LOCUS       sequence-355929-        8577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmNot3-F818A-dsRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8577)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 8577)
  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"
COMMENT     Sequence Label: pAc5.1B-EGFP-DmNot3-F818A-dsRNAres_Q
FEATURES             Location/Qualifiers
     source          1..8577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2566
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2586..2588,2589..2591,2592..3302)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2586..2588,2589..2591,2592..3302)
                     /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(2631..2652)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2892..2911)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3239..3260
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5883..5924
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5934..5951
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5974..5991)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6024..6158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6048..6067)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6102..6121
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6759..7347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6839..6858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7518..8309,8310..8378))
                     /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     8141..8160
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8379..8483)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8551..8569
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcaagaat taattctata ttctaaaaac acaaatgata cttctaaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat aatgaatatg tacacattcg
      241 agaaagaagc gatcaaagaa gcgtcttcgg gcggagtagg agaatgcgga ggagaaggag
      301 aacgagctga tctagtatct ctccacaatc caatgccaac tgaccaactg gccatattcg
      361 gagcaatttg aagccaattt ccatcgcctg gcgatcgctc cattcttggc tatatgtttt
      421 tcaccgttac ccggggccat tttcaaagac tcgtcggcaa gataagattg tgtcactcgc
      481 tgtctctctt catttgtcga agaatgctga ggaatttcgc gatgacgtcg gcgagtattt
      541 tgaagaatga gaataatttg tatttatacg aaaatcagtt agtggaattt tctacaaaaa
      601 catgttatct atagataatt ttgttgcaaa atatgttgac tatgacaaag attgtatgta
      661 tataccttta atgtattctc attttcttat gtatttataa tggcaatgat gatactgatg
      721 atattttaag atgatgccag accaaaaggc ttgaatttct gcgtcttttg ccgaacgcag
      781 tgcatgtgca attgttgttt tttggaatat tcaattttcg gactgtccgc tttgatttca
      841 gtttcttggc ttattcaaaa agcaaagtaa agccaaaaaa gcgagatggc aataccaaat
      901 gcggcaaaac ggtagtggaa ggaaaggggt gcggggcagc ggaaggaagg gtggggcggg
      961 gcgtggcggg gtctgtggct gggcgcgacg tcaccgacgt tggagccact cctttgacca
     1021 tgtgtgcgtg tgtgtattat tcgtgtctcg ccactcgccg gttgtttttt tctttttatg
     1081 ctgcgctctc tctagcgcca tctcgcttac gcatgctcaa cgcaccgcat gttgccgttt
     1141 ccttttatgc gtcattttgg ctcgaaatag gcaattattt aaacaaagat tagtcaacga
     1201 aaacgctaaa ataaataagt ctacaatatg gttacttatt gccatgtgtg tgcagccaac
     1261 gatagcaaca aaagcaacaa cacaggtggc tttccctctt tcactttttg tttgcaagcc
     1321 gcgtgcgagc aagacggcac gaccggcaaa cgcaattacg ctgacaaaga gcagacgaag
     1381 ttttggcgaa aaacatcaag gcgcctgata cgaatgcatt tgcaataaca attgcgatat
     1441 ttaatattgt ttatgaagct gtttgacttc aaaacacaca aaaaaaaaaa ataaaacaaa
     1501 ttatttgaaa gagaattagg aatcggacgc ttatcgttag ggtaacaaca agaaatgctt
     1561 actgagtcac agcctctgga aaactgccgc aagccagaga gagagagaaa aagagggaga
     1621 gcagcttaga ccgcatgtgc ttgtgtgtga ggcgtctctc tcttcgtctc tgttgcgcaa
     1681 acgcatagac tgcactgaaa aaatcgatta cctatttttt atgaatgaat atttgcacta
     1741 ttactattca aaactattaa gatagcaatc acattcaata gccaaatact ataccacctg
     1801 agcgatgcaa cgaaatgatc aatttgagca aaaatgctgc atatttagga cggcatcatt
