LOCUS       sequence-288764-        9025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmAGO1-PAZ6mut-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9025)
  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 9025)
  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-DmAGO1-PAZ6mut-siRNAres_F
FEATURES             Location/Qualifiers
     source          1..9025
                     /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             6310..6351
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6361..6378
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6401..6418)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6451..6585
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6475..6494)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6529..6548
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7207..7795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7287..7306)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7966..8757,8758..8826))
                     /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     8589..8608
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8827..8931)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8999..9017
                     /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 ttgaattcta tgtccacgga gcgtgagctg gctcccggtg ggccagctca
     3361 gctccatccg cacacgctgc cgctgacttt tccggatctc cagatgacct ccaccgtggg
     3421 catcattggg aaagtctacg agtcacagtg gaccccctcg cccactcggc ctcagagtcc
     3481 ctctcaggcg cagactagct tcgacacgct cacatcacca ccagcgcccg gctcgtcggt
     3541 caatcccacc gccgtcacca gcccaagtgc ccagaatgtg gccgctggtg gagcaactgt
     3601 ggccggtgca gcagcaactg ccgcccaggt ggcctccgcc ttgggtgcca ccaccggcag
     3661 cgtgacgcca gcaattgcca ccgcaacgcc agccacgcag ccggatatgc ccgtctttac
     3721 gtgtccacga cgcccgaatc tcggacgaga gggtcgcccg attgtgctgc gcgccaatca
     3781 cttccaggtg acaatgccgc gtggctatgt gcatcactat gacatcaata tacagccgga
     3841 caagtgtccg cgaaaggtga accgtgagat tatcgagact atggtgcatg cctatagcaa
     3901 gatcttcgga gtgctcaagc cggtgttcga tggtcgcaac aatctgtata cccgcgatcc
     3961 cctgcccatt ggcaacgagc gtctagagct ggaggttact ctacccggcg agggcaaaga
     4021 tcgaatcttt cgcgtgacga tcaagtggca ggctcaggtc tcgctcttca atttggagga
     4081 agctctcgaa ggccgcacgc ggcagatacc ctatgatgcc attttggcgc tcgatgtggt
     4141 catgcgccat ctgcccagca tgacgtacac gccagtggga cgtagcttct tcagttcccc
     4201 ggagggttac taccatcccc tgggtggtgg acgcgaggtt tggttcggtt tccatcagag
     4261 cgcaaggccc tcgcagtgga agatgatgct caatatcgat gtctcggcca ccgctttcta
     4321 caaggctcaa ccagtcattg acttcatgtg cgaggtgctg gacattcgcg acatcaacga
     4381 gcagcgcaaa ccgctcaccg attcgcagcg cgtcaagttc accaaagaaa ttaaaggcct
     4441 gaagatcgag atcacccact gcggccagat gcgtgccgcg gctcgtgtgt gcaacgtcac
     4501 tcgccgcccc gctcagatgc aatcattccc actgcagctg gagaacggac agaccgtaga
     4561 gtgcaccgtg gccaagtact tcctggacaa gtaccgcatg aaattgcgct acccgcactt
     4621 gccctgcctg caggttggcc aagagcacaa gcacgcggcc gcgcctctag aggtgtgcaa
     4681 cattgtggcc ggacagcggt gcattaaaaa gctgaccgat atgcagacgt cgaccatgat
     4741 caaggccaca gctcgttcag ctccggatcg tgagcgtgag attaacaact tggtaaagcg
     4801 cgccgacttc aacaacgatt cgtatgtgca agagtttggc ctgaccatct ccaattcgat
     4861 gatggaggta cgaggacgcg tcttgcctcc tcccaagctt cagtatgggg gacgtgtgtc
     4921 taccggcctc accggccagc agctgttccc gccacagaac aaggtgagct tggcctcgcc
     4981 caaccagggt gtatgggata tgcgaggcaa gcagttcttc actggcgtcg agatccgcat
     5041 ctgggccatc gcctgttttg ccccacagcg cacggtgcgc gaggatgcgc tgcgtaattt
     5101 cacccagcag ctgcagaaga tctcaaacga tgcaggcatg ccgataattg gacagccgtg
     5161 cttctgtaag tacgccaccg ggccggatca agtggaaccc atgttccgtt acctgaagat
     5221 caccttcccc ggcctgcaac tcgtcgtggt tgtgctgccc ggcaagactc cagtatacgc
     5281 cgaggtgaag cgtgtaggtg acaccgttct gggtatggcc acccagtgtg tgcaggccaa
     5341 gaacgtgaac aagacatcgc cacagacgct ctctaatctg tgtctgaaga tcaacgtcaa
     5401 gttgggcggc atcaattcaa ttctggtccc ctccattcgg ccaaaggtct tcaatgagcc
     5461 ggtcatcttt ttgggtgccg atgtgacaca cccaccagct ggcgacaaca agaaaccatc
     5521 gattgccgcc gtcgtgggct ccatggatgc ccatccatcg cggtatgccg ccaccgttcg
     5581 cgtacagcag caccgacagg agatcatcca ggagctgagc agcatggtgc gcgagctgtt
     5641 gatcatgttc tacaagtcga cgggcggcta caagccccac cgcatcatac tctatcgtga
     5701 cggagtctcc gagggacaat tcccacatgt cctgcaacac gaattgaccg ccattcgcga
     5761 ggcctgcatt aagctagagc cagagtatcg gccgggcatc acattcattg tggtgcagaa
     5821 gcgccatcac actcgactct tctgcgcgga gaagaaggag cagagcggca aatcgggcaa
     5881 tattcccgca ggcaccaccg tcgatgtggg catcacacat cccaccgaat ttgatttcta
     5941 tctatgcagc catcagggca tccagggcac cagtcgcccc tcgcactacc acgttctgtg
     6001 ggacgacaat cactttgact cggacgagct gcagtgcctc acgtatcagc tatgccgtac
     6061 gtacgtgcgc tgcacccgat ccgtcagtat accggcgcca gcctactacg cccatttagt
     6121 ggccttccgt gccagatatc atctggtgga gaaggagcac gattcgggcg agggttcgca
     6181 ccagagcggc tgctcagagg atcgtacgcc aggtgccatg gccagggcca tcactgtgca
     6241 cgcggatacc aagaaggtca tgtactttgc ctaagcggcc gctcgagtct agagggcccg
     6301 cggttcgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac gcgtaccggt
     6361 catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctaag
     6421 gcctgagctc gctgatcagc ctcgatcgag gatccagaca tgataagata cattgatgag
     6481 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     6541 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     6601 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     6661 ctctacaaat gtggtatggc tgattatgat cagtcgaccg atgcccttga gagccttcaa
     6721 cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
     6781 cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca ttttcggcga
     6841 ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt
     6901 gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca
     6961 ggccattatc gccggcatgg cggccgacgc gctgggctac gtcttgctgg cgttcgcgac
     7021 gcgaggctgg atggccttcc ccattatgat tcttctcgct tccggcggca tcgggatgcc
     7081 cgcgttgcag gccatgctgt ccaggcaggt agatgacgac catcagggac agcttcaagg
     7141 atcgctcgcg gctcttacca gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7201 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7261 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7321 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7381 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7441 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7501 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7561 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7621 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     7681 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7741 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7801 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7861 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7921 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7981 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     8041 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     8101 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     8161 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8221 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8281 tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8341 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8401 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8461 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8521 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8581 aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8641 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8701 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8761 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8821 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8881 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8941 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     9001 taggcgtatc acgaggccct ttcgt
//
