LOCUS       sequence-289034-        8927 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian Expression of HsSmg6-mut-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8927)
  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 8927)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pEYFP-N1-HsSmg6-mut-siRNAres_I
FEATURES             Location/Qualifiers
     source          1..8927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4867..4876
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4873..4875,4876..4878,4879..5592)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4918..4939)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5179..5198)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5526..5547
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5715..5836
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5752..5771)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5806..5825
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5843..6298)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5980..6001)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6192..6211
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6325..6429
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6431..6788
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6431..6622
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6454..6474)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6639..6774
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6823..7617
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6823..7617
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6877..6896)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7487..7506
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7805..7824)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7849..7896
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8225..8813
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagccaac
      601 atggcggaag ggctggagcg tgtgcggatc tccgcgtcgg agctgcgcgg gatcctggct
      661 actctggccc cgcaggccgg gagcagagaa aacatgaagg aattaaagga ggccaggccg
      721 cgcaaagata acaggcgtcc agatctggaa atctataagc ctggcctttc tcggctaagg
      781 aacaagccca aaatcaagga accccctggg agtgaggaat tcaaagatga aattgttaat
      841 gaccgagatt gctctgctgt tgaaaatggt acacagcccg ttaaagatgt ctgcaaggaa
      901 ctgaacaacc aagagcagaa tggtcctata gacccagaaa ataatcgggg acaagaatcc
      961 tttcctagga ctgctggaca agaggatcgt agtctaaaaa ttatcaaaag aacaaagaaa
     1021 cccgacctgc agatctatca gcctggacga cgtttgcaga ctgttagcaa agaatccgcc
     1081 agtcgggtgg aggaggaaga agtcctcaac caggtagaac aactgagagt agaggaagat
     1141 gagtgtaggg gaaatgttgc gaaggaggaa gttgcgaata aaccagacag ggccgagata
     1201 gaaaagagcc caggtggtgg gagagtaggg gctgcaaaag gagaaaaagg aaagaggatg
     1261 ggaaaagggg agggggtgag ggaaacccac gacgacccgg cccgcgggag gccgggctcc
     1321 gcaaagcgct actcccgctc agacaaacga aggaatcgct accgcacgcg cagcaccagc
     1381 tcagctggca gcaacaacag cgctgaggga gctggcctga cggataatgg atgtcgccgc
     1441 cgccgacagg ataggaccaa ggagaggcca ccactgaaga agcaagtgtc tgtgtcctca
     1501 accgattcct tagacgagga cagaattgat gagcctgatg gattaggacc caggagaagt
     1561 tcagaaagga agagacattt agaaagaaac tggtctggcc gtggggaggg tgagcagaaa
     1621 accagtgcta aagaatatcg aggcactctt cgtgtcactt tcgatgcaga agccatgaac
     1681 aaagagtctc ccatggtgag gtcagccagg gatgatatgg atagaggaaa gcctgacaaa
     1741 ggcttgagca gtgggggcaa aggctctgag aagcaggagt ccaaaaaccc gaaacaagaa
     1801 cttcggggtc gtggtcgtgg cattctgatt ttgcctgccc ataccaccct atctgtcaat
     1861 tcagcaggtt ctccagagtc cgcgcctttg ggacctcggc ttttgtttgg atctggtagt
     1921 aagggatctc ggagttgggg ccgtggaggc accacacgcc gattgtggga cccaaacaat
     1981 cctgatcaga aacctgctct aaagactcag acgccccagc tacatttctt ggacactgat
     2041 gatgaagtca gccctacatc ttggggtgac tcacgccagg ctcaggcatc ttactataag
     2101 tttcaaaact ctgacaaccc ctattattac ccccggacac caggccctgc ctcccagtat
     2161 ccctatacgg gctataaccc tctacagtac ccagtgggcc ctacgaatgg tgtgtaccca
     2221 gggccttact acccaggcta cccgactccg tcaggacagt atgtgtgtag ccctctacct
     2281 accagcacca tgagtcccga ggaggtagag cagcacatga ggaacctgca gcaacaggag
     2341 ctgcacaggc ttctccgggt ggctgacaac caggaactgc agctcagcaa cctgctctcc
     2401 agggaccgca tcagtccgga gggcctggag aagatggcgc aactcagagc tgaactgctg
     2461 cagctatatg agcgctgtat tctattagat attgagttct ctgataatca gaatgtggat
     2521 cagatcctgt ggaagaatgc tttctatcag gtgattgaga agttcaggca acttgtcaag
     2581 gatccgaatg ttgagaaccc agaacagatt cggaacagac ttttggagct cttggatgag
     2641 ggtagtgact tctttgatag tttgcttcag aagctgcagg ttacttacaa gttcaaactg
     2701 gaagactaca tggatggtct tgccattcgc agcaagccat tacgcaagac agtaaaatat
     2761 gccttgatca gtgcccagcg atgcatgata tgccaaggag atattgctag gtaccgggag
     2821 caggcaagcg acacggcaaa ctatgggaaa gcacgcagtt ggtacctgaa ggcccagcac
     2881 attgctccca agaatgggcg cccctataac cagttggctt tgctggcagt gtatacgagg
     2941 aggaagcttg acgctgtcta ttactatatg cgcagtttag ctgccagcaa ccctatcctg
     3001 actgccaagg agagtctcat gagcttgttt gaagagacca agcggaaggc agaacagatg
     3061 gaaaagaagc aacatgagga atttgacctg agccctgacc agtggcggaa aggaaagaag
     3121 tctactttcc ggcatgttgg agatgacacc actcgcctgg agatctggat tcatccatcc
     3181 catccacggt cttcccaggg cactgagtct gggaaggatt ctgagcaaga gaatgggctg
     3241 ggcagcctga gtcccagtga tctgaacaaa aggttcatcc tcagttttct ccatgcccat
     3301 gggaagctgt ttacccggat tgggatggag acattccctg cagtggctga gaaggtcctc
     3361 aaggagttcc aggtgttact gcagcacagc ccctctccca ttggaagtac ccgcatgctg
     3421 cagcttatga ccatcaatat gtttgcagta cacaactccc agctgaaaga ctgcttctcg
     3481 gaggagtgcc gctctgtgat ccaggaacaa gccgcagctc tgggcttggc catgttttct
     3541 ctactggtcc gccgctgcac ctgcttactt aaggagtccg ccaaagctca gctgtcctct
     3601 cctgaggacc aggatgacca agacgacatc aaggtgtctt cctttgtccc ggacctgaag