     1861 atagaaatgc ttcttgctgt gtacttttct ctcgtctggc agctgtttcg ccgttattgt
     1921 taaaaccggc ttaagttagg tgtgttttct acgactagtg aatgccctac tagaagatgt
     1981 gtgttgcaca aaatgtccct ggaataacca atttgaagtg cagatagcag taaacgtaag
     2041 ctaatatgaa tattatttaa ctgtaatgtt ttaatatcgc tggacattac taataaaccc
     2101 actataaaca catgtacata tgtatgtttt ggcatacaat gagtagttgg ggaaaaaatg
     2161 tgtaaaagca ccgtgaccat cacagcataa agataaccag ctgaagtatc gaatatgagt
     2221 aacccccaaa ttgaatcaca tgccgcaact gataggaccc atggaagtac actcttcatg
     2281 gcgatataca agacacacac aagcacgaac acccagttgc ggaggaaatt ctccgtaaat
     2341 gaaaacccaa tcggcgaaca attcataccc atatatggta aaagttttga acgcgacttg
     2401 agagcggaga gcattgcggc tgataaggtt ttagcgctaa gcgggcttta taaaacgggc
     2461 tgcgggacca gttttcatat cactaccgtt tgagttcttg tgctgtgtgg atactcctcc
     2521 cgacacaaag ccgctccatc agccagcagt cgtctaatcc agagaccccg gatcggggta
     2581 ccatcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2641 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     2701 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     2761 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     2821 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     2881 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     2941 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     3001 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     3061 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     3121 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3181 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     3241 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3301 agccaagctt gatggctgcg acgagaaaat tgcaaggcga aattgatcga tgtttaaaaa
     3361 aggtagcaga gggcgttgaa acgtttgagg atatttggaa aaaagtacac aatgcgacaa
     3421 acaccaatca gaagcaaaaa catctgcagg agaagtatga ggcagacctc aaaaaggaaa
     3481 tcaaggaatt gcaacgactt cgcgaccaaa ttaaatcatg gattgcatca gctgaaatta
     3541 aggataagag ctcccttctt gagaatcgac gtctcattga aacacaaatg gaacgcttta
     3601 aggtggttga gcgcgagacg aaaacaaaag cctactccaa ggaaggcttg ggggctgcac
     3661 aaaaaatgga cccagctcaa cgcatcaagg atgatgcgcg taactggcta acgtccagta
     3721 tttcgagctt gcagattcag atcgatcaat acgaatccga aattgagagc ctgctggccg
     3781 gaaagaaaaa gcgcctagac cgtgataaac aagaacgtat ggacgatctt cgcggcaaac
     3841 tggatcgtca caaattccat attaccaaat tggagacatt gctgcgattg ttggataacg
     3901 atggagtgga ggctgaacaa gttaataaaa ttaaggatga cgtggagtat tatatcgaca
     3961 gtagccagga acccgatttc gaagagaacg aatttattta tgacgacatt atcggattgg
     4021 acgaggtcga actttcggga accgccacga ccgattccaa taattccaat gaaacctccg
     4081 gaagtccctc ctcggtgacg agcggcggat cgcccagtca aagcccagtg accgtccagc
     4141 agatcctaaa tacaagcagc caaggtgctg cctcgtccgg ctcctccgct gcctccgctg
     4201 cgttgttcca acaacaattg accgccgctc agtccaacgg aaataacgtc ggatatgcct
     4261 ccgatacgtc ggctgcctca tcctcagcca ccacctcaac cgacccagcc ggaggaacgg
     4321 tcgccgttaa ctgcgtcggc ggtcttgctg acaagcgtaa taagagcagc gaaagcaatg
     4381 ccctaaaatt gaaaccacaa cctcatcaac ttataaaacc tacccctgta cgagccacgg
     4441 ccaagctgcc tctgagcagt gatacccaag tcaataagat tgtcagttca actcccagca
     4501 aaaatcagca acagctgcct acagcagctt caatcgttgc aacctccgct atgcaatcgc
     4561 aaagcagtat cggaagttgc agctccactg gaggaacggg agcatctcaa tcagcttcta
     4621 gtggtaacaa tcctggtaac aatcctgcgg ttcagccaaa tgctccaacg cctggtcaaa
     4681 gcggcattgc cgctgcagca gcttcgacta atgttgtttc ggcgacgatt gtgagcagcg
     4741 ccaatgttca aggccaatct gtgatccaac caacaccaac aattgcattt gctgcagtag
     4801 caaaacacaa tacatcactc ctggaaaatg gtcctgtttt gcaacagcaa ctagcagtaa
     4861 ctcctactgt ggcagctatc gtaggagctg gaacgcaggc gcagcaaaaa catgtgccac
     4921 cattatccaa tcttcaaaca aattcccctc acatacaaaa tggtcttcct gtctcggaca
     4981 gtactaatga caacagctgc aatgtcgtcg atacgatttc tctaaaaaca atggcacagg
