LOCUS       sequence-290293-        9465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmNot1_812-1945-K1537SG1562EQ1658A-dsRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9465)
  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 9465)
  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-K1537SG1562EQ1658A-dsRNAres_W
FEATURES             Location/Qualifiers
     source          1..9465
                     /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             6750..6791
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6801..6818
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6841..6858)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6891..7025
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6915..6934)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6969..6988
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7647..8235)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7727..7746)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8406..9197,9198..9266))
                     /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     9029..9048
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9267..9371)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9439..9457
                     /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 ttaattaaga tgggaatgga tccgctaacg aatatgtcca acaacttagc
     3361 cggcctcaac ctcagcggcc cgaacggagc atttaatttt ggaaacatgc taacttctcc
     3421 atcgcgtctg atgactccgg gagccaatcc ctatccgctg aacctcatgc agatgccgca
     3481 ggctccgcca ccacccaacg taggaaacct aggccgcatg ctgccagggg gtccacaaca
     3541 gcagactcca acgccgactc caacggcccc caatccaaac aaccctgtga tggccgatct
     3601 ccagatacct gtgtcaaagg aagttgaaga cgaggtcaac tcatactttc agcgcatcta
     3661 caaccaccag ccgaatccaa cgctgtccat cgatgaagtg ctggacattt tgcaaagatt
     3721 taaagagtca agtaaccggc gcgagcagga agtctttttg tgcatgctgc gcaatctgtt
     3781 cgaggaatat cgctttttct gtcaataccc agagaaagag ctacagatta ctgcgcagct
     3841 atttggtggc attattgatc gtaacctggt gcccactttt gttgcactgg gcctttctct
     3901 gcgctgtgtc ttggatgccc tccgcaagcc cgatggatct aagctttact atttcggtgt
     3961 gacggcactg gacagattca gaactcgatt gcacacctac aacaaatact gcgagcatat
     4021 ccgctccatt cctcacttct ctgacttccc gccgcacctt attcagtatg tggaatacgg
     4081 aatgcacggc caggagccgc cgccgcagaa gttgattgga ctcagcaata cgattccttc
     4141 ggcgatatct agcggaccag gaacggaacc aatttacagg aatagctcca tgttaggtaa
     4201 tatgcccgct gccaccccgg gatcgggacc gaaatcaaac gctgctgtat cgcatgccac
     4261 gagaatgaag tccatcgcca atgccaccaa tatcgacaca cttttggtgg ccaaccagga
     4321 agagaaagtg actgtgccac cagagcctgt gcaggacaaa actgccttta ttttcaacaa
     4381 cttgagccag ctgaacatac cgcagaagtg cgatgagata aaggagatta tgaccaagga
     4441 gtattggccg tggctagcgc aatatctggt cctcaaacgc gcatcaatgg agttcaactt
     4501 ccacacacta tattacaact ttttggatgc ccttaaaaac ggcgagatca accgattcgt
     4561 aaccaaagag accctacgca acatcaaggt actcttgcgc tccgataagg gagttattaa
     4621 tttctccgat cgaagtctgt tgaagaacct cggacactgg ttgggcatga tgaccttagg
     4681 tcgcaatcgc cccatcctgc agttggatct ggatcttaaa tcccttttgg ctgaggctta
     4741 ccacaagggc cagcaggaac tgctttttgt ggttcctttc gtagccaaga tccttgagtc
     4801 atccgccaag tctcgcatct tccgatcgcc caatccatgg acaatgggta tcatgtacgt
     4861 gcttgctgag cttcaccagg agccagacct taagctgaac ctgaagtttg aaatcgaagt
     4921 actctgcaaa acacttaatc tggagttggc caagttgcga caagtgatct acctgaagga
     4981 tcccaatcga actcaccgga ttgaagagca gatgtcgcaa ccaaaaccga aacaacttga
     5041 gccagtggct tctgcgccag ctctaccgcg tgaacagcaa tccccggcac aacctccgcc
     5101 accaccacaa cagcagcaac caccgcagca gcaggtacct ccaccgccat cctctgcgga
     5161 cgtggacgcc caaaatgcgg ccgctatgat gatggcagcg ggcggagcta atagtactcc
     5221 tggatcggta tcttcgccca atctacccac cgattccagc caggtggcac taccgccacc
     5281 agagccgcgc tattcatacg tggacgtaaa cgtgagcaac ttccagctta ttggtcagca
     5341 gttggtactg ccacccaata ctccgttcct gcacgccaat cccggaatca aacacatggt
     5401 cgtcaacgcc gtggagcgca cgataaccga ctggttgcag cctattgtgg accgaagcat
     5461 ccgcattgcc tgtgccacca ccgagcagat catccgcagc gacttcgccc tggatgctga
     5521 cgagaaccga atgcgcactg ccgctcatca gatggtacga aaccttgccg ccgagatggc
     5581 catgattacc ggtaaggatg agatagcgcg tgctattagc cagaatctgc acaaggcctt
