LOCUS       sequence-290291-        8496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmNot1_1-812.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8496)
  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 8496)
  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_1-812_W
FEATURES             Location/Qualifiers
     source          1..8496
                     /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             5781..5822
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5832..5849
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5872..5889)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5922..6056
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5946..5965)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6000..6019
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6678..7266)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6758..6777)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7437..8228,8229..8297))
                     /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     8060..8079
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8298..8402)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8470..8488
                     /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 tgaacgtaga gagccaactg aagacaccgc caacgcacat
     3361 tcgaaatctg gtcaagcacg tgaacaagcg aaattttaat gagagcagcg agcaaataaa
     3421 tcagttcgtt aaagagcacg gcctggaagc ggatcgcagc agcctgcgcc acctgttctc
     3481 cgtgatcaac ttcagcgatc tcgtgccatc agttacggtc cagctgcagg cgaagctcct
     3541 gggcatccag ttggagcgtc agcttcacag ttcatcgttt gtgtccaaca tctgctacgc
     3601 ttttgaccag ttcttcgcca gcaaccagaa gtccctgaag cctgttgcgg tagcggatct
     3661 cgttggccag gtcgcccgac tgaccggcat caacaaggtt tgtgagtgcg tctttgcctt
     3721 ggccttgacc cactcctcgt acacagaact caaacattct gccaggaaca acctaaaagg
     3781 tagcctcagc gaacttatag actcttactt gggtaataaa ggcaccggcc cagccgacag
     3841 cggtctccgg gagatctcct tcgatctact gcagtatttg ctgtgctgcc ttagcgagta
     3901 cgtacaaccg caggtggaag cacagttttt gatcaagctt cgtgaggagt ttccgcgcca
     3961 ggcggtaccg ctggttctag cgccgttcct ttacggctcg acgacggcga cgattgcggg
     4021 agcaggtgcc agcgaaacgg atgcggaggc cgaagcgacg actaatagca acagctccag
     4081 tttcgaagca gatgctttga acgaggtggg aatcgaggat atttatgacc atttaagcga
     4141 gataatattc accaaccagg gcaaaaataa tattatggac acatcttgga ttaatctaat
     4201 ccttgaaatc ggttatgaat ttacatcgag cgttgaagag tgtaagaatc atttgtgttc
     4261 ccgcgagcga gaacgcgctg agctccagtc caaagatgtc gccaagatcg tgggtcttat
     4321 gtgccgacgg cactcctccc tgctcgattg taatgtgaat ctaccgacgc cggcgaactt
     4381 ctggcctggt cagggacagg gcggtggcag caacagcagt ggctcttcgc agacacaaat
     4441 cactccgcag caacagaacc cgggcagcag taacaacaac gatggcagcg atggcaattc
     4501 gtccagtgat aagaaggaca agaaggagac gacggaggcc acgcaaactt ggaagccaga
     4561 tgtctttgtg caggcgctca aggaggtggt gcctcagctc aactggaagg acgtgtgcat
     4621 ggaacttgat catcccgagt tcgtgctgaa ggatcgcatc ggtttggagc ttttgctcac
     4681 catccttcgg ctggccactg gctctaacat atttccacat ccagaatgca tttaccgtca
     4741 ctgggcaaac acggaaggtc agctgtcgct tatcgcgaca atgctgaaga acccggatct
     4801 cttttccttc gccgactttg tatttagcca gccagcgttg gatgtgctta agacagctcc
     4861 ggatgcggac aataaggaga tctcgtcctg gaagtcgctt cacctggtgg aggtgctgct
     4921 ctccattgcg gacaagggat actatacgca ggtccatgag ctattcaagt tcccagcgca
     4981 gaactgtccc gatgtgctat ttttggcctt attgaacaca agtccgccta tgacgccact
     5041 gcgtcaggat ctgtttaacc aactgatacc aacgttcctt ggcaaccatc ccaactcgaa
     5101 cgtgatcttg gccagcgcct ggagctcgaa cagcttccaa cttcgttcca atatcatgaa
     5161 tgctatgtcc gagtggtatc tccgaggcaa tgagttcgat caggtgaaac tgtcgcgcat
     5221 cctggatctt gcccaggatt taaaagcgct atcagcgctg ctcaacgccc gttcattttt
     5281 attcattatc gacctggctt gccttgcctc tcggcgcgag tacctgaagc tggaaaagtg
     5341 gcttaccgat aaaattcgcg aacatggaga gccttttatg caggctataa tcaaggttct
     5401 gcaccgccgc tgcccgcagg ttatcaacgc caaagtgcca gaggatcagc tgcctcccaa
     5461 gcaggcgcag cttttgcccg agacagttac aaccatgatc aactgtctgc aaacgtgcat
     5521 caataactgc atgcagccgg agatggtcga agtcatcatg caaatgaccg cgaacgtggc
     5581 gattatggct aacatggcgc gtgcccaaca gcagcagcag ccaggattgg ttccgccgcc
     5641 tccaccaaca attctgcgcg gccatcgtgg tatggatctg cctggaggca tagtccctcc
     5701 gccaccgcag caaccgttct ccggaaacct caacgcgcag atgtttggac caggatgatc
     5761 tagagggccc gcggttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta
     5821 cgcgtaccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg
     5881 tgccttctaa ggcctgagct cgctgatcag cctcgatcga ggatccagac atgataagat
     5941 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     6001 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     6061 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     6121 gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcagtcgacc gatgcccttg
     6181 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca
     6241 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc
     6301 attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta
     6361 ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc
     6421 ggcgagaagc aggccattat cgccggcatg gcggccgacg cgctgggcta cgtcttgctg
     6481 gcgttcgcga cgcgaggctg gatggccttc cccattatga ttcttctcgc ttccggcggc
     6541 atcgggatgc ccgcgttgca ggccatgctg tccaggcagg tagatgacga ccatcaggga
     6601 cagcttcaag gatcgctcgc ggctcttacc agccagcaaa aggccaggaa ccgtaaaaag
     6661 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6721 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6781 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6841 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     6901 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6961 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7021 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7081 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     7141 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7201 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7261 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7321 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7381 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7441 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7501 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7561 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7621 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7681 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7741 gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7801 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7861 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7921 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7981 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     8041 tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8101 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8161 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8221 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8281 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8341 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8401 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     8461 acctataaaa ataggcgtat cacgaggccc tttcgt
//
