LOCUS       sequence-290153-        8600 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmNot3-F764E-dsRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8600)
  AUTHORS   Boland A, Chen Y, Raisch T, Jonas S, Kuzuoglu-Ozturk D, Wohlbold L, 
            Weichenrieder O, Izaurralde E
  TITLE     Structure and assembly of the NOT module of the human CCR4-NOT 
            complex.
  JOURNAL   Nat Struct Mol Biol. 2013 Nov;20(11):1289-97. doi: 
            10.1038/nsmb.2681. Epub 2013 Oct 13.
  PUBMED    24121232
REFERENCE   2  (bases 1 to 8600)
  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: "10.1038/nsmb.2681";
            journalName: "Nat Struct Mol Biol"; date: "2013-11"; volume: "20"; 
            issue: "11"; pages: "1289-97"
COMMENT     Sequence Label: pAc5.1B-EGFP-DmNot3-F764E-dsRNAres_U
FEATURES             Location/Qualifiers
     source          1..8600
                     /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             5885..5926
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5936..5953
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5976..5993)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6026..6160
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6050..6069)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6104..6123
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6782..7370)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6862..6881)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7541..8332,8333..8401))
                     /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     8164..8183
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8402..8506)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8574..8592
                     /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 ttgatggctg cgacgagaaa attgcaaggc gaaattgatc gatgtttaaa
     3361 aaaggtagca gagggcgttg aaacgtttga ggatatttgg aaaaaagtac acaatgcgac
     3421 aaacaccaat cagaagcaaa aacatctgca ggagaagtat gaggcagacc tcaaaaagga
     3481 aatcaaggaa ttgcaacgac ttcgcgacca aattaaatca tggattgcat cagctgaaat
     3541 taaggataag agctcccttc ttgagaatcg acgtctcatt gaaacacaaa tggaacgctt
     3601 taaggtggtt gagcgcgaga cgaaaacaaa agcctactcc aaggaaggct tgggggctgc
     3661 acaaaaaatg gacccagctc aacgcatcaa ggatgatgcg cgtaactggc taacgtccag
     3721 tatttcgagc ttgcagattc agatcgatca atacgaatcc gaaattgaga gcctgctggc
     3781 cggaaagaaa aagcgcctag accgtgataa acaagaacgt atggacgatc ttcgcggcaa
     3841 actggatcgt cacaaattcc atattaccaa attggagaca ttgctgcgat tgttggataa
     3901 cgatggagtg gaggctgaac aagttaataa aattaaggat gacgtggagt attatatcga
     3961 cagtagccag gaacccgatt tcgaagagaa cgaatttatt tatgacgaca ttatcggatt
     4021 ggacgaggtc gaactttcgg gaaccgccac gaccgattcc aataattcca atgaaacctc
     4081 cggaagtccc tcctcggtga cgagcggcgg atcgcccagt caaagcccag tgaccgtcca
     4141 gcagatccta aatacaagca gccaaggtgc tgcctcgtcc ggctcctccg ctgcctccgc
     4201 tgcgttgttc caacaacaat tgaccgccgc tcagtccaac ggaaataacg tcggatatgc
     4261 ctccgatacg tcggctgcct catcctcagc caccacctca accgacccag ccggaggaac
     4321 ggtcgccgtt aactgcgtcg gcggtcttgc tgacaagcgt aataagagca gcgaaagcaa
     4381 tgccctaaaa ttgaaaccac aacctcatca acttataaaa cctacccctg tacgagccac
     4441 ggccaagctg cctctgagca gtgataccca agtcaataag attgtcagtt caactcccag
     4501 caaaaatcag caacagctgc ctacagcagc ttcaatcgtt gcaacctccg ctatgcaatc
     4561 gcaaagcagt atcggaagtt gcagctccac tggaggaacg ggagcatctc aatcagcttc
     4621 tagtggtaac aatcctggta acaatcctgc ggttcagcca aatgctccaa cgcctggtca
     4681 aagcggcatt gccgctgcag cagcttcgac taatgttgtt tcggcgacga ttgtgagcag
     4741 cgccaatgtt caaggccaat ctgtgatcca accaacacca acaattgcat ttgctgcagt
     4801 agcaaaacac aatacatcac tcctggaaaa tggtcctgtt ttgcaacagc aactagcagt
     4861 aactcctact gtggcagcta tcgtaggagc tggaacgcag gcgcagcaaa aacatgtgcc
     4921 accattatcc aatcttcaaa caaattcccc tcacatacaa aatggtcttc ctgtctcgga
     4981 cagtactaat gacaacagct gcaatgtcgt cgatacgatt tctctaaaaa caatggcaca
     5041 ggatgctatt aatcgatctg cgatcgatcc taattcactg aatcagcagc aaacaagcag
     5101 catcgatctg agacagccgc agtcgcagaa atcacttctc caacatttca actctgaaac
     5161 caacacgaat caacagcaat tgacatcgca gcagcagcaa cagcttcaga ataactccct
     5221 tgctgcgacc acaggttcta acaacggaac ttcaactggt agtggtctca tgaatgtagc
     5281 aaacgctact ggtcagccta tcagcggcaa tgccaaaacg catgcgtgcc aaccccaaca
     5341 gatggccacg acggaagcgc acattcccac actgcttggt gtgacaccac ttgggccaac
     5401 accacttcaa aaggagcacc aaatgcagtt ccaaatgatg gaagctgcct actatcacct
     5461 gccacagcct atggacacgg aaaagctaca aacgtacttt catcgcgcac cagtgctcac
     5521 gccatcgcac tatccccagg cacagatgcc aatatatgat acagttgagt tttatcaacg
     5581 actctcaacg gagacccttt ttgaggtgtt ctactatatg gagggctcca aggcccagta
     5641 tctagcggcc aaggccttaa aaaagcagag ctggcgtttc cacacaaagt atatgatgtg
     5701 gtttcaaagg cacgaggagc cgaagattat caacgatgat tacgaacagg gtacgtacat
     5761 ctattttgat tatgaaaagt ggagtcagcg aaagaaggag ggattcacct ttgaatacaa
     5821 gtacttagag gacaaggagc tgaattgagg cggccgctcg agtctagagg gcccgcggtt
     5881 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca
     5941 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctaaggcctg
     6001 agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg atgagtttgg
     6061 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     6121 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     6181 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     6241 caaatgtggt atggctgatt atgatcagtc gaccgatgcc cttgagagcc ttcaacccag
     6301 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     6361 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
     6421 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg
     6481 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
     6541 ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag
     6601 gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt
     6661 tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt caaggatcgc
     6721 tcgcggctct taccagccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6781 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6841 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6901 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6961 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7021 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7081 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7141 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7201 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     7261 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7321 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7381 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7441 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7501 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7561 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7621 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7681 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7741 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7801 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
     7861 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7921 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7981 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8041 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8101 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
     8161 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8221 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8281 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8341 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8401 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8461 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8521 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     8581 gtatcacgag gccctttcgt
//