     5641 gctgtcgggt ctaaatggga tgcccagcat ggctgagatc caagctgctg ctatgcagtt
     5701 agccagcgaa aatgttgagc tagtttgcgc tttcattcag aaaacatctg ctgaaaaggc
     5761 agccgcagag atcgataggc gtctaagcac cgatttcgag accagaaaaa ttgctcgtga
     5821 agagggcaac cgctttgtgg acgcccagat cctcacctat gcacaggagc gtcttccaga
     5881 agcagtgcgc atcaaggtgg gcgcggctcc agctacactt tatgctgtgt actcggagtt
     5941 cgctagaagc atacctggct tccagcagat gagcgatcgc gatattgccc tgtttgtgcc
     6001 caagccgacg gatttgtctc agccaaatgt gtttgctaat gatgatagca tggtatacgg
     6061 ggaactggct agcaaaatgg aggccttcat gaacacggca atcggagtgc cgacactcca
     6121 gatccaggct agcaagatgc acatgctact caacgccctg atagcgacgc gtcgactgcg
     6181 tgaccaagag tctgccttta atctgctgac ccgcgcggtc gagggtttga ccgaaggatt
     6241 ggtcaatatg catgaaaaca tggagcaaat gaagatgtac caaaacatcc atctccgtat
     6301 ccttggcttg ctgaacaaca gtttcggtgc tccaaacaca gagcgagcgg taacaaagtg
     6361 tttcttcgat atccgagagg aggtgcgata taacgtggag gcagctcgcg ctttgattac
     6421 gtcgcacttt gtcaatctga atcagtttga cggaatgcta cgcgactgca tggacaacgg
     6481 aaacaattat gtggctatat cgttcggtat cgcactgctg gagcgcctca tcatggacga
     6541 tcgagtgatt aatattgttt cggacaacga attcatggca actgttgagc tgctaggtag
     6601 gctgacccaa caccgtcacc gctaccccga gtgtatcgtc aacgccatcg acaccctgtg
     6661 gtccggcaat ttcaatagct cctccgatta ttcccccttt aatggaaacg accgctatct
     6721 gagctgatct agagggcccg cggttcgaag gtaagcctat ccctaaccct ctcctcggtc
     6781 tcgattctac gcgtaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag
     6841 cctcgactgt gccttctaag gcctgagctc gctgatcagc ctcgatcgag gatccagaca
     6901 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     6961 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     7021 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     7081 ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat cagtcgaccg
     7141 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc
     7201 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
     7261 ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg
     7321 cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc
     7381 aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac
     7441 gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct
     7501 tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac
     7561 catcagggac agcttcaagg atcgctcgcg gctcttacca gccagcaaaa ggccaggaac
     7621 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7681 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7741 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7801 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     7861 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7921 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7981 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     8041 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     8101 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     8161 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     8221 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     8281 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     8341 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     8401 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     8461 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8521 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8581 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8641 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8701 cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8761 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8821 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8881 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8941 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     9001 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     9061 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     9121 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     9181 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     9241 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     9301 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     9361 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     9421 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgt
//
