LOCUS       sequence-288736-        9596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmSmg5-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9596)
  AUTHORS   Huntzinger E, Kashima I, Fauser M, Sauliere J, Izaurralde E
  TITLE     SMG6 is the catalytic endonuclease that cleaves mRNAs containing 
            nonsense codons in metazoan.
  JOURNAL   RNA. 2008 Dec . 14(12):2609-17.
  PUBMED    18974281
REFERENCE   2  (bases 1 to 9596)
  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: "RNA. 2008 
            Dec . 14(12):2609-17."
COMMENT     Sequence Label: pAc5.1B-EGFP-DmSmg5-siRNAres-V5His6_F
FEATURES             Location/Qualifiers
     source          1..9596
                     /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             6881..6922
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6932..6949
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6972..6989)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7022..7156
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7046..7065)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7100..7119
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7778..8366)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7858..7877)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8537..9328,9329..9397))
                     /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     9160..9179
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9398..9502)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9570..9588
                     /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 ttgaattcta tggaggtgac attcagctcg ggcggatcat cgaacgcctc
     3361 cagcgagtgc gccatagacg gcggcacgaa tcgctgtcgc ggcttggagc ccaacaatgg
     3421 aacctgcatc ctcagccagg aggtaaagga tctttacaga tccctctaca ccgcctccaa
     3481 gcaactggac gatgccaaga ggaatgtgca gtcggtggga cagctgttcc aacatgagat
     3541 tgaggaaaag cgctcgctcc tagtgcagct gtgcaagcag atcatcttca aggactacca
     3601 gtcggtgggc aagaaggtgc gcgaggtgat gtggcgacgt ggatactacg agttcattgc
     3661 cttcgtgaag aagaactggc acaagcagtg ccaggatgcg gccacggccc aagagaacat
     3721 gcaaaagctg gagcgattcc tctgcgctgg catcttcaac tacaaacgat tgtcggcacg
     3781 catggaagag atctatgatc tggatctaaa gtatttgatt gatttctcca taatcagtga
     3841 tggctacata agcgacatga tcggagacac catggagtca tcgcactcgg gcacccacgc
     3901 ggtggacaag agtctggagg ccgtgtcctt tgcactcgac accatacact catcgctgct
     3961 cagtctgggc gatcttcatc gctactttct agactttcgc atcgacagca aactaagcat
     4021 aagcaaggag caggtggcca agtattatct ggaggccttc aagcttaatc cggccattgg
     4081 catggcccag aaccagttgg gtacactgca ctacggacag aatcatgacc tggactccac
     4141 ttaccactac ttgtactccc ttgtgtgcat cataccattc gaactaagcg aaaataatct
     4201 taacaagctg ttcgccaagc acaccgaata cctagagcgt atggatccag aaaaactaga
     4261 cttcaacatc gaggacttct ttgcccgttt ctatctgatt gtggacatat ttttcttcga
     4321 caaggaagtg ccggacttta acgctctgtg ccactgtgtg ctgattgatc tacgcaagat
     4381 cctgtgctcc cagcagctgc gcgttccgga gccaagcatc ttcaagattg tctccattct
     4441 gttcttctgt ctgtccaagc ttaagatgat caactcacca aaggtttact cgctgcacgc
     4501 ctttctggtt gctgcctgtt ccgatctgat ggacgcctgc atcgtcagca tcgagcagac
     4561 aatcctggct cgcgcccaag acaacgagcg cttccaggcg gagtacgagg aacgctttca
     4621 ggagttcgat actgtggtgc gcaagtcccg gaatgagtac aagaactggc tagctgcagt
     4681 aggcgaatgt gagcgcaagg acaagaagtt gatgccacgc ccgcactctc taaaggattg
     4741 cagttcggat tcgcgggaca gccagcactc tggcgaggag gcaaagaccc aggaggctgc
     4801 tgacggcgac aagatcggtg aggccgtttc cacgcgctcc agtgacgagg ccaaggagtc
     4861 cgaccttaag accccgctct cctgcaagag caagcgacgg ggaatgcgtc tacgccgccg
     4921 acgccgcaag atcgcccagt cggacgacag ctcctgctat gacagcgaaa gcgaactgga
     4981 cacagacttt tacagcgacg aggaggatta cacagatcta agttctaact ttgacaccga
     5041 tttcagcgac attgatgcag ctgatcctgg ggagccatgc ggcgaagagc gctatgcagc
     5101 agctaattac agcaaaaagg agcgcaaagc tggcggcggc ggcggaggag gagtcggtgg
     5161 cgtggtgtac tccgacaacg aagacgttgt tattgaggag gagaaaatcg tgtatcccaa
     5221 cgaagtagag cgcagatcca actcacagga ggcggactct gaggagttgc tgcgcagctt
     5281 tgaggtactg cagctgaact ttaagagcgc agaggatgct caaagcaagc cggtggcacc
     5341 tccgcccgtc agcttttcag agccaccaaa gaagctggtt tatcgcaaca agtacaccaa
     5401 gatcaatccc aacatagtca ttgactgtct gcacaatgaa ccgaccatta gagccctgaa
     5461 aatgcttttt gactggctgc gaatcaaccc agagattctc atcggctgct accactcgaa
     5521 tccggagttt gtgcacaaga tcatgaagct tctcaactac tgcaacatcg acatcttcac
     5581 caggaaggtt tacttcgagc gcgaccttct gaccactgcc aacgtgcgtg attctctggt
