LOCUS       sequence-289509-        8337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmGW182_635-1384.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8337)
  AUTHORS   Huntzinger E, Kuzuoglu-Ozturk D, Braun JE, Eulalio A, Wohlbold L, 
            Izaurralde E
  TITLE     The interactions of GW182 proteins with PABP and deadenylases are 
            required for both translational repression and degradation of miRNA 
            targets.
  JOURNAL   Nucleic Acids Res. 2013 Jan;41(2):978-94. doi: 10.1093/nar/gks1078. 
            Epub 2012 Nov 21.
  PUBMED    23172285
REFERENCE   2  (bases 1 to 8337)
  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: 
            "10.1093/nar/gks1078"; journalName: "Nucleic Acids Res"; date: 
            "2013-01"; volume: "41"; issue: "2"; pages: "978-94"
COMMENT     Sequence Label: pAc5.1B-EGFP-DmGW182_635-1384_N
FEATURES             Location/Qualifiers
     source          1..8337
                     /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             5622..5663
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5673..5690
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5713..5730)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5763..5897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5787..5806)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5841..5860
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6519..7107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6599..6618)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7278..8069,8070..8138))
                     /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     7901..7920
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8139..8243)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8311..8329
                     /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 ggccgccata ataaccttat
     3361 gaataacatg ggaggtaccg ctgttactgg aggtaacaac aatacaaaca tgacagcttt
     3421 acaggtgcaa aagtatttaa atcaagggca acatggtgtc gctgttggac cgcaagccgt
     3481 tggtaattct tcagcagtat ctgtcggatt tggtcagaac acgtctaacg cagcagtggc
     3541 aggagcagcc tctgcaaata tagcagcaaa tacaaacaac caaccgtctg gtcagcaaat
     3601 tcgcatgcta ggccagcaaa ttcagttggc cattcatagt ggtttcatat ctagtcagat
     3661 attgactcaa ccgctaactc aaacaaccct taacctttta aaccaacttc ttagcaatat
     3721 taagcatctc caggctgcac agcaatccct tacccgcggg ggaaatgtca atccaatggc
     3781 agtgaatgtg gctatatcta aatacaagca gcaaatccag aatttacaga accagataag
     3841 tgcacaacag gctgtgtatg taaaacagca aaatatgcaa ccaacttcac aacaacaaca
     3901 gccccaacaa cagcaacttc cttctgttca tctaagtaac tcaggcaacg actatttaag
     3961 aggtcacgat gcaataaata atttgcaaag caacttttct gagctcaata ttaataagcc
     4021 aagtggatat caaggagcgt ccaatcaaca atcccgatta aatcagtgga agcttccagt
     4081 attagataag gagatcaact ctgacagtac ggaattttct cgtgccccag gtgcaacgaa
     4141 acaaaatttg acggccaaca caagcaacat aaactctttg ggtcttcaaa acgatagtac
     4201 atggtcaact ggacgcagta ttggtgacgg ttggcctgat ccctcatctg ataacgagaa
     4261 taaagactgg tctgttgctc agccaacttc agcagcaact gcttacactg atctggtcca
     4321 agagtttgag ccaggcaagc catggaaggg ttcacagatc aaaagcatag aagatgatcc
     4381 cagcattaca ccaggaagcg ttgctagatc tccattgtct attaattcga cgccaaaaga
     4441 tgctgacata tttgccaata ccggtaaaaa ttcaccgact gatttaccgc cactaagttt
     4501 atcgtcgtct acatggagtt ttaatccaaa ccaaaattat ccgagtcaca gttggtctga
     4561 caatagtcaa caatgtaccg ccacttcgga gctttggaca agcccgctaa ataaatcatc
     4621 gtctcgaggt cccccgccag gattgactgc caattcaaat aaatctgcaa atagtaatgc
     4681 gtcaacgcca acaactatta ccggaggtgc gaatggatgg ttacagcctc gaagtggcgg
     4741 tgttcaaacc acaaacacta attggacagg tggtaacacc acttggggct ccagttggtt
     4801 gcttttgaaa aatctaacag cacagattga tggtcctact ttgcgtacac tgtgtatgca
     4861 gcatgggccc cttgtcagct ttcacccgta tttgaaccaa ggaattgcct tatgtaaata
     4921 tactactcgt gaggaggcga acaaggcgca aatggcgtta aacaactgtg tcctcgccaa
     4981 caccacaata tttgctgaat ctcccagcga gaacgaggtg caaagcatta tgcagcactt
     5041 accacaaact ccttcctcta caagctctag tggaactagt ggtggcaacg tcggaggcgt
     5101 cggcacttca gccaataatg caaacagtgg ttctgcagct tgtctgtccg gaaacaatag
     5161 cggcaacgga aacggcagcg cgagcggcgc cggcagcggc aacaatggca acagtagctg
     5221 caacaacagt gccgccgggg ggggcagcag cagcaacaac acgattacca ctgtagcaaa
     5281 ttcgaatctt gttggttcta gtggctctgt ctcaaattcc tctggcgtta ctgctaactc
     5341 tagtactgtt tctgtagtta gttgtacagc gagtgggaat tccataaatg gggcaggtac
     5401 tgcaaacagt tctggttcaa agagtagtgc aaacaattta gctagcggcc agtctagcgc
     5461 ttctaactta actaatagca ccaattcaac atggcgacaa actagccaaa accaagctct
     5521 tcaaagtcaa agcaggccat caggcagaga agctgacttt gattatatat ctctcgttta
     5581 ttccattgtt gatgattaat ctagagggcc cgcggttcga aggtaagcct atccctaacc
     5641 ctctcctcgg tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa
     5701 cccgctgatc agcctcgact gtgccttcta aggcctgagc tcgctgatca gcctcgatcg
     5761 aggatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     5821 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     5881 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     5941 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg
     6001 atcagtcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     6061 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     6121 gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg
     6181 atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact
     6241 ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggccgac
     6301 gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt ccccattatg
     6361 attcttctcg cttccggcgg catcgggatg cccgcgttgc aggccatgct gtccaggcag
     6421 gtagatgacg accatcaggg acagcttcaa ggatcgctcg cggctcttac cagccagcaa
     6481 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6541 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6601 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6661 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     6721 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6781 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6841 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6901 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     6961 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7021 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7081 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7141 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7201 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7261 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7321 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7381 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7441 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7501 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7561 gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg
     7621 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7681 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7741 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7801 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7861 atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc
     7921 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7981 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8041 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8101 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8161 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8221 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     8281 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgt
//
