LOCUS       sequence-355422-        7259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmeIF4A-dsRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7259)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7259)
  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-DmeIF4A-dsRNAres_AA
FEATURES             Location/Qualifiers
     source          1..7259
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..2532
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2552..2554,2555..2557,2558..3268)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2552..2554,2555..2557,2558..3268)
                     /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(2597..2618)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2858..2877)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3205..3226
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4532..4573
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4583..4600
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4623..4640)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4673..4807
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4697..4716)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4751..4770
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5408..5996)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5488..5507)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6167..6958,6959..7027))
                     /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     6790..6809
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7028..7132)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7200..7218
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aatgatactt ctaaaaaaaa atcatgaatg gcatcaactc tgaatcaaat ctttgcagat
       61 gcacctactt ctcatttcca ctgtcacatc atttttccag atctcgctgc ctgttatgtg
      121 gcccacaaac caagacacgt tttatggcca ttaaagctgg ctgatcgtcg ccaaacacca
      181 aatacataat gaatatgtac acattcgaga aagaagcgat caaagaagcg tcttcgggcg
      241 gagtaggaga atgcggagga gaaggagaac gagctgatct agtatctctc cacaatccaa
      301 tgccaactga ccaactggcc atattcggag caatttgaag ccaatttcca tcgcctggcg
      361 atcgctccat tcttggctat atgtttttca ccgttacccg gggccatttt caaagactcg
      421 tcggcaagat aagattgtgt cactcgctgt ctctcttcat ttgtcgaaga atgctgagga
      481 atttcgcgat gacgtcggcg agtattttga agaatgagaa taatttgtat ttatacgaaa
      541 atcagttagt ggaattttct acaaaaacat gttatctata gataattttg ttgcaaaata
      601 tgttgactat gacaaagatt gtatgtatat acctttaatg tattctcatt ttcttatgta
      661 tttataatgg caatgatgat actgatgata ttttaagatg atgccagacc aaaaggcttg
      721 aatttctgcg tcttttgccg aacgcagtgc atgtgcaatt gttgtttttt ggaatattca
      781 attttcggac tgtccgcttt gatttcagtt tcttggctta ttcaaaaagc aaagtaaagc
      841 caaaaaagcg agatggcaat accaaatgcg gcaaaacggt agtggaagga aaggggtgcg
      901 gggcagcgga aggaagggtg gggcggggcg tggcggggtc tgtggctggg cgcgacgtca
      961 ccgacgttgg agccactcct ttgaccatgt gtgcgtgtgt gtattattcg tgtctcgcca
     1021 ctcgccggtt gtttttttct ttttatgctg cgctctctct agcgccatct cgcttacgca
     1081 tgctcaacgc accgcatgtt gccgtttcct tttatgcgtc attttggctc gaaataggca
     1141 attatttaaa caaagattag tcaacgaaaa cgctaaaata aataagtcta caatatggtt
     1201 acttattgcc atgtgtgtgc agccaacgat agcaacaaaa gcaacaacac aggtggcttt
     1261 ccctctttca ctttttgttt gcaagccgcg tgcgagcaag acggcacgac cggcaaacgc
     1321 aattacgctg acaaagagca gacgaagttt tggcgaaaaa catcaaggcg cctgatacga
     1381 atgcatttgc aataacaatt gcgatattta atattgttta tgaagctgtt tgacttcaaa
     1441 acacacaaaa aaaaaaataa aacaaattat ttgaaagaga attaggaatc ggacgcttat
     1501 cgttagggta acaacaagaa atgcttactg agtcacagcc tctggaaaac tgccgcaagc
     1561 cagagagaga gagaaaaaga gggagagcag cttagaccgc atgtgcttgt gtgtgaggcg
     1621 tctctctctt cgtctctgtt gcgcaaacgc atagactgca ctgaaaaaat cgattaccta
     1681 ttttttatga atgaatattt gcactattac tattcaaaac tattaagata gcaatcacat
     1741 tcaatagcca aatactatac cacctgagcg atgcaacgaa atgatcaatt tgagcaaaaa
     1801 tgctgcatat ttaggacggc atcattatag aaatgcttct tgctgtgtac ttttctctcg
     1861 tctggcagct gtttcgccgt tattgttaaa accggcttaa gttaggtgtg ttttctacga
     1921 ctagtgaatg ccctactaga agatgtgtgt tgcacaaaat gtccctggaa taaccaattt
     1981 gaagtgcaga tagcagtaaa cgtaagctaa tatgaatatt atttaactgt aatgttttaa
     2041 tatcgctgga cattactaat aaacccacta taaacacatg tacatatgta tgttttggca
     2101 tacaatgagt agttggggaa aaaatgtgta aaagcaccgt gaccatcaca gcataaagat
     2161 aaccagctga agtatcgaat atgagtaacc cccaaattga atcacatgcc gcaactgata
     2221 ggacccatgg aagtacactc ttcatggcga tatacaagac acacacaagc acgaacaccc
     2281 agttgcggag gaaattctcc gtaaatgaaa acccaatcgg cgaacaattc atacccatat
     2341 atggtaaaag ttttgaacgc gacttgagag cggagagcat tgcggctgat aaggttttag
     2401 cgctaagcgg gctttataaa acgggctgcg ggaccagttt tcatatcact accgtttgag
     2461 ttcttgtgct gtgtggatac tcctcccgac acaaagccgc tccatcagcc agcagtcgtc
     2521 taatccagag accccggatc ggggtaccat catggtgagc aagggcgagg agctgttcac