     5641 cgagttattc gatgtgcgcg ccaacgttcc gctcaccgag gactttgctt tcaagaactt
     5701 tgacatcttc cagagcaccc agattacact ggactgggag accattatcc gggagcgtgt
     5761 cactccccag gaagagtcca ttctccgtat tttcaaaatg gtcgactttg gctttttcat
     5821 ttgcaagcag aagaagttta actacagatt ctgcgtcaag acgcgtcgtt tcaccgagac
     5881 ccaaaagaag aaacagcgcg aggagaagaa gggaggcgga tcacgtcgcc gcgagcgtcg
     5941 taccgctccg aagaccaacc gaacaaagcg gaacgaagga cgcacacgtc gctactccga
     6001 tcgcaagaac ggcatcgtcc gcaagagcta caccagtaac gaggaaaagg acaccgtgga
     6061 ggagtgccgc aagatgccac gcaagggtta cctaaggaat cgcaatgccg agaaggcggc
     6121 tgctgttata accagtgcca cgggaacctc tgtcgccggc ggaacctccg cctccacaac
     6181 cacaaacagt gtcatcgaga aactgaattt gcagtccagc tcgggggccg acgaagagcg
     6241 ctcggaggcc aagtcgaaga agtgtgagct catgggcaag ctgtggctgc aaaacgaggt
     6301 ggagactttg gaggaggagc tgcgggacag tcccgctcat gtggtgatgt ccccatttgt
     6361 ggtggtcgat tcgaaggcat tgaccgaata caatggcata gtcaagtcgc tggtgaggac
     6421 caagaagtgc attgtcctca ttccaaatgc agtgctcaat gagttggatg acttgaagaa
     6481 acgcagcgag aatgtgcgcc atgtcatccg ttggctggag caggagttta aacagggcaa
     6541 taggcacttg cgtgcccagc gcgataacga aagtcagccg ctatccctgc tgaagatatc
     6601 acgcaagatg gatcgtgacg cctcagtttt ccttcaaatt gcccagttct gcaatcactt
     6661 ggtggccaac catgccgatc cgcacgtcag cgaactgcag aagagcgtcg tcacgtactt
     6721 gtccggcgat aatttgcacg agaagaaccg ccagcaacag aacttcagct acaccggcat
     6781 ccttgactcg atacccgtac gctatgccca aatccagagc ttctacgcaa aatttaaggc
     6841 caacaaaaaa cgctcgagtc tagagggccc gcggttcgaa ggtaagccta tccctaaccc
     6901 tctcctcggt ctcgattcta cgcgtaccgg tcatcatcac catcaccatt gagtttaaac
     6961 ccgctgatca gcctcgactg tgccttctaa ggcctgagct cgctgatcag cctcgatcga
     7021 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     7081 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     7141 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     7201 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
     7261 tcagtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
     7321 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
     7381 tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga
     7441 tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg
     7501 gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg
     7561 cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga
     7621 ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg
     7681 tagatgacga ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agccagcaaa
     7741 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7801 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7861 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7921 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
     7981 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     8041 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     8101 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     8161 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     8221 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     8281 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     8341 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     8401 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     8461 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     8521 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     8581 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     8641 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     8701 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     8761 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     8821 ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt
     8881 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     8941 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     9001 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     9061 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     9121 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
     9181 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     9241 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     9301 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     9361 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     9421 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     9481 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     9541 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgt
//
