LOCUS       sequence-288675-        8918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmSMG6-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8918)
  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 8918)
  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-DmSMG6-siRNAres-V5His6_E
FEATURES             Location/Qualifiers
     source          1..8918
                     /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             6203..6244
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6254..6271
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6294..6311)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6344..6478
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6368..6387)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6422..6441
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7100..7688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7180..7199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7859..8650,8651..8719))
                     /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     8482..8501
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8720..8824)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8892..8910
                     /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 tgagtcgaaa gggaacacga gaacaaaatg atatatcgct
     3361 gataattggg acgctgccga aacgcctgca gaacaaaatc gcggccagct gcagccacgc
     3421 agcagcagcg gcaacaacaa caaaggcgtt ggagacggaa aacgctgaga atgcggagcc
     3481 cgtggctacg acagcaaatg cttcaaggcc agtcgaaatc ctgagcaatg ctagcacagg
     3541 aggaggcaaa cccgatgccc agtcaaagca acccgttttg gataccagcg ggaaaatggc
     3601 gtgggagggc gggattccag gcatcctgcg cgtgaccacc tcggtggcgg gcgatgttaa
     3661 caggaacgga cagggtcagg gcaagaacaa gagcggtaac agtgctcggc gcaggggtcg
     3721 caaggacaag gagcacgcta cgcagtcgcc agtgacgcgc acgcgcttga ccaccgagca
     3781 tccggattgg tttgagccgc tgttgggtct gccccatgcc aagaatatag tccatctgta
     3841 cgatcagctc atggtcctca atcagaggaa tctcatactg ataaactgga agaacttctg
     3901 ctacattcac gaccagctgc aggacgcttt caagcaactt cttttggagc aacttaagtt
     3961 cgtgtgcgag cacaaggttg atgtcttctt ttggaaacta ctcttctaca atgtgcggaa
     4021 ttacttgaag cgccagcaaa cggaccaggc gcacacacat acgctccttc tcatcgagca
     4081 agctatcaag ttttaccgca tgctctacga caagctgatg gccaagtgcg tggcatcgtc
     4141 gcgctgtgac agcgcactta aagtggtggc ccagaggctg ctcatctgtt tgggcgatct
     4201 gacgcgctac cgggttaacc atgtcaaggc cacagattac atggaggcgg ccaggtacta
     4261 tcagagggcg caggaactgg tacctggcaa cggggcacct ttcaaccagc tggcggtcat
     4321 ttccatttat caccacaagc gcttcgatgc agtttattac tatgtccgca gcttgttgac
     4381 ttcgaactct atccaatctg ccaaggaaag cctgctggat ttgttcgacg agattcggcg
     4441 taagtacgag gagaccgaga tgaagcaatc tcccatgcac tatgcagctg gatcgaagaa
     4501 tcaaaagtcg aagcagatgc gcaaggaggt atggatctat ccggatggga tcagacgcct
     4561 acatcgcacc gatgacaagg gcaacaaggc gaagggcaac aaggccacta tggcagaggt
     4621 caatcgttac gaggagatgc cgccggagga gattcttcct cgcattgttt ccctttatct
     4681 gtacttatta ggcaaactct acaccggcac agatgtggac tcgttatacc cgttgctaag
     4741 gaaactgcaa atccaaatag gcgtcgcttt aaggcacgaa aacctgttgt cacgctccaa
     4801 actgctgaaa atagtagcac tcaacttgtt cgtcgtcgag cacaacaaac ccaaaacgtc
     4861 acgccgcgag atgcgttatc acagcttcaa ttttgccaac tcattctttg gactcatgat
     4921 gaagaaaacg aaccagctgc tggctggttt tgtagaggat tctacaaatg tccagtgtct
     4981 cccagaagaa gactttgcga ctgtgaacac atacctacag ttcgtgaatg tttatgtacg
     5041 ttggttgagc atcagtttgg acgtgtggga acccgtcaga tcggaagaac actctttcat
     5101 cgattgctgg gctgagctaa caatcctttt tgagtacatt gaatgcttaa tggggaagca
     5161 caaacttgag aaaaacgaaa tactgttgga tgaggatgtg gcccttagag gtttcactcc
     5221 gttgggtcgt gaaacgctga gcatggacta tctcaaaaga ggatccgaac gtcttcagtt
     5281 tttcgaacgt gttcgtagga ttggtcagtt ccaggaatac tacgtccagc atcaagaggc
     5341 tctccagcag ccgttggaat caagcttcac tgtcgagcac ctaaacgcag cgatgaagaa
     5401 cgcattggaa aactatgatg ctgaaagttg tggtcttgcg gcgaacgttt caaagattca
     5461 atgtaaagcc gttgaggaat tgttgtgtcg ttcaactgat tccgatgttt ccgagctgtg
     5521 caagctgaag aaagagcttg aggccaaggc tagggttaag cagatatgca ccagcaagtt
     5581 ggaggacatt ctcaagttcg tggacacgaa aatatacatc gaggtgcgac cgcgctatct
     5641 tctaccggat accaactgct ttatcgactg ccttgaagac tttgaaaaac tgtccacgga
     5701 gtttaaacgc tacacactta ttataccact gacagtggtt aaggaactgg atggcctctc
     5761 caagggcgtt aagttggact cgtaccgcag ttcgaagcag acgcagcgca tccaccactt
     5821 tgacgaggtg tcgtcgcggg ccaaaaagtc gttagagttc atcaagtcgg ccaaaggcaa
     5881 tgtgaaatgt gctaccacaa agggctcgtt tgtcaatgca tcggtgtttg ccctagttga
     5941 ggaggaatat ttgtccaatg acgacaagat tctggcaacg gcaatggctg tctcgaaaac
     6001 ggcaaaaacc gaacaatgca cagcagatgg caagtgtttc attcaaacgg aattggtgct
     6061 cattacaacc gatcgtaatc tgcgtgtgaa ggctctttct cgcaatttgg ccgtcagcgc
     6121 gttggatgag tttctgcaat gggccaagga ttgccgcgaa tcaaccggcg gccgctcgag
     6181 tctagagggc ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc
     6241 tacgcgtacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     6301 tgtgccttct aaggcctgag ctcgctgatc agcctcgatc gaggatccag acatgataag
     6361 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     6421 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     6481 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     6541 aagcaagtaa aacctctaca aatgtggtat ggctgattat gatcagtcga ccgatgccct
     6601 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg
     6661 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg
     6721 tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg
     6781 tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt
     6841 tcggcgagaa gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc
     6901 tggcgttcgc gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg
     6961 gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg
     7021 gacagcttca aggatcgctc gcggctctta ccagccagca aaaggccagg aaccgtaaaa
     7081 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7141 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7201 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7261 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     7321 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7381 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7441 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7501 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     7561 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7621 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7681 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7741 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7801 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7861 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7921 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7981 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8041 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8101 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8161 ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8221 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8281 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8341 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8401 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8461 cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8521 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8581 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8641 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8701 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8761 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8821 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     8881 taacctataa aaataggcgt atcacgaggc cctttcgt
//
