LOCUS       sequence-357560-        9441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmNot1_812-1945-dsRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9441)
  AUTHORS   Chen Y, Boland A, Kuzuoglu-Ozturk D, Bawankar P, Loh B, Chang CT, 
            Weichenrieder O, Izaurralde E
  TITLE     A DDX6-CNOT1 complex and W-binding pockets in CNOT9 reveal direct 
            links between miRNA target recognition and silencing.
  JOURNAL   Mol Cell. 2014 Jun 5;54(5):737-50. doi: 
            10.1016/j.molcel.2014.03.034. Epub 2014 Apr 24.
  PUBMED    24768540
REFERENCE   2  (bases 1 to 9441)
  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: 
            "10.1016/j.molcel.2014.03.034"; journalName: "Mol Cell"; date: 
            "2014-06-5- 5"; volume: "54"; issue: "5"; pages: "737-50"
COMMENT     Sequence Label: pAc5.1B-EGFP-DmNot1_812-1945-dsRNAres_W
FEATURES             Location/Qualifiers
     source          1..9441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2565
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2585..2587,2588..2590,2591..3301)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2585..2587,2588..2590,2591..3301)
                     /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(2630..2651)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2891..2910)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3238..3259
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6747..6788
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6798..6815
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6838..6855)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6888..7022
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6912..6931)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6966..6985
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7623..8211)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7703..7722)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8382..9173,9174..9242))
                     /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     9005..9024
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9243..9347)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9415..9433
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcaagaat taattctata ttctaaaaac acaaatgata cttctaaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat aatgaatatg tacacattcg
      241 agaaagaagc gatcaaagaa gcgtcttcgg gcggagtagg agaatgcgga ggagaaggag
      301 aacgagctga tctagtatct ctccacaatc caatgccaac tgaccaactg gccatattcg
      361 gagcaatttg aagccaattt ccatcgcctg gcgatcgctc cattcttggc tatatgtttt
      421 tcaccgttac ccggggccat tttcaaagac tcgtcggcaa gataagattg tgtcactcgc
      481 tgtctctctt catttgtcga agaatgctga ggaatttcgc gatgacgtcg gcgagtattt
      541 tgaagaatga gaataatttg tatttatacg aaaatcagtt agtggaattt tctacaaaaa
      601 catgttatct atagataatt ttgttgcaaa atatgttgac tatgacaaag attgtatgta
      661 tataccttta atgtattctc attttcttat gtatttataa tggcaatgat gatactgatg
      721 atattttaag atgatgccag accaaaaggc ttgaatttct gcgtcttttg ccgaacgcag
      781 tgcatgtgca attgttgttt tttggaatat tcaattttcg gactgtccgc tttgatttca
      841 gtttcttggc ttattcaaaa agcaaagtaa agccaaaaaa gcgagatggc aataccaaat
      901 gcggcaaaac ggtagtggaa ggaaaggggt gcggggcagc ggaaggaagg gtggggcggg
      961 gcgtggcggg gtctgtggct gggcgcgacg tcaccgacgt tggagccact cctttgacca
     1021 tgtgtgcgtg tgtgtattat tcgtgtctcg ccactcgccg gttgtttttt tctttttatg
     1081 ctgcgctctc tctagcgcca tctcgcttac gcatgctcaa cgcaccgcat gttgccgttt
     1141 ccttttatgc gtcattttgg ctcgaaatag gcaattattt aaacaaagat tagtcaacga
     1201 aaacgctaaa ataaataagt ctacaatatg gttacttatt gccatgtgtg tgcagccaac
     1261 gatagcaaca aaagcaacaa cacaggtggc tttccctctt tcactttttg tttgcaagcc
     1321 gcgtgcgagc aagacggcac gaccggcaaa cgcaattacg ctgacaaaga gcagacgaag
     1381 ttttggcgaa aaacatcaag gcgcctgata cgaatgcatt tgcaataaca attgcgatat
     1441 ttaatattgt ttatgaagct gtttgacttc aaaacacaca aaaaaaaaaa taaaacaaat
     1501 tatttgaaag agaattagga atcggacgct tatcgttagg gtaacaacaa gaaatgctta
     1561 ctgagtcaca gcctctggaa aactgccgca agccagagag agagagaaaa agagggagag
     1621 cagcttagac cgcatgtgct tgtgtgtgag gcgtctctct cttcgtctct gttgcgcaaa
     1681 cgcatagact gcactgaaaa aatcgattac ctatttttta tgaatgaata tttgcactat
     1741 tactattcaa aactattaag atagcaatca cattcaatag ccaaatacta taccacctga
     1801 gcgatgcaac gaaatgatca atttgagcaa aaatgctgca tatttaggac ggcatcatta
