LOCUS       sequence-288766-       10131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmGW182-delmf1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10131)
  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 10131)
  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-delmf1_F
FEATURES             Location/Qualifiers
     source          1..10131
                     /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             7416..7457
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             7467..7484
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7507..7524)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7557..7691
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7581..7600)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7635..7654
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(8313..8901)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8393..8412)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9072..9863,9864..9932))
                     /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     9695..9714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9933..10037)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     10105..10123
                     /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 ggccgcggtc caccaatgtg
     3361 gaagccaagc attaacaatg gtactgatct ttgggagtcc aatttgagaa acggaggtca
     3421 gccggccgca cagcaagttc caaagccgtc gtggggtcat acaccatcct ctaacttagg
     3481 tggaacatgg ggtgaggacg acgatggcgc cgatagtagt agtgtgtgga ctggaggagc
     3541 tgttagcaac gcgggatccg gagctgcagt gggagtaaac caagccggag ttaatgtcgg
     3601 tccaggcggt gttgtttcgt ctggcggacc tcagtgggga caaggtgtcg ttggcgtcgg
     3661 acttggatca actggaggta acgggtcaag caatataact ggatcgtctg gagtcgcaac
     3721 aggtagtagc ggaaactcca gcaacgctgg taacggttgg ggagaccctc gtgaaatacg
     3781 ccctttggga gttggtggct ccatggatat tcgaaatgtt gaacatcgcg gcggtaacgg
     3841 ttctggagca acttcgagcg atccacgaga cattcgcatg atcgatccgc gtgaccctat
     3901 tcgaggagat ccccgtggaa tatctggtcg tcttaatggg acctctgaaa tgtggggtca
     3961 tcatccacaa atgtcccata accagttgca aggtatcaac aaaatggttg gtcaaagtgt
     4021 agcaactgcc agcaccagtg tcggaacatc tggctcgggc atcggtcctg gaggtcccgg
     4081 tcctagtaca gtatcaggca atatcccaac acagtggggg cctgctcaac cggtaagcgt
     4141 tggtgtaagt ggtcccaaag acatgtcaaa acagataagt ggatgggagg aaccatcacc
     4201 accgcctcag cgtcgcagta ttcctaacta cgatgatggt acatcgttgt ggggtcagca
     4261 aactcgtgtt cccgctgcaa gcggtcactg gaaagacatg actgattcga taggtcgtag
     4321 tagtcatctc atgcgtggcc aaagccaaac gggaggtata ggaatagccg gcgttggaaa
     4381 tagcaatgtt ccagtgggag ccaatccaag taatcctata agcagtgtag ttggacctca
     4441 agcccggatt ccatctgtgg gcggcgtaca acacaaacca gacggcggcg ctatgtgggt
     4501 gcattccggc aatgtaggtg gcagaaataa tgttgctgct gttactactt ggggagatga
     4561 cactcatagc gttaatgtcg gcgctcccag cagtggcagt gtatccagca acaattgggt
     4621 tgatgacaag tccaactcaa ccttggcaca aaactcttgg agcgacccgg cccctgttgg
     4681 agttagttgg ggcaataagc aaagcaaacc gccaagcaat agtgcttcat caggttggag
     4741 cactgctgcg ggcgtggtgg atggggttga tctaggatct gagtggaaca cgcacggggg
     4801 gattattgga aaatctcagc aacaacaaaa actagcggga cttaacgtgg gaatggtgaa
     4861 cgtaattaac gcggagatca ttaagcaaag caagcaatac aggatccttg tcgagaacgg
     4921 ctttaaaaag gaagatgtag agcgggcatt agtgattgct aatatgaaca tcgaagaggc
     4981 agccgatatg ctccgtgcca actcatccct atcaatggat ggttggcgtc gacatgatga
     5041 gtcccttgga tcttatgccg accacaatag ttcaacaagc agcggtggat ttgctggtcg
     5101 ttacccggtc aacagtggac aaccttcaat gtcctttcct cataataacc ttatgaataa
     5161 catgggaggt accgctgtta ctggaggtaa caacaataca aacatgacag ctttacaggt
     5221 gcaaaagtat ttaaatcaag ggcaacatgg tgtcgctgtt ggaccgcaag ccgttggtaa
     5281 ttcttcagca gtatctgtcg gatttggtca gaacacgtct aacgcagcag tggcaggagc
     5341 agcctctgta aatatagcag caaatacaaa caaccaaccg tctggtcagc aaattcgcat
     5401 gctaggccag caaattcagt tggccattca tagtggtttc atatctagtc agatattgac
     5461 tcaaccgcta actcaaacaa cccttaacct tttaaaccaa cttcttagca atattaagca
     5521 tctccaggct gcgcagcaat cccttacccg cgggggaaat gtcaatccaa tggcagtgaa
     5581 tgtggctata tctaaataca agcagcaaat ccagaattta cagaaccaga taaatgcaca
     5641 acaggctgtg tatgtaaaac agcaaaatat gcaaccaact tcacaacaac aacagcccca
     5701 acaacagcaa cttccttctg ttcatctaag taactcaggc aacgactatt taagaggtca
     5761 cgatgcaata aataatttgc aaagcaactt ttctgagctc aatattaata agccaagtgg
     5821 atatcaagga gcgtccaatc aacaatcccg attaaatcag tggaagcttc cagtattaga
     5881 taaggagatc aactctgaca gtacggaatt ttctcgtgcc ccaggtgcaa cgaaacaaaa
     5941 tttgacggcc aacacaagca acataaactc tttgggtctt caaaacgata gtacatggtc
     6001 aactggacgc agtattggtg acggttggcc tgatccctca tctgataacg agaataaaga