     2581 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2641 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     2701 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     2761 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2821 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     2881 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2941 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     3001 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     3061 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     3121 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     3181 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     3241 cactctcggc atggacgagc tgtacaagcc aagcttgaat tctatggacg atcgcaacga
     3301 aatcccgcaa gacggaccgg cttccatgga gccggaagga gtgattgaaa gcacgtggca
     3361 tgaagtctat gataattttg acgatatgaa tctgcgtgaa gaactgttgc gtggaattta
     3421 tggctacggc tttgaaaaac ccagcgctat tcaacaacgt gctattatcc cgtgcgtccg
     3481 cggccgtgac gtgatcgctc aagctcaaag cggcaccgga aaaaccgcta cgtttagcat
     3541 cgccattctg caacagattg atacctccat ccgtgaatgt caagccttga ttttggctcc
     3601 gacccgtgaa ctggctaccc aaattcaacg tgtcgtcatg gccctgggag aatatatgaa
     3661 agtccatagc catgcttgta tcggaggaac caatgtccgt gaggatgctc gtattctgga
     3721 gagcggctgt cacgtcgtcg tcggaacccc gggacgtgtg tatgatatga ttaatcgtaa
     3781 agtcttgcgt acgcaatata ttaaattgtt tgtcttggac gaagctgacg aaatgctgag
     3841 ccgtggcttt aaagaccaaa ttcaagacgt gtttaaaatg ttgccgcccg acgtccaagt
     3901 gattttgttg agcgctacga tgccccccga cgtcctggag gtctcccgct gttttatgcg
     3961 cgacccggtc agcatcctgg ttaagaagga ggaactgacc cttgagggta tcaagcagtt
     4021 ttacgtcaac gtgaagcagg agaactggaa actgggcacc ctttgcgatc tgtacgatac
     4081 gctgtccatc acccagtcgg taatcttctg caacacccgt cgcaaggtgg accaactgac
     4141 ccaggagatg tctatccaca acttcaccgt ctcggccatg cacggcgaca tggagcagcg
     4201 tgatcgcgag gtcatcatga aacaattccg ttcgggctcg tctcgtgttc tgattaccac
     4261 tgatttactg gcgcgcggta ttgatgtgca gcaggtgtcg ttggtcatca actatgatct
     4321 gccctcgaac cgcgagaact acattcatag aattggtcgc ggtggtcgtt tcggtcgcaa
     4381 gggtgttgcg atcaacttta ttacagatga tgatcgacga atccttaagg atattgaaca
     4441 gttctaccac acaacaattg aggaaatgcc tgctaatatt gccgatttga tttaaggcgg
     4501 ccgctcgagt ctagagggcc cgcggttcga aggtaagcct atccctaacc ctctcctcgg
     4561 tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc
     4621 agcctcgact gtgccttcta aggcctgagc tcgctgatca gcctcgatcg aggatccaga
     4681 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     4741 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     4801 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     4861 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcagtcgac
     4921 cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta
     4981 tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag
     5041 cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt
     5101 cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca
     5161 ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggccgac gcgctgggct
     5221 acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt ccccattatg attcttctcg
     5281 cttccggcgg catcgggatg cccgcgttgc aggccatgct gtccaggcag gtagatgacg
     5341 accatcaggg acagcttcaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5401 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5461 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5521 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5581 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5641 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5701 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5761 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5821 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     5881 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5941 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6001 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6061 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6121 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6181 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6241 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6301 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6361 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6421 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6481 ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6541 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6601 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6661 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6721 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6781 caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6841 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6901 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6961 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7021 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7081 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7141 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     7201 ataggcgtat cacgaggccc tttcgtcttc aagaattaat tctatattct aaaaacaca
//