     1861 tagaaatgct tcttgctgtg tacttttctc tcgtctggca gctgtttcgc cgttattgtt
     1921 aaaaccggct taagttaggt gtgttttcta cgactagtga atgccctact agaagatgtg
     1981 tgttgcacaa aatgtccctg gaataaccaa tttgaagtgc agatagcagt aaacgtaagc
     2041 taatatgaat attatttaac tgtaatgttt taatatcgct ggacattact aataaaccca
     2101 ctataaacac atgtacatat gtatgttttg gcatacaatg agtagttggg gaaaaaatgt
     2161 gtaaaagcac cgtgaccatc acagcataaa gataaccagc tgaagtatcg aatatgagta
     2221 acccccaaat tgaatcacat gccgcaactg ataggaccca tggaagtaca ctcttcatgg
     2281 cgatatacaa gacacacaca agcacgaaca cccagttgcg gaggaaattc tccgtaaatg
     2341 aaaacccaat cggcgaacaa ttcataccca tatatggtaa aagttttgaa cgcgacttga
     2401 gagcggagag cattgcggct gataaggttt tagcgctaag cgggctttat aaaacgggct
     2461 gcgggaccag ttttcatatc actaccgttt gagttcttgt gctgtgtgga tactcctccc
     2521 gacacaaagc cgctccatca gccagcagtc gtctaatcca gagaccccgg atcggggtac
     2581 catcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2641 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     2701 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2761 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2821 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2881 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2941 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     3001 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     3061 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     3121 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3181 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     3241 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     3301 gccaagctta attaagatgg gaatggatcc gctaacgaat atgtccaaca acttagccgg
     3361 cctcaacctc agcggcccga acggagcatt taattttgga aacatgctaa cttctccatc
     3421 gcgtctgatg actccgggag ccaatcccta tccgctgaac ctcatgcaga tgccgcaggc
     3481 tccgccacca cccaacgtag gaaacctagg ccgcatgctg ccagggggtc cacaacagca
     3541 gactccaacg ccgactccaa cggcccccaa tccaaacaac cctgtgatgg ccgatctcca
     3601 gatacctgtg tcaaaggaag ttgaagacga ggtcaactca tactttcagc gcatctacaa
     3661 ccaccagccg aatccaacgc tgtccatcga tgaagtgctg gacattttgc aaagatttaa
     3721 agagtcaagt aaccggcgcg agcaggaagt ctttttgtgc atgctgcgca atctgttcga
     3781 ggaatatcgc tttttctgtc aatacccaga gaaagagcta cagattactg cgcagctatt
     3841 tggtggcatt attgatcgta acctggtgcc cacttttgtt gcactgggcc tttctctgcg
     3901 ctgtgtcttg gatgccctcc gcaagcccga tggatctaag ctttactatt tcggtgtgac
     3961 ggcactggac agattcagaa ctcgattgca cacctacaac aaatactgcg agcatatccg
     4021 ctccattcct cacttctctg acttcccgcc gcaccttatt cagtatgtgg aatacggaat
     4081 gcacggccag gagccgccgc cgcagaagtt gattggactc agcaatacga ttccttcggc
     4141 gatatctagc ggaccaggaa cggaaccaat ttacaggaat agctccatgt taggtaatat
     4201 gcccgctgcc accccgggat cgggaccgaa atcaaacgct gctgtatcgc atgccacgag
     4261 aatgaagtcc atcgccaatg ccaccaatat cgacacactt ttggtggcca accaggaaga
     4321 gaaagtgact gtgccaccag agcctgtgca ggacaaaact gcctttattt tcaacaactt
     4381 gagccagctg aacataccgc agaagtgcga tgagataaag gagattatga ccaaggagta
     4441 ttggccgtgg ctagcgcaat atctggtcct caaacgcgca tcaatggagt tcaacttcca
     4501 cacactatat tacaactttt tggatgccct taaaaacggc gagatcaacc gattcgtaac
     4561 caaagagacc ctacgcaaca tcaaggtact cttgcgctcc gataagggag ttattaattt
     4621 ctccgatcga agtctgttga agaacctcgg acactggttg ggcatgatga ccttaggtcg
     4681 caatcgcccc atcctgcagt tggatctgga tcttaaatcc cttttggctg aggcttacca
     4741 caagggccag caggaactgc tttttgtggt tcctttcgta gccaagatcc ttgagtcatc
     4801 cgccaagtct cgcatcttcc gatcgcccaa tccatggaca atgggtatca tgtacgtgct
     4861 tgctgagctt caccaggagc cagaccttaa gctgaacctg aagtttgaaa tcgaagtact
     4921 ctgcaaaaca cttaatctgg agttggccaa gttgcgacaa gtgatctacc tgaaggatcc
     4981 caatcgaact caccggattg aagagcagat gtcgcaacca aaaccgaaac aacttgagcc
     5041 agtggcttct gcgccagctc taccgcgtga acagcaatcc ccggcacaac ctccgccacc
     5101 accacaacag cagcaaccac cgcagcagca ggtacctcca ccgccatcct ctgcggacgt
     5161 ggacgcccaa aatgcggccg ctatgatgat ggcagcgggc ggagctaata gtactcctgg
     5221 atcggtatct tcgcccaatc tacccaccga ttccagccag gtggcactac cgccaccaga
     5281 gccgcgctat tcatacgtgg acgtaaacgt gagcaacttc cagcttattg gtcagcagtt
     5341 ggtactgcca cccaatactc cgttcctgca cgccaatccc ggaatcaaac acatggtcgt
     5401 caacgccgtg gagcgcacga taaccgactg gttgcagcct attgtggacc gaagcatccg
     5461 cattgcctgt gccaccaccg agcagatcat ccgcaaggac ttcgccctgg atgctgacga