     6061 ctggtctgtt gctcagccaa cttcagcagc aactgcttac actgatctgg tccaagagtt
     6121 tgagccaggc aagccatgga agggttcaca gatcaaaagc atagaagatg atcccagcat
     6181 tacaccagga agcgttgcta gatctccatt gtctattaat tcgacgccaa aagatgctga
     6241 catatttgcc aataccggta aaaattcacc gactgattta ccgccactaa gtttatcgtc
     6301 gtctacatgg agttttaatc caaaccaaaa ttatccgagt cacagttggt ctgacaatag
     6361 tcaacaatgt accgccactt cggagctttg gacaagcccg ctaaataaat catcgtctcg
     6421 aggtcccccg ccaggattga ctgccaattc aaataaatct gcaaatagta atgcgtcaac
     6481 gccaacaact attaccggag gtgcgaatgg atggttacag cctcgaagtg gcggtgttca
     6541 aaccacaaac actaattgga caggtggtaa caccacttgg ggctccagtt ggttgctttt
     6601 gaaaaatcta acagcacaga ttgatggtcc tactttgcgt acactgtgta tgcagcatgg
     6661 gccccttgtc agctttcacc cgtatttgaa ccaaggaatt gccttatgta aatatactac
     6721 tcgtgaggag gcgaacaagg cgcaaatggc gttaaacaac tgtgtcctcg ccaacaccac
     6781 aatatttgct gaatctccca gcgagaacga ggtgcaaagc attatgcagc acttaccaca
     6841 aactccttcc tctacaagct ctagtggaac tagtggtggc aacgtcggag gcgtcggcac
     6901 ttcagccaat aatgcaaaca gtggttctgc agcttgtctg tccggaaaca atagcggcaa
     6961 cggaaacggc agcgcgagcg gcgccggcag cggcaacaat ggcaacagta gctgcaacaa
     7021 cagtgccgcc ggggggggca gcagcagcaa caacacgatt accactgtag caaattcgaa
     7081 tcttgttggt tctagtggct ctgtctcaaa ttcctctggc gttactgcta actctagtac
     7141 tgtttctgta gttagttgta cagcgagtgg gaattccata aatggggcag gtactgcaaa
     7201 cagttctggt tcaaagagta gtgcaaacaa tttagctagc ggccagtcta gcgcttctaa
     7261 cttaactaat agcaccaatt caacatggcg acaaactagc caaaaccaag ctcttcaaag
     7321 tcaaagcagg ccatcaggca gagaagctga ctttgattat atatctctcg tttattccat
     7381 tgttgatgat taatctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc
     7441 tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct
     7501 gatcagcctc gactgtgcct tctaaggcct gagctcgctg atcagcctcg atcgaggatc
     7561 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     7621 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     7681 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     7741 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt
     7801 cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg
     7861 actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg
     7921 gcagcgctct gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc
     7981 ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc
     8041 gccaccaaac gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg
     8101 ggctacgtct tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt
     8161 ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat
     8221 gacgaccatc agggacagct tcaaggatcg ctcgcggctc ttaccagcca gcaaaaggcc
     8281 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     8341 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     8401 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     8461 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     8521 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     8581 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     8641 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     8701 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     8761 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     8821 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     8881 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     8941 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     9001 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     9061 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     9121 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     9181 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     9241 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     9301 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     9361 agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt
     9421 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     9481 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     9541 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     9601 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     9661 cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact
     9721 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     9781 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     9841 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     9901 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     9961 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
    10021 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
    10081 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg t
//