     5041 atgctattaa tcgatctgcg atcgatccta attcactgaa tcagcagcaa acaagcagca
     5101 tcgatctgag acagccgcag tcgcagaaat cacttctcca acatttcaac tctgaaacca
     5161 acacgaatca acagcaattg acatcgcagc agcagcaaca gcttcagaat aactcccttg
     5221 ctgcgaccac aggttctaac aacggaactt caactggtag tggtctcatg aatgtagcaa
     5281 acgctactgg tcagcctatc agcggcaatg ccaaaacgca tgcgtgccaa ccccaacaga
     5341 tggccacgac ggaagcgcac attcccacac tgcttggtgt gacaccactt gggccaacac
     5401 cacttcaaaa ggagcaccaa atgcagttcc aaatgatgga agctgcctac tatcacctgc
     5461 cacagcctat ggacacggaa aagctacaaa cgtactttca tcgcgcacca gtgctcacgc
     5521 catcgcacta tccccaggca cagatgccaa tatatgatac agttgagttt tatcaacgac
     5581 tctcaacgga gacccttttt tttgtgttct actatatgga gggctccaag gcccagtatc
     5641 tagcggccaa ggccttaaaa aagcagagct ggcgtttcca cacaaagtat atgatgtggt
     5701 ttcaaaggca cgaggagccg aagattatca acgatgatta cgaacagggt acgtacatct
     5761 atgctgatta tgaaaagtgg agtcagcgaa agaaggaggg attcaccttt gaatacaagt
     5821 acttagagga caaggagctg aattgaggcg gccgctcgag tctagagggc ccgcggttcg
     5881 aaggtaagcc tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc
     5941 accatcacca ttgagtttaa acccgctgat cagcctcgac tgtgccttct aaggcctgag
     6001 ctcgctgatc agcctcgatc gaggatccag acatgataag atacattgat gagtttggac
     6061 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     6121 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     6181 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     6241 aatgtggtat ggctgattat gatcagtcga ccgatgccct tgagagcctt caacccagtc
     6301 agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt
     6361 atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg cgaggaccgc
     6421 tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc
     6481 ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt
     6541 atcgccggca tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc
     6601 tggatggcct tccccattat gattcttctc gcttccggcg gcatcgggat gcccgcgttg
     6661 caggccatgc tgtccaggca ggtagatgac gaccatcagg gacagcttca aggccagcaa
     6721 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6781 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6841 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6901 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6961 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7021 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7081 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     7141 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     7201 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7261 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7321 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7381 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7441 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7501 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7561 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7621 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7681 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7741 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7801 gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg
     7861 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7921 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7981 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8041 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     8101 atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc
     8161 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     8221 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     8281 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     8341 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     8401 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8461 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     8521 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgt
//
