LOCUS       sequence-289786-        9221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmNot4-dsRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9221)
  AUTHORS   Keskeny C, Raisch T, Sgromo A, Igreja C, Bhandari D, Weichenrieder 
            O, Izaurralde E
  TITLE     A conserved CAF40-binding motif in metazoan NOT4 mediates 
            association with the CCR4-NOT complex.
  JOURNAL   Genes Dev. 2019 Feb 1;33(3-4):236-252. doi: 10.1101/gad.320952.118. 
            Epub 2019 Jan 28.
  PUBMED    30692204
REFERENCE   2  (bases 1 to 9221)
  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"; journalName: "Genes 
            Dev."; date: "2019-02-1"; volume: "33(3-4):236-252. doi"; pages: " 
            10.1101/gad.320952.118. Epub 2019 Jan 28"
COMMENT     Sequence Label: pAc5.1B-EGFP-DmNot4-dsRNAres_Q
FEATURES             Location/Qualifiers
     source          1..9221
                     /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             6506..6547
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6557..6574
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6597..6614)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6647..6781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6671..6690)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6725..6744
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7403..7991)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7483..7502)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8162..8953,8954..9022))
                     /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     8785..8804
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9023..9127)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9195..9213
                     /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 ttgaattctg cagatatcca gcacagtggc ggccgctcga gtatgaacgg
     3361 cctgagcagc aatgatgatg cagtcgagtg ccctttgtgt atggagccgc ttgaggtgga
     3421 cgatctgacc ttctttccgt gtacctgcgg atatcagatt tgccgattct gttggcatag
     3481 gatccgcact gatgagaaca agctgtgtcc ggcatgccga aaggagtatc ccgagaatcc
     3541 agctgacttt aagccgctgt cgcaggagga aatgattgcc ttcaagtccc agaaacgcca
     3601 aagggaccaa caacgaaaac aaaagatcac cgagaaccgc aaacacttgg ccaacgttcg
     3661 cgttgtccaa aagaacttgg tgttcgttgt ggggctacca ccacgacttg ctgatgcaga
     3721 catattaaag aaacacgagt atttcggtaa atatgggaaa attcataaag tcgttataaa
     3781 tccaagtacc acgtatgccg gggtccaggg tccatctgct tctgcgtatg tcacatatgt
     3841 taataactcg gatgccttgc gggccattca aagcgtcaat aatattatga tagacggtcg
     3901 gcttataaag accagcttgg ggacaaccaa atactgtagc cacttcatga agaaccagca
     3961 gtgtcccaag agcgactgca tgtacttaca tgaattgggc gatcccgagg ccagtttcac
     4021 aaaggaggaa atgcatcagg gaaagcacct ggagtacgag aagcgcctgc acgatactct
     4081 aattgcgtca ttgggaccga atgcaacagg tattgtaaat gtcgcgattc catcgtcatc
     4141 gtcagcgtca tcctcgtcct ctggatcggg tacaaatggt tcaagtgcat caggtaatgc
     4201 ccagcaaaag gaggcctggc caagcttatc ggtcccgccc atcaatggca aagaggcggc
     4261 tgccactgcg accagttcca gtgggaaaag taaacgggag aagctgcgca acgagaagag
     4321 gcacgagaag aacaaggcga agaataagaa cggcagcaat acaaacgcca atgcctcaaa
     4381 caaggagaac tatgttcccg aaacgagaag cagcacaagt actgagacat tcgcagaggc
     4441 tacggcggat gcaccggctt ccaccaaggc agaaccgccg caagcctcta gcaatagaac
     4501 gagggcggat cgtggaaaag atcggaccac ggctagtgca aaggagcaga agaagggcaa
     4561 ggaaactgct ccagcacctg cagcaagtaa accggcggag cgggttgaaa caagcgagag
     4621 tacaataaga caaaagaagg cggaagtaac cgaaagctgt gaagataact taccacaaaa
     4681 gagattagcg ggaacaaacg ttcaaagatc tgtgagctct tgtagcgaaa atagcgaagg
     4741 acacgtctct gagagtagct taagtgagaa gagtttaact ggtgattatg tggaggaaaa
     4801 gtgcaatagt gtgaattcgg aaagccagca agaaagtgta aagtttcagg aggagctcga
     4861 aaagagcaac gaggcaattg tcgaggccga aacgattctg ccaacagctg agtcctctga
     4921 ggacatcagt cccgctgcag tggcgccagg caatggagaa gacgaaggat gcttacccgt
     4981 tgttgatcca gtcgaaccac cgagtctggc ggataatgga agcagagtaa cagatgcgct
     5041 atctaagctg aacattttcg atgacacgcc tagctttttc acatcgccat cattccagca
     5101 agcacccata ttaaagaata aattggacct ggagatgcgc caatcacacc ttccggatct
     5161 ggtgaatgat atcgacggca ttcagaaagc gagcaacacg aatgaatggg aagaggcttt
     5221 caagaacgtc atgatgggaa atacccaaca tatggaagaa cagttgctgc aacaacagca
     5281 cctgcaacaa caccaaaacc tgcgtcatca attggtcctg cagcaagaag aatttttgcg
