LOCUS       sequence-289147-        8451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmNot1_1710-2505.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8451)
  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 8451)
  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_1710-2505_J
FEATURES             Location/Qualifiers
     source          1..8451
                     /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             5736..5777
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5787..5804
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5827..5844)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5877..6011
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5901..5920)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5955..5974
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6633..7221)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6713..6732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7392..8183,8184..8252))
                     /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     8015..8034
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8253..8357)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8425..8443
                     /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 tgtctcagcc aaatgtgttt gctaatgatg atagcatggt
     3361 atacggggaa ctggctagca aaatggaggc cttcatgaac acggcaatcg gagtgccgac
     3421 actccagatc cagactagca agatgcacat gctactcaac gccctgatag cgacgcgtcg
     3481 actgcgtgac caagagtctg cctttaatct gctgacccgc gcggtcgagg gtttgaccga
     3541 aggattggtc aatatgcatg aaaacatgga gcaaatgaag atgtaccaaa acatccatct
     3601 ccgtatcctt ggcttgctga acaacagttt cggtgctcca aacacagagc gagcggtaac
     3661 aaagtgtttc ttcgatatcc gagaggaggt gcgatataac gtggaggcag ctcgcgcttt
     3721 gattacgtcg cactttgtca atctgaatca gtttgacgga atgctacgcg actgcatgga
     3781 caacggaaac aattatgtgg ccatatcgtt cggtatcgca ctgctggagc gcctcatcat
     3841 ggacgatcga gtgattaata ttgtttcgga caacgaattc atggcaactg ttgagctgct
     3901 aggtaggctc actcagcatc gccatcggta tccagaatgc attgtgaatg ctattgatac
     3961 gctctggagc ggaaacttta actcgagcag cgactacagc ccgttcaacg gcaatgatcg
     4021 ttacttgtcg ggagcctccc attacatcca ctccggcatg catcacgtaa ggtcatgcga
     4081 tacagatgat ccgccgggct tacaagagaa gacggagttt ctgcttaagg attgggttgc
     4141 actgtacacg caacagaacc agcagtcaac gcgcgatgca cgcaacttcg gtgcatttgt
     4201 tcagaagatg aacacatatg gtatcctgaa aacggacgac ttgatcaccc gttttttccg
     4261 ccaggccaca cacatttgca cggatgtggt gtacagaatg ttcgccgagc cgagtttacc
     4321 aatcaaccag gccaaaaaca agatattcca gtggatcgat gcgttcgtcc atctgattgc
     4381 aatgctcgtg cgacactcag gcgaagcagg taatccaacc accaagatca acctgttaaa
     4441 caaggtgctg ggcattgtgc tgggaacgct gattaaggac catgagatgc gtggcgttag
     4501 cttccagcag gttggctacc atcgcttctt catgatgctg ttcatggagc tgtgcacggc
     4561 cgatgtgatt ctcgagtccc tgatgcacag tattgtgtcg gccttcgcct acacatatca
     4621 cttgttgaat cccagcgtgg ctccgggatt ctgctttgct tggttggagc tcatttcgca
     4681 tcgcgtcttc ctgggccgca tcctggtcca aattcccggg caaaagggct ggccacttta
     4741 cgctcagttg ctgcaggacc tttttaaata tcttgcgccc ttcttacgca acactgagct
     4801 cggcaaacca gtccagcttt tgtacaaggg gacgctacgt gttctgctcg ttctgcttca
     4861 cgacttcccg gagttcttgt gcgactatca ttttgggttc tgtgacacca tcccgcccaa
     4921 ttgcgtccag atgcgtaaca tcatcctgtc ggcgtttccg cgcaacatgc gcttgcccga
     4981 tcccttcacg cccaacctaa aagtggacat gctgtcggac agtagtaatg cgcctaaggt
     5041 gctcagtagc tacattatga acatccagcc cgcgaacttt aagaaggatt tggactcgta
     5101 cctcaaggcc cgggcgccag ttacttttct atcggaatta cgtggtcact tgcaggtgac
     5161 aagcgagcca ggaactcgtt acaacatggc actgatgaat gctctggtca tgtatgtggg
     5221 aactcaggcg attgctttga tcagaaacaa aaactttgtg cccaacacct cgaacatagc
     5281 acacagcgcc cacatggata tcttccagaa tcttgctgtt gacttggata cggagggtcg
     5341 ctatctgttc ttgaatgcga ttgcgaacca gttgcgctat cccaacagcc acacgcacta
     5401 cttcagctgt gcggtgctgc atctgtttgc cgaggccaac tcggaggcaa ttcaggagca
     5461 gatcactcgc gtacttctgg aacgattgat cgtgaaccgc ccgcatcctt ggggactgtt
     5521 gatcactttt attgagctga tcaaaaatcc catctacaag ttctgggacc atgactttgt
     5581 gcactgtgcg ccagagatca ccaagctctt tgaatcggtg gcccgttcct gtctggccaa
     5641 gtcgaacgtc acccagcagt tgaacatgcc cgtcgtcgat ggcgagggcc aggaggtagc
     5701 caccatcaac tgatctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc
     5761 tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct
     5821 gatcagcctc gactgtgcct tctaaggcct gagctcgctg atcagcctcg atcgaggatc
     5881 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     5941 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     6001 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     6061 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt
     6121 cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg
     6181 actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg
     6241 gcagcgctct gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc
     6301 ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc
     6361 gccaccaaac gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg
     6421 ggctacgtct tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt
     6481 ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat
     6541 gacgaccatc agggacagct tcaaggatcg ctcgcggctc ttaccagcca gcaaaaggcc
     6601 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6661 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6721 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6781 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     6841 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6901 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6961 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7021 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     7081 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7141 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7201 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7261 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7321 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7381 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7441 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7501 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7561 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7621 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7681 agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt
     7741 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7801 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7861 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7921 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7981 cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact
     8041 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8101 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8161 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8221 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8281 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8341 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     8401 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg t
//