     3661 gagctgctcc ccagtgtcaa agtctggtca gattggatgc tcggctaccc ggacacctgg
     3721 aatcctcctc ccacatccct ggatctgccc tcgcatgttg ctgtggatgt atggtcgacg
     3781 ctggctgatt tctgtaacat actgactgca gtgaatcagt ctgaggtgcc actgtacaag
     3841 gacccggatg atgacctcac ccttcttatc ctggaagagg atcggcttct ctcgggcttt
     3901 gtccccttgc tggctgcccc tcaggacccc tgctacgtgg agaaaacctc ggataaggtt
     3961 attgcagctg actgcaaaag ggtcacagtg ctgaagtatt ttctggaagc cctttgtgga
     4021 caagaagagc ctctgctggc attcaagggt ggaaagtatg tgtcagtggc acccgtccca
     4081 gacaccatgg gaaaggaaat gggaagccaa gagggaacac gactggaaga tgaggaggag
     4141 gatgtggtga ttgaagactt tgaggaagat tcagaggctg aaggcagcgg aggcgaggat
     4201 gacatcaggg agcttcgggc caagaagctg gctctggcca ggaagatagc tgagcagcag
     4261 cgtcgccagg aaaagatcca ggctgtcctg gaggaccaca gtcagatgag gcagatggag
     4321 ctcgaaatca gacctttgtt cctcgtacca gctaccaacg gcttcattga ccacctggcc
     4381 agtctggcgc ggctgctgga gagcaggaag tacatcctgg tggtgcccct catcgtgatc
     4441 aatgagctgg acggcctggc caaggggcag gagacagacc accgggctgg gggctacgcc
     4501 cgtgtggtac aagagaaggc ccgcaagtcc atcgagttcc tcgagcagcg attcgagagt
     4561 cgggactctt gcctgcgagc cctgaccagc cgtggcaatg aactcgaatc catcgccttc
     4621 cgcagtgagg acatcactgg ccagctgggt aacaacgatg atctcatcct gtcctgctgc
     4681 ctccactact gcaaagacaa ggctaaggac ttcatgcccg ccagcaaaga ggagccaatc
     4741 cggctactgc gggaggtggt gctgttgacg gatgctcgga acctgcgtgt gaaggcgctc
     4801 acaaggaatg ttcctgtacg ggacatccca gccttcctca cgtgggccca ggtgggcaaa
     4861 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     4921 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     4981 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     5041 ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc
     5101 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     5161 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     5221 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     5281 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     5341 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     5401 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     5461 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     5521 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     5581 ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     5641 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     5701 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5761 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5821 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     5881 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     5941 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     6001 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     6061 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     6121 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     6181 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     6241 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     6301 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6361 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6421 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     6481 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     6541 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     6601 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     6661 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6721 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     6781 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     6841 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     6901 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     6961 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     7021 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     7081 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     7141 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     7201 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     7261 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     7321 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     7381 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     7441 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     7501 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     7561 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     7621 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     7681 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     7741 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     7801 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     7861 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     7921 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     7981 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     8041 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     8101 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     8161 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     8221 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     8281 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     8341 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     8401 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     8461 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     8521 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     8581 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     8641 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     8701 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     8761 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     8821 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     8881 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