     5341 tatgcaggag cttcaaaagc gtaataattt cgctacccag attaatggac cggctaacga
     5401 ctttttgcga gcctacgaat tgcgcgccca ggctaacgcg attatccaac agcagctact
     5461 acagcaacat gcgggagaga acttgttcgg aggaaatatg tccaaatttt ttgactttca
     5521 caagtcgcaa ccccaatcgc atcaccaata ccttaacggc caccccccgc agattaacgg
     5581 taacggagcc gtccccgaac cccagcgagt cgctgcttcg ctggagtcca accgcttgaa
     5641 cagcccgttc gtggagaacg gcttgatcaa tagccaacaa caacaacagc aacagcagaa
     5701 acaacggatg atgggcatgt acgagttcat gccaccgaat acccagagtc aacaaaaccg
     5761 cttcacgcaa aatagcattg tggatgacga tctaggcttt gatcccttcg ttgagaccca
     5821 aaagggactt gctgaactta tggaaaatga ggtggtccaa aaacagagta ttaataatga
     5881 gaaccccttg ccgaagttgc cgccacagcc tcaagttcca ccccatccgc aattggtgga
     5941 caacctgcag cgagctcgca tgccgccgcc aggcttcaat cacgtcaaca cattgggctt
     6001 gggcggtgca tccagactgc agctcaccag caaaataatg cccttcatga acatgcccgt
     6061 taatggggtg ggcaacagcg gtgcccaggg acagcaccaa atgccaatgg gtgtcaactg
     6121 gaatgcgccg atgggcatgc accagaaccc gggacagccc gtgggtgatt cgcaattgca
     6181 gcatccaatg gctcacaaca aggtttacaa caacagtgat tggacttcaa tggatccggc
     6241 tatactttcg tttagacagt attcttcttt tccacaaaac caaattcccc cacatcctca
     6301 gcagcaacag gatttatttt tgcagcattt ggctcaacag caaaattccc agagtggtgg
     6361 tttcaacaac caggcacagc aaatgcttcc tatggggatg cccaatagtt tgctgaacgg
     6421 acaacaaacg cagccgccac aggtcaatgc caatgttcag ggcatgcttg agtttttaaa
     6481 aagccgtcaa ttcgtttagt tcgaaggtaa gcctatccct aaccctctcc tcggtctcga
     6541 ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc
     6601 gactgtgcct tctaaggcct gagctcgctg atcagcctcg atcgaggatc cagacatgat
     6661 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     6721 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     6781 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     6841 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt cgaccgatgc
     6901 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     6961 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     7021 gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg
     7081 cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac
     7141 gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct
     7201 tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg
     7261 gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc
     7321 agggacagct tcaaggatcg ctcgcggctc ttaccagcca gcaaaaggcc aggaaccgta
     7381 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     7441 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     7501 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7561 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     7621 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7681 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7741 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7801 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     7861 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7921 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7981 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     8041 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     8101 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     8161 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     8221 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     8281 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     8341 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     8401 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     8461 acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     8521 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     8581 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8641 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     8701 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8761 gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8821 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8881 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8941 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     9001 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     9061 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     9121 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     9181 cattaaccta taaaaatagg cgtatcacga ggccctttcg t
//