     5521 gaaccgaatg cgcactgccg ctcatcagat ggtacgaaac cttgccgccg gcatggccat
     5581 gattaccggt aaggatgaga tagcgcgtgc tattagccag aatctgcaca aggccttgct
     5641 gtcgggtcta aatgggatgc ccagcatggc tgagatccaa gctgctgcta tgcagttagc
     5701 cagcgaaaat gttgagctag tttgcgcttt cattcagaaa acatctgctg aaaaggcagc
     5761 cgcagagatc gataggcgtc taagcaccga tttcgagacc agaaaaattg ctcgtgaaga
     5821 gggcaaccgc tttgtggacg cccagatcct cacctatcaa caggagcgtc ttccagaagc
     5881 agtgcgcatc aaggtgggcg cggctccagc tacactttat gctgtgtact cggagttcgc
     5941 tagaagcata cctggcttcc agcagatgag cgatcgcgat attgccctgt ttgtgcccaa
     6001 gccgacggat ttgtctcagc caaatgtgtt tgctaatgat gatagcatgg tatacgggga
     6061 actggctagc aaaatggagg ccttcatgaa cacggcaatc ggagtgccga cactccagat
     6121 ccaggctagc aagatgcaca tgctactcaa cgccctgata gcgacgcgtc gactgcgtga
     6181 ccaagagtct gcctttaatc tgctgacccg cgcggtcgag ggtttgaccg aaggattggt
     6241 caatatgcat gaaaacatgg agcaaatgaa gatgtaccaa aacatccatc tccgtatcct
     6301 tggcttgctg aacaacagtt tcggtgctcc aaacacagag cgagcggtaa caaagtgttt
     6361 cttcgatatc cgagaggagg tgcgatataa cgtggaggca gctcgcgctt tgattacgtc
     6421 gcactttgtc aatctgaatc agtttgacgg aatgctacgc gactgcatgg acaacggaaa
     6481 caattatgtg gctatatcgt tcggtatcgc actgctggag cgcctcatca tggacgatcg
     6541 agtgattaat attgtttcgg acaacgaatt catggcaact gttgagctgc taggtaggct
     6601 gacccaacac cgtcaccgct accccgagtg tatcgtcaac gccatcgaca ccctgtggtc
     6661 cggcaatttc aatagctcct ccgattattc cccctttaat ggaaacgacc gctatctgag
     6721 ctgatctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg
     6781 attctacgcg taccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct
     6841 cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga
     6901 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     6961 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     7021 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     7081 tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgaccgatg
     7141 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
     7201 gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc
     7261 tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt
     7321 gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa
     7381 cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc
     7441 ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct tctcgcttcc
     7501 ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat
     7561 cagggacagc ttcaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     7621 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     7681 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     7741 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     7801 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7861 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7921 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7981 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     8041 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     8101 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     8161 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     8221 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     8281 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     8341 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     8401 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     8461 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     8521 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     8581 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     8641 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca
     8701 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     8761 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     8821 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     8881 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     8941 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
     9001 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     9061 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     9121 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     9181 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     9241 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     9301 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     9361 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     9421 cgtatcacga ggccctttcg t
//
