accggtaaaa attcaccgac tgatttaccg ccactaagtt tatcgtcgtc tacatggagt
     3781 tttaatccaa accaaaatta tccgagtcac agttggtctg acaatagtca acaatgtacc
     3841 gccacttcgg agctttggac aagcccgcta aataaatcat cgtctcgagg tcccccgcca
     3901 ggattgactg ccaattcaaa taaatctgca aatagtaatg cgtcaacgcc aacaactatt
     3961 accggaggtg cgaatggatg gttacagcct cgaagtggcg gtgttcaaac cacaaacact
     4021 aattggacag gtggtaacac cacttggggc ccacaaactc cttcctctac aagctctagt
     4081 ggaactagtg gtggcaacgt cggaggcgtc ggcacttcag ccaataatgc aaacagtggt
     4141 tctgcagctt gtctgtccgg aaacaatagc ggcaacggaa acggcagcgc gagcggcgcc
     4201 ggcagcggca acaatggcaa cagtagctgc aacaacagtg ccgccggggg gggcagcagc
     4261 agcaacaaca cgattaccac tgtagcaaat tcgaatcttg ttggttctag tggctctgtc
     4321 tcaaattcct ctggcgttac tgctaactct agtactgttt ctgtagttag ttgtacagcg
     4381 agtgggaatt ccataaatgg ggcaggtact gcaaacagtt ctggttcaaa gagtagtgca
     4441 aacaatttag ctagcggcca gtctagcgct tctaacttaa ctaatagcac caattcaaca
     4501 tggcgacaaa ctagccaaaa ccaagctctt caaagtcaaa gcaggccatc aggcagagaa
     4561 gctgactttg attatatatc tctcgtttat tccattgttg atgattaatc tagagggccc
     4621 gcggttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg
     4681 tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctaa
     4741 ggcctgagct cgctgatcag cctcgatcga ggatccagac atgataagat acattgatga
     4801 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     4861 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     4921 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     4981 cctctacaaa tgtggtatgg ctgattatga tcagtcgacc gatgcccttg agagccttca
     5041 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
     5101 tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg
     5161 aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct
     5221 tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc
     5281 aggccattat cgccggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga
     5341 cgcgaggctg gatggccttc cccattatga ttcttctcgc ttccggcggc atcgggatgc
     5401 ccgcgttgca ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag
     5461 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5521 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5581 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5641 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5701 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5761 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5821 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5881 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5941 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6001 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6061 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6121 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6181 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6241 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6301 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6361 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6421 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6481 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6541 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac
     6601 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6661 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6721 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6781 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6841 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc
     6901 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6961 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7021 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7081 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7141 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7201 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7261 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     7321 ttcgtcttca agaattaatt ctatattcta aaaacacaaa tgatacttct aaaaaaaaat
//
