LOCUS       sequence-288772-       10041 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmGW182-delUBA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10041)
  AUTHORS   Eulalio A, Helms S, Fritzsch C, Fauser M, Izaurralde E
  TITLE     A C-terminal silencing domain in GW182 is essential for miRNA 
            function.
  JOURNAL   RNA. 2009 Jun;15(6):1067-77. doi: 10.1261/rna.1605509. Epub 2009 Apr
            21.
  PUBMED    19383769
REFERENCE   2  (bases 1 to 10041)
  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: 
            "10.1261/rna.1605509"; journalName: "RNA"; date: "2009-06"; volume: 
            "15"; issue: "6"; pages: "1067-77"
COMMENT     Sequence Label: pAc5.1B-EGFP-DmGW182-delUBA_F
FEATURES             Location/Qualifiers
     source          1..10041
                     /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             7326..7367
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             7377..7394
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7417..7434)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7467..7601
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7491..7510)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7545..7564
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(8223..8811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8303..8322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8982..9773,9774..9842))
                     /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     9605..9624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9843..9947)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     10015..10033
                     /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 ggccgcatgc gtgaagccct
     3361 tttttcccaa gatggctggg gctgtcagca tgttaaccag gatactaatt gggaagttcc
     3421 cagttcgcca gaaccagcca ataaggatgc acccggtcca ccaatgtgga agccaagcat
     3481 taacaatggt actgatcttt gggagtccaa tttgagaaac ggaggtcagc cggccgcaca
     3541 gcaagttcca aagccgtcgt ggggtcatac accatcctct aacttaggtg gaacatgggg
     3601 tgaggacgac gatggcgccg atagtagtag tgtgtggact ggaggagctg ttagcaacgc
     3661 gggatccgga gctgcagtgg gagtaaacca agccggagtt aatgtcggtc caggcggtgt
     3721 tgtttcgtct ggcggacctc agtggggaca aggtgtcgtt ggcgtcggac ttggatcaac
     3781 tggaggtaac gggtcaagca atataactgg atcgtctgga gtcgcaacag gtagtagcgg
     3841 aaactccagc aacgctggta acggttgggg agaccctcgt gaaatacgcc ctttgggagt
     3901 tggtggctcc atggatattc gaaatgttga acatcgcggc ggtaacggtt ctggagcaac
     3961 ttcgagcgat ccacgagaca ttcgcatgat cgatccgcgt gaccctattc gaggagatcc
     4021 ccgtggaata tctggtcgtc ttaatgggac ctctgaaatg tggggtcatc atccacaaat
     4081 gtcccataac cagttgcaag gtatcaacaa aatggttggt caaagtgtag caactgccag
     4141 caccagtgtc ggaacatctg gctcgggcat cggtcctgga ggtcccggtc ctagtacagt
     4201 atcaggcaat atcccaacac agtgggggcc tgctcaaccg gtaagcgttg gtgtaagtgg
     4261 tcccaaagac atgtcaaaac agataagtgg atgggaggaa ccatcaccac cgcctcagcg
     4321 tcgcagtatt cctaactacg atgatggtac atcgttgtgg ggtcagcaaa ctcgtgttcc
     4381 cgctgcaagc ggtcactgga aagacatgac tgattcgata ggtcgtagta gtcatctcat
     4441 gcgtggccaa agccaaacgg gaggtatagg aatagccggc gttggaaata gcaatgttcc
     4501 agtgggagcc aatccaagta atcctataag cagtgtagtt ggacctcaag cccggattcc
     4561 atctgtgggc ggcgtacaac acaaaccaga cggcggcgct atgtgggtgc attccggcaa
     4621 tgtaggtggc agaaataatg ttgctgctgt tactacttgg ggagatgaca ctcatagcgt
     4681 taatgtcggc gctcccagca gtggcagtgt atccagcaac aattgggttg atgacaagtc
     4741 caactcaacc ttggcacaaa actcttggag cgacccggcc cctgttggag ttagttgggg
     4801 caataagcaa agcaaaccgc caagcaatag tgcttcatca ggttggagca ctgctgcggg
     4861 cgtggtggat ggggttgatc taggatctga gtggaacacg cacgggggga ttattggaaa
     4921 atctcagcaa caacaaaaac tagcgggact taacgtggga tcttatgccg accacaatag
     4981 ttcaacaagc agcggtggat ttgctggtcg ttacccggtc aacagtggac aaccttcaat
     5041 gtcctttcct cataataacc ttatgaataa catgggaggt accgctgtta ctggaggtaa
     5101 caacaataca aacatgacag ctttacaggt gcaaaagtat ttaaatcaag ggcaacatgg
     5161 tgtcgctgtt ggaccgcaag ccgttggtaa ttcttcagca gtatctgtcg gatttggtca
     5221 gaacacgtct aacgcagcag tggcaggagc agcctctgta aatatagcag caaatacaaa
     5281 caaccaaccg tctggtcagc aaattcgcat gctaggccag caaattcagt tggccattca
     5341 tagtggtttc atatctagtc agatattgac tcaaccgcta actcaaacaa cccttaacct
     5401 tttaaaccaa cttcttagca atattaagca tctccaggct gcgcagcaat cccttacccg
     5461 cgggggaaat gtcaatccaa tggcagtgaa tgtggctata tctaaataca agcagcaaat
     5521 ccagaattta cagaaccaga taaatgcaca acaggctgtg tatgtaaaac agcaaaatat
     5581 gcaaccaact tcacaacaac aacagcccca acaacagcaa cttccttctg ttcatctaag
     5641 taactcaggc aacgactatt taagaggtca cgatgcaata aataatttgc aaagcaactt
     5701 ttctgagctc aatattaata agccaagtgg atatcaagga gcgtccaatc aacaatcccg
     5761 attaaatcag tggaagcttc cagtattaga taaggagatc aactctgaca gtacggaatt
