LOCUS       sequence-288682-        9671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmSMG6-NusA-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9671)
  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 9671)
  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-lambdaN-HA-DmSMG6-NusA-siRNAres-V5His6_E
FEATURES             Location/Qualifiers
     source          1..9671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2568
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             2588..2653
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     CDS             2678..2704
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4757..6235
                     /codon_start=1
                     /gene="nusA"
                     /product="transcription elongation protein (N utilization 
                     substance protein A)"
                     /label=NusA
                     /note="highly soluble in E. coli"
                     /translation="KEILAVVEAVSNEKALPREKIFEALESALATATKKKYEQEIDVRV
                     QIDRKSGDFDTFRRWLVVDEVTQPTKEITLEAARYEDESLNLGDYVEDQIESVTFDRIT
                     TQTAKQVIVQKVREAERAMVVDQFREHEGEIITGVVKKVNRDNISLDLGNNAEAVILRE
                     DMLPRENFRPGDRVRGVLYSVRPEARGAQLFVTRSKPEMLIELFRIEVPEIGEEVIEIK
                     AAARDPGSRAKIAVKTNDKRIDPVGACVGMRGARVQAVSTELGGERIDIVLWDDNPAQF
                     VINAMAPADVASIVVDEDKHTMDIAVEAGNLAQAIGRNGQNVRLASQLSGWELNVMTVD
                     DLQAKHQAEAHAAIDTFTKYLDIDEDFATVLVEEGFSTLEELAYVPMKELLEIEGLDEP
                     TVEALRERAKNALATIAQAQEESLGDNKPADDLLNLEGVDRDLAFKLAARGVCTLEDLA
                     EQGIDDLADIEGLTDEKAGALIMAARNICWFGDEA"
     CDS             6956..6997
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             7007..7024
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7047..7064)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7097..7231
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7121..7140)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7175..7194
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7853..8441)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7933..7952)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8612..9403,9404..9472))
                     /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     9235..9254
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9473..9577)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9645..9663
                     /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 gacgcacaaa cacgacgacg tgagcgtcgc gctgagaaac aagctcaatg
     2641 gaaagctgca aacggtggat ctcctccaca tatggcttac ccatacgatg ttccagatta
     2701 cgctcctcca agcttgaatt ctatgagtcg aaagggaaca cgagaacaaa atgatatatc
     2761 gctgataatt gggacgctgc cgaaacgcct gcagaacaaa atcgcggcca gctgcagcca
     2821 cgcagcagca gcggcaacaa caacaaaggc gttggagacg gaaaacgctg agaatgcgga
     2881 gcccgtggct acgacagcaa atgcttcaag gccagtcgaa atcctgagca atgctagcac
     2941 aggaggaggc aaacccgatg cccagtcaaa gcaacccgtt ttggatacca gcgggaaaat
     3001 ggcgtgggag ggcgggattc caggcatcct gcgcgtgacc acctcggtgg cgggcgatgt
     3061 taacaggaac ggacagggtc agggcaagaa caagagcggt aacagtgctc ggcgcagggg
     3121 tcgcaaggac aaggagcacg ctacgcagtc gccagtgacg cgcacgcgct tgaccaccga
     3181 gcatccggat tggtttgagc cgctgttggg tctgccccat gccaagaata tagtccatct
     3241 gtacgatcag ctcatggtcc tcaatcagag gaatctcata ctgataaact ggaagaactt
     3301 ctgctacatt cacgaccagc tgcaggacgc tttcaagcaa cttcttttgg agcaacttaa
     3361 gttcgtgtgc gagcacaagg ttgatgtctt cttttggaaa ctactcttct acaatgtgcg
     3421 gaattacttg aagcgccagc aaacggacca ggcgcacaca catacgctcc ttctcatcga
