LOCUS       sequence-288927-        6978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmGW182-M2-Cterm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6978)
  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 6978)
  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-M2-Cterm_H
FEATURES             Location/Qualifiers
     source          1..6978
                     /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             3666..3707
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3717..3734
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3757..3774)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3807..3941
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3831..3850)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3885..3904
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4563..5151)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4643..4662)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5322..6113,6114..6182))
                     /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     5945..5964
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6183..6287)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6355..6373
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(6437..6978,1..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 ggcggccgcg ccaataccgg
     2761 taaaaattca ccgactgatt taccgccact aagtttatcg tcgtctacat ggagttttaa
     2821 tccaaaccaa aattatccga gtcacagttg gtctgacaat agtcaacaat gtaccgccac
     2881 ttcggagctt tggacaagcc cgctaaataa atcatcgtct cgaggtcccc cgccaggatt
     2941 gactgccaat tcaaataaat ctgcaaatag taatgcgtca acgccaacaa ctattaccgg
     3001 aggtgcgaat ggatggttac agcctcgaag tggcggtgtt caaaccacaa acactaattg
     3061 gacaggtggt aacaccactt ggggcccaca aactccttcc tctacaagct ctagtggaac
     3121 tagtggtggc aacgtcggag gcgtcggcac ttcagccaat aatgcaaaca gtggttctgc
     3181 agcttgtctg tccggaaaca atagcggcaa cggaaacggc agcgcgagcg gcgccggcag
     3241 cggcaacaat ggcaacagta gctgcaacaa cagtgccgcc ggggggggca gcagcagcaa
     3301 caacacgatt accactgtag caaattcgaa tcttgttggt tctagtggct ctgtctcaaa
     3361 ttcctctggc gttactgcta actctagtac tgtttctgta gttagttgta cagcgagtgg
     3421 gaattccata aatggggcag gtactgcaaa cagttctggt tcaaagagta gtgcaaacaa
     3481 tttagctagc ggccagtcta gcgcttctaa cttaactaat agcaccaatt caacatggcg
     3541 acaaactagc caaaaccaag ctcttcaaag tcaaagcagg ccatcaggca gagaagctga
     3601 ctttgattat atatctctcg tttattccat tgttgatgat taatctagag ggcccgcggt
     3661 tcgaaggtaa gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggtcatc
     3721 atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct tctaaggcct
     3781 gagctcgctg atcagcctcg atcgaggatc cagacatgat aagatacatt gatgagtttg
     3841 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     3901 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     3961 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     4021 acaaatgtgg tatggctgat tatgatcagt cgaccgatgc ccttgagagc cttcaaccca
     4081 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
     4141 tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac
     4201 cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac
     4261 gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc
     4321 attatcgccg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga
     4381 ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg
     4441 ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg
     4501 ctcgcggctc ttaccagcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4561 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4621 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4681 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4741 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4801 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4861 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4921 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4981 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     5041 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5101 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5161 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5221 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5281 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5341 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5401 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5461 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5521 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5581 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca
     5641 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5701 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5761 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5821 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5881 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
     5941 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6001 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6061 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6121 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6181 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6241 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6301 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     6361 cgtatcacga ggccctttcg tcttcaagga attaattcta tattctaaaa acacaaatga
     6421 tacttctaaa aaaaatcatg aatggcatca actctgaatc aaatctttgc agatgcacct
     6481 acttctcatt tccactgtca catcattttt ccagatctcg ctgcctgtta tgtggcccac
     6541 aaaccaagac acgttttatg gccattaaag ctggctgatc gtcgccaaac accaaataca
     6601 tatcaatatg tacattcgag aaagaagcga tcaaagaagc gtcttcgggc gagtaggaga
     6661 atgcggagga gaaggagaac gagctgatct agtatctctc cacaatccaa tgccaactga
     6721 ccaactggcc atattcggag caatttgaag ccaatttcca tcgcctggcg atcgctccat
     6781 tcttggctat atgtttttca ccgttcccgg ggccattttc aaagactcgt cggtaagata
     6841 agattgtgtc actcgctgtc tctcttcatt tgtcgaagaa tgctgaggaa tttcgcgatg
     6901 acgtcggcga gtattttgaa gaatgagaat aatttgtatt tatacgaaaa tcagttagtg
     6961 gaattttcta caaaaaca
//