     5821 ttctcgtgcc ccaggtgcaa cgaaacaaaa tttgacggcc aacacaagca acataaactc
     5881 tttgggtctt caaaacgata gtacatggtc aactggacgc agtattggtg acggttggcc
     5941 tgatccctca tctgataacg agaataaaga ctggtctgtt gctcagccaa cttcagcagc
     6001 aactgcttac actgatctgg tccaagagtt tgagccaggc aagccatgga agggttcaca
     6061 gatcaaaagc atagaagatg atcccagcat tacaccagga agcgttgcta gatctccatt
     6121 gtctattaat tcgacgccaa aagatgctga catatttgcc aataccggta aaaattcacc
     6181 gactgattta ccgccactaa gtttatcgtc gtctacatgg agttttaatc caaaccaaaa
     6241 ttatccgagt cacagttggt ctgacaatag tcaacaatgt accgccactt cggagctttg
     6301 gacaagcccg ctaaataaat catcgtctcg aggtcccccg ccaggattga ctgccaattc
     6361 aaataaatct gcaaatagta atgcgtcaac gccaacaact attaccggag gtgcgaatgg
     6421 atggttacag cctcgaagtg gcggtgttca aaccacaaac actaattgga caggtggtaa
     6481 caccacttgg ggctccagtt ggttgctttt gaaaaatcta acagcacaga ttgatggtcc
     6541 tactttgcgt acactgtgta tgcagcatgg gccccttgtc agctttcacc cgtatttgaa
     6601 ccaaggaatt gccttatgta aatatactac tcgtgaggag gcgaacaagg cgcaaatggc
     6661 gttaaacaac tgtgtcctcg ccaacaccac aatatttgct gaatctccca gcgagaacga
     6721 ggtgcaaagc attatgcagc acttaccaca aactccttcc tctacaagct ctagtggaac
     6781 tagtggtggc aacgtcggag gcgtcggcac ttcagccaat aatgcaaaca gtggttctgc
     6841 agcttgtctg tccggaaaca atagcggcaa cggaaacggc agcgcgagcg gcgccggcag
     6901 cggcaacaat ggcaacagta gctgcaacaa cagtgccgcc ggggggggca gcagcagcaa
     6961 caacacgatt accactgtag caaattcgaa tcttgttggt tctagtggct ctgtctcaaa
     7021 ttcctctggc gttactgcta actctagtac tgtttctgta gttagttgta cagcgagtgg
     7081 gaattccata aatggggcag gtactgcaaa cagttctggt tcaaagagta gtgcaaacaa
     7141 tttagctagc ggccagtcta gcgcttctaa cttaactaat agcaccaatt caacatggcg
     7201 acaaactagc caaaaccaag ctcttcaaag tcaaagcagg ccatcaggca gagaagctga
     7261 ctttgattat atatctctcg tttattccat tgttgatgat taatctagag ggcccgcggt
     7321 tcgaaggtaa gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggtcatc
     7381 atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct tctaaggcct
     7441 gagctcgctg atcagcctcg atcgaggatc cagacatgat aagatacatt gatgagtttg
     7501 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     7561 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     7621 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     7681 acaaatgtgg tatggctgat tatgatcagt cgaccgatgc ccttgagagc cttcaaccca
     7741 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
     7801 tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac
     7861 cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac
     7921 gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc
     7981 attatcgccg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga
     8041 ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg
     8101 ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg
     8161 ctcgcggctc ttaccagcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     8221 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     8281 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     8341 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     8401 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     8461 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     8521 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     8581 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     8641 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     8701 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     8761 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     8821 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     8881 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     8941 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     9001 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     9061 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     9121 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     9181 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     9241 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca
     9301 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     9361 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     9421 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     9481 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     9541 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
     9601 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     9661 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     9721 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     9781 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     9841 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     9901 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     9961 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
    10021 cgtatcacga ggccctttcg t
//