     3481 gcaagctatc aagttttacc gcatgctcta cgacaagctg atggccaagt gcgtggcatc
     3541 gtcgcgctgt gacagcgcac ttaaagtggt ggcccagagg ctgctcatct gtttgggcga
     3601 tctgacgcgc taccgggtta accatgtcaa ggccacagat tacatggagg cggccaggta
     3661 ctatcagagg gcgcaggaac tggtacctgg caacggggca cctttcaacc agctggcggt
     3721 catttccatt tatcaccaca agcgcttcga tgcagtttat tactatgtcc gcagcttgtt
     3781 gacttcgaac tctatccaat ctgccaagga aagcctgctg gatttgttcg acgagattcg
     3841 gcgtaagtac gaggagaccg agatgaagca atctcccatg cactatgcag ctggatcgaa
     3901 gaatcaaaag tcgaagcaga tgcgcaagga ggtatggatc tatccggatg ggatcagacg
     3961 cctacatcgc accgatgaca agggcaacaa ggcgaagggc aacaaggcca ctatggcaga
     4021 ggtcaatcgt tacgaggaga tgccgccgga ggagattctt cctcgcattg tttcccttta
     4081 tctgtactta ttaggcaaac tctacaccgg cacagatgtg gactcgttat acccgttgct
     4141 aaggaaactg caaatccaaa taggcgtcgc tttaaggcac gaaaacctgt tgtcacgctc
     4201 caaactgctg aaaatagtag cactcaactt gttcgtcgtc gagcacaaca aacccaaaac
     4261 gtcacgccgc gagatgcgtt atcacagctt caattttgcc aactcattct ttggactcat
     4321 gatgaagaaa acgaaccagc tgctggctgg ttttgtagag gattctacaa atgtccagtg
     4381 tctcccagaa gaagactttg cgactgtgaa cacataccta cagttcgtga atgtttatgt
     4441 acgttggttg agcatcagtt tggacgtgtg ggaacccgtc agatcggaag aacactcttt
     4501 catcgattgc tgggctgagc taacaatcct ttttgagtac attgaatgct taatggggaa
     4561 gcacaaactt gagaaaaacg aaatactgtt ggatgaggat gtggccctta gaggtttcac
     4621 tccgttgggt cgtgaaacgc tgagcatgga ctatctcaaa agaggatccg aacgtcttca
     4681 gtttttcgaa cgtgttcgta ggattggtca gttccaggaa tactacgtcc agcatcaaga
     4741 ggctctcggc ggccgcaaag aaattttggc tgtagttgaa gccgtatcca atgaaaaggc
     4801 gctacctcgc gagaagattt tcgaagcatt ggaaagcgcg ctggcgacag caacaaagaa
     4861 aaaatatgaa caagagatcg acgtccgcgt acagatcgat cgcaaaagcg gtgattttga
     4921 cactttccgt cgctggttag ttgttgatga agtcacccag ccgaccaagg aaatcaccct
     4981 tgaagccgca cgttatgaag atgaaagcct gaacctgggc gattacgttg aagatcagat
     5041 tgagtctgtt acctttgacc gtatcactac ccagacggca aaacaggtta tcgtgcagaa
     5101 agtgcgtgaa gccgaacgtg cgatggtggt tgatcagttc cgtgaacacg aaggtgaaat
     5161 catcaccggc gtggtgaaaa aagtaaaccg cgacaacatc tctctggatc tgggcaacaa
     5221 cgctgaagcc gtgatcctgc gcgaagatat gctgccgcgt gaaaacttcc gccctggcga
     5281 ccgcgttcgt ggcgtgctct attccgttcg cccggaagcg cgtggcgcgc aactgttcgt
     5341 cactcgttcc aagccggaaa tgctgatcga actgttccgt attgaagtgc cagaaatcgg
     5401 cgaagaagtg attgaaatta aagcagcggc tcgcgatccg ggttctcgtg cgaaaatcgc
     5461 ggtgaaaacc aacgataaac gtatcgatcc ggtaggtgct tgcgtaggta tgcgtggcgc
     5521 gcgtgttcag gcggtgtcta ctgaactggg tggcgagcgt atcgatatcg tcctgtggga
     5581 tgataacccg gcgcagttcg tgattaacgc aatggcaccg gcagacgttg cttctatcgt
     5641 ggtggatgaa gataaacaca ctatggacat cgccgttgaa gccggtaatc tggcgcaggc
     5701 gattggccgt aacggtcaga acgtgcgtct ggcttcgcaa ctgagcggtt gggaactcaa
     5761 cgtgatgacc gttgacgacc tgcaagctaa gcatcaggcg gaagcgcacg cagcgatcga
     5821 caccttcacc aaatatctcg acatcgacga agacttcgcg actgttctgg tagaagaagg
     5881 cttctcgacg ctggaagaat tggcctatgt gccgatgaaa gagctgttgg aaatcgaagg
     5941 ccttgatgag ccgaccgttg aagcactgcg cgagcgtgct aaaaatgcac tggccaccat
     6001 tgcacaggcc caggaagaaa gcctcggtga taacaaaccg gctgacgatc tgctgaacct
     6061 tgaaggggta gatcgtgatt tggcattcaa actggccgcc cgtggcgttt gtacgctgga
     6121 agatctcgcc gaacagggca ttgatgatct ggctgatatc gaagggttga ccgacgaaaa
     6181 agccggagca ctgattatgg ctgcccgtaa tatttgctgg ttcggtgacg aagcgactag
     6241 tggttctcgc ggccgcgatg tttccgagct gtgcaagctg aagaaagagc ttgaggccaa
     6301 ggctagggtt aagcagatat gcaccagcaa gttggaggac attctcaagt tcgtggacac
     6361 gaaaatatac atcgaggtgc gaccgcgcta tcttctaccg gataccaact gctttatcga
