LOCUS       sequence-289508-        8430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmGW182_604-1384.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8430)
  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 8430)
  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_604-1384_N
FEATURES             Location/Qualifiers
     source          1..8430
                     /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             5715..5756
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5766..5783
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5806..5823)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5856..5990
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5880..5899)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5934..5953
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6612..7200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6692..6711)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7371..8162,8163..8231))
                     /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     7994..8013
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8232..8336)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8404..8422
                     /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 ggccgcggat cttatgccga
     3361 ccacaatagt tcaacaagca gcggtggatt tgctggtcgt tacccggtca acagtggaca
     3421 accttcaatg tcctttcctc ataataacct tatgaataac atgggaggta ccgctgttac
     3481 tggaggtaac aacaatacaa acatgacagc tttacaggtg caaaagtatt taaatcaagg
     3541 gcaacatggt gtcgctgttg gaccgcaagc cgttggtaat tcttcagcag tatctgtcgg
     3601 atttggtcag aacacgtcta acgcagcagt ggcaggagca gcctctgcaa atatagcagc
     3661 aaatacaaac aaccaaccgt ctggtcagca aattcgcatg ctaggccagc aaattcagtt
     3721 ggccattcat agtggtttca tatctagtca gatattgact caaccgctaa ctcaaacaac
     3781 ccttaacctt ttaaaccaac ttcttagcaa tattaagcat ctccaggctg cacagcaatc
     3841 ccttacccgc gggggaaatg tcaatccaat ggcagtgaat gtggctatat ctaaatacaa
     3901 gcagcaaatc cagaatttac agaaccagat aagtgcacaa caggctgtgt atgtaaaaca
     3961 gcaaaatatg caaccaactt cacaacaaca acagccccaa caacagcaac ttccttctgt
     4021 tcatctaagt aactcaggca acgactattt aagaggtcac gatgcaataa ataatttgca
     4081 aagcaacttt tctgagctca atattaataa gccaagtgga tatcaaggag cgtccaatca
     4141 acaatcccga ttaaatcagt ggaagcttcc agtattagat aaggagatca actctgacag
     4201 tacggaattt tctcgtgccc caggtgcaac gaaacaaaat ttgacggcca acacaagcaa
     4261 cataaactct ttgggtcttc aaaacgatag tacatggtca actggacgca gtattggtga
     4321 cggttggcct gatccctcat ctgataacga gaataaagac tggtctgttg ctcagccaac
     4381 ttcagcagca actgcttaca ctgatctggt ccaagagttt gagccaggca agccatggaa
     4441 gggttcacag atcaaaagca tagaagatga tcccagcatt acaccaggaa gcgttgctag
     4501 atctccattg tctattaatt cgacgccaaa agatgctgac atatttgcca ataccggtaa
     4561 aaattcaccg actgatttac cgccactaag tttatcgtcg tctacatgga gttttaatcc
     4621 aaaccaaaat tatccgagtc acagttggtc tgacaatagt caacaatgta ccgccacttc
     4681 ggagctttgg acaagcccgc taaataaatc atcgtctcga ggtcccccgc caggattgac
     4741 tgccaattca aataaatctg caaatagtaa tgcgtcaacg ccaacaacta ttaccggagg
     4801 tgcgaatgga tggttacagc ctcgaagtgg cggtgttcaa accacaaaca ctaattggac
     4861 aggtggtaac accacttggg gctccagttg gttgcttttg aaaaatctaa cagcacagat
     4921 tgatggtcct actttgcgta cactgtgtat gcagcatggg ccccttgtca gctttcaccc
     4981 gtatttgaac caaggaattg ccttatgtaa atatactact cgtgaggagg cgaacaaggc
     5041 gcaaatggcg ttaaacaact gtgtcctcgc caacaccaca atatttgctg aatctcccag
     5101 cgagaacgag gtgcaaagca ttatgcagca cttaccacaa actccttcct ctacaagctc
     5161 tagtggaact agtggtggca acgtcggagg cgtcggcact tcagccaata atgcaaacag
     5221 tggttctgca gcttgtctgt ccggaaacaa tagcggcaac ggaaacggca gcgcgagcgg
     5281 cgccggcagc ggcaacaatg gcaacagtag ctgcaacaac agtgccgccg gggggggcag
     5341 cagcagcaac aacacgatta ccactgtagc aaattcgaat cttgttggtt ctagtggctc
     5401 tgtctcaaat tcctctggcg ttactgctaa ctctagtact gtttctgtag ttagttgtac
     5461 agcgagtggg aattccataa atggggcagg tactgcaaac agttctggtt caaagagtag
     5521 tgcaaacaat ttagctagcg gccagtctag cgcttctaac ttaactaata gcaccaattc
     5581 aacatggcga caaactagcc aaaaccaagc tcttcaaagt caaagcaggc catcaggcag
     5641 agaagctgac tttgattata tatctctcgt ttattccatt gttgatgatt aatctagagg
     5701 gcccgcggtt cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta
     5761 ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt
     5821 ctaaggcctg agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg
     5881 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     5941 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     6001 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     6061 aaaacctcta caaatgtggt atggctgatt atgatcagtc gaccgatgcc cttgagagcc
     6121 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     6181 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc
     6241 ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga
     6301 atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag
     6361 aagcaggcca ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc
     6421 gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg
     6481 atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt
     6541 caaggatcgc tcgcggctct taccagccag caaaaggcca ggaaccgtaa aaaggccgcg
     6601 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6661 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6721 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6781 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     6841 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6901 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6961 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7021 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     7081 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7141 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7201 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7261 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7321 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7381 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7441 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7501 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7561 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7621 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7681 attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7741 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7801 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7861 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7921 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7981 gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8041 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8101 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8161 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8221 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8281 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8341 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     8401 aaaaataggc gtatcacgag gccctttcgt
//
