LOCUS       sequence-288930-        7656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmGW182-MGW-RRM-Cterm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7656)
  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 7656)
  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-RRM-Cterm_H
FEATURES             Location/Qualifiers
     source          1..7656
                     /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             4344..4385
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4395..4412
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4435..4452)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4485..4619
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4509..4528)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4563..4582
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5241..5829)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5321..5340)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6000..6791,6792..6860))
                     /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     6623..6642
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6861..6965)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7033..7051
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(7115..7656,1..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 ggcggccgct atcaaggagc
     2761 gtccaatcaa caatcccgat taaatcagtg gaagcttcca gtattagata aggagatcaa
     2821 ctctgacagt acggaatttt ctcgtgcccc aggtgcaacg aaacaaaatt tgacggccaa
     2881 cacaagcaac ataaactctt tgggtcttca aaacgatagt acatggtcaa ctggacgcag
     2941 tattggtgac ggttggcctg atccctcatc tgataacgag aataaagact ggtctgttgc
     3001 tcagccaact tcagcagcaa ctgcttacac tgatctggtc caagagtttg agccaggcaa
     3061 gccatggaag ggttcacaga tcaaaagcat agaagatgat cccagcatta caccaggaag
     3121 cgttgctaga tctccattgt ctattaattc gacgccaaaa gatgctgaca tatttgccaa
     3181 taccggtaaa aattcaccga ctgatttacc gccactaagt ttatcgtcgt ctacatggag
     3241 ttttaatcca aaccaaaatt atccgagtca cagttggtct gacaatagtc aacaatgtac
     3301 cgccacttcg gagctttgga caagcccgct aaataaatca tcgtctcgag gtcccccgcc
     3361 aggattgact gccaattcaa ataaatctgc aaatagtaat gcgtcaacgc caacaactat
     3421 taccggaggt gcgaatggat ggttacagcc tcgaagtggc ggtgttcaaa ccacaaacac
     3481 taattggaca ggtggtaaca ccacttgggg ctccagttgg ttgcttttga aaaatctaac
     3541 agcacagatt gatggtccta ctttgcgtac actgtgtatg cagcatgggc cccttgtcag
     3601 ctttcacccg tatttgaacc aaggaattgc cttatgtaaa tatactactc gtgaggaggc
     3661 gaacaaggcg caaatggcgt taaacaactg tgtcctcgcc aacaccacaa tatttgctga
     3721 atctcccagc gagaacgagg tgcaaagcat tatgcagcac ttaccacaaa ctccttcctc
     3781 tacaagctct agtggaacta gtggtggcaa cgtcggaggc gtcggcactt cagccaataa
     3841 tgcaaacagt ggttctgcag cttgtctgtc cggaaacaat agcggcaacg gaaacggcag
     3901 cgcgagcggc gccggcagcg gcaacaatgg caacagtagc tgcaacaaca gtgccgccgg
     3961 ggggggcagc agcagcaaca acacgattac cactgtagca aattcgaatc ttgttggttc
     4021 tagtggctct gtctcaaatt cctctggcgt tactgctaac tctagtactg tttctgtagt
     4081 tagttgtaca gcgagtggga attccataaa tggggcaggt actgcaaaca gttctggttc
     4141 aaagagtagt gcaaacaatt tagctagcgg ccagtctagc gcttctaact taactaatag
     4201 caccaattca acatggcgac aaactagcca aaaccaagct cttcaaagtc aaagcaggcc
     4261 atcaggcaga gaagctgact ttgattatat atctctcgtt tattccattg ttgatgatta
     4321 atctagaggg cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt
     4381 ctacgcgtac cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga
     4441 ctgtgccttc taaggcctga gctcgctgat cagcctcgat cgaggatcca gacatgataa
     4501 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     4561 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     4621 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     4681 aaagcaagta aaacctctac aaatgtggta tggctgatta tgatcagtcg accgatgccc
     4741 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     4801 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg
     4861 gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg
     4921 gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt
     4981 ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg
     5041 ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc
     5101 ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag
     5161 ggacagcttc aaggatcgct cgcggctctt accagccagc aaaaggccag gaaccgtaaa
     5221 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5281 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5341 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5401 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     5461 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5521 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5581 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5641 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     5701 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5761 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5821 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5881 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5941 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6001 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6061 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6121 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6181 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6241 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6301 gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6361 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6421 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6481 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6541 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6601 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6661 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6721 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6781 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6841 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6901 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6961 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     7021 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaaggaat taattctata
     7081 ttctaaaaac acaaatgata cttctaaaaa aaatcatgaa tggcatcaac tctgaatcaa
     7141 atctttgcag atgcacctac ttctcatttc cactgtcaca tcatttttcc agatctcgct
     7201 gcctgttatg tggcccacaa accaagacac gttttatggc cattaaagct ggctgatcgt
     7261 cgccaaacac caaatacata tcaatatgta cattcgagaa agaagcgatc aaagaagcgt
     7321 cttcgggcga gtaggagaat gcggaggaga aggagaacga gctgatctag tatctctcca
     7381 caatccaatg ccaactgacc aactggccat attcggagca atttgaagcc aatttccatc
     7441 gcctggcgat cgctccattc ttggctatat gtttttcacc gttcccgggg ccattttcaa
     7501 agactcgtcg gtaagataag attgtgtcac tcgctgtctc tcttcatttg tcgaagaatg
     7561 ctgaggaatt tcgcgatgac gtcggcgagt attttgaaga atgagaataa tttgtattta
     7621 tacgaaaatc agttagtgga attttctaca aaaaca
//