     6421 ctgccttgaa gactttgaaa aactgtccac ggagtttaaa cgctacacac ttattatacc
     6481 actgacagtg gttaaggaac tggatggcct ctccaagggc gttaagttgg actcgtaccg
     6541 cagttcgaag cagacgcagc gcatccacca ctttgacgag gtgtcgtcgc gggccaaaaa
     6601 gtcgttagag ttcatcaagt cggccaaagg caatgtgaaa tgtgctacca caaagggctc
     6661 gtttgtcaat gcatcggtgt ttgccctagt tgaggaggaa tatttgtcca atgacgacaa
     6721 gattctggca acggcaatgg ctgtctcgaa aacggcaaaa accgaacaat gcacagcaga
     6781 tggcaagtgt ttcattcaaa cggaattggt gctcattaca accgatcgta atctgcgtgt
     6841 gaaggctctt tctcgcaatt tggccgtcag cgcgttggat gagtttctgc aatgggccaa
     6901 ggattgccgc gaatcaaccg gaggacgctc gagtctagag ggcccgcggt tcgaaggtaa
     6961 gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggtcatc atcaccatca
     7021 ccattgagtt taaacccgct gatcagcctc gactgtgcct tctaaggcct gagctcgctg
     7081 atcagcctcg atcgaggatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     7141 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     7201 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     7261 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     7321 tatggctgat tatgatcagt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
     7381 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     7441 aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac cgctttcgct
     7501 ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc
     7561 aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc attatcgccg
     7621 gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga ggctggatgg
     7681 ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca
     7741 tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg ctcgcggctc
     7801 ttaccagcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7861 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7921 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7981 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     8041 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     8101 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     8161 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     8221 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     8281 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     8341 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     8401 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     8461 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     8521 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     8581 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     8641 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     8701 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     8761 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     8821 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     8881 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg
     8941 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     9001 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     9061 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     9121 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     9181 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata
     9241 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     9301 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     9361 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     9421 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     9481 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     9541 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     9601 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     9661 ggccctttcg t
//
