LOCUS       sequence-357455-        8290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg5-G120E-del133-155.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8290)
  AUTHORS   Loh B, Jonas S, Izaurralde E
  TITLE     The SMG5-SMG7 heterodimer directly recruits the CCR4-NOT deadenylase
            complex to mRNAs containing nonsense codons via interaction with 
            POP2.
  JOURNAL   Genes Dev. 2013 Oct 1;27(19):2125-38. doi: 10.1101/gad.226951.113.
  PUBMED    24115769
REFERENCE   2  (bases 1 to 8290)
  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"; doi: 
            "10.1101/gad.226951"; journalName: "Genes Dev"; date: "2013-10-1- 
            1"; volume: "27"; issue: "19"; pages: "2125-38"
COMMENT     Sequence Label: pN1-HsSmg5-G120E-del133-155-V5-SBP-MBP_V
FEATURES             Location/Qualifiers
     source          1..8290
                     /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"
     CDS             3661..3702
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3724..3837
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWGGGHVVEGLAGELEQLRARLEHHPQGQREP"
     regulatory      4819..4828
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5078..5199
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5115..5134)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5169..5188
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5206..5661)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5343..5364)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5555..5574
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5688..5792
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5794..6151
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5794..5985
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5817..5837)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6002..6137
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6186..6980
                     /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             6186..6980
                     /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(6240..6259)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6850..6869
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7168..7187)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7212..7259
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7588..8176
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8077..8096
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
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 gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcgcaaca tgagccaagg cccccccaca
      661 ggggagagca gcgagcccga agcaaaagtc ctccacacta agcggcttta ccgggctgtg
      721 gtggaggctg tgcatcgact tgacctcatc ctttgcaaca aaactgctta tcaagaagta
      781 ttcaaaccag aaaacattag cctgaggaac aagctgcgtg agctctgcgt caagcttatg
      841 ttcctgcacc cagtggacta tgggagaaag gctgaggagc tgctgtggag aaaggtatac
      901 tatgaagtta tccagcttat caagactaac aaaaagcaca tccacagccg gagcactttg
      961 gaatgtgcct acaggacgca cctggttgct ggtattgagt tctaccagca tctccttctc
     1021 tatatccagt cccacccagt gtctgcctca gggaaggaga tggattgggc acagatggca
     1081 tgtcaccgat gtctggtgta tctgggggat ttgtcccgat atcagaatga attagctggc
     1141 gtagataccg agctgctagc cgagagattt tactaccaag ccctgtcagt agctcctcag
     1201 attggaatgc ccttcaatca gctgggcacc ctggcaggca gcaagtacta taatgtggaa
     1261 gccatgtatt gctacctgcg ctgcatccag tcagaagtgt cctttgaggg agcctatggg
     1321 aacctcaagc ggctgtatga caaggcagcc aaaatgtacc accaactgaa gaagtgtgag
     1381 actcggaaac tgtctcctgg caaaaagcga tgtaaagaca ttaaaaggtt gctagtgaac
     1441 tttatgtatc tgcaaagcct cctacagccc aaaagcagct ccgtggactc agagctgacc
     1501 tcactttgcc agtcagtcct ggaggacttc aacctctgtc tcttctacct gccctcctca
     1561 cccaacctca gcctggccag tgaggatgag gaggagtatg agagtggata tgctttcctc
     1621 ccggaccttc tcatctttca aatggtcatc atctgcctta tgtgtgtgca cagcttggag
     1681 agagcaggat ccaagcagta cagtgcagcc attgccttca ccctggccct cttttcccac
     1741 ctcgtcaatc atgtcaacat acggctgcag gctgagctgg aagagggcga gaatcccgtc
     1801 ccggcattcc agagtgatgg cacagatgaa ccagagtcca aggaacctgt ggagaaagag
     1861 gaggagccag atcctgagcc tcctcctgta acaccccaag tgggtgaggg cagaaagagc
     1921 cgtaagttct ctcgcctctc ctgtctccgc cgtcgccgcc acccacccaa agttggtgat
     1981 gacagtgacc tgagtgaagg ctttgaatcg gactcaagcc atgactcagc ccgggccagt
     2041 gagggctcag acagtggctc tgacaagagt cttgaaggtg ggggaacggc ctttgatgct
     2101 gaaacagact cggaaatgaa tagccaggag tcccgatcag acttggaaga tatggaggaa
     2161 gaggagggga cacggtcacc aaccctggag ccccctcggg gcagatcaga ggctcccgat
     2221 tccctcaatg gcccactggg ccccagtgag gctagcattg ccagcaatct acaagccatg
     2281 tccacccaga tgttccagac taagcgctgc ttccgactgg cccccacctt tagcaacctg
     2341 ctcctccagc ccaccaccaa ccctcatacc tcggccagcc acaggccttg cgtcaatggg
     2401 gatgtagaca agccttcaga gccagcctct gaggagggct ctgagtcgga ggggagtgag
     2461 tccagtggac gctcctgtcg gaatgagcgc agcatccagg agaagcttca ggtcctgatg
     2521 gccgaaggtc tgcttcctgc tgtgaaagtc ttcctggact ggcttcggac caaccccgac
     2581 ctcatcatcg tgtgtgcgca gagctctcaa agtctgtgga accgcctgtc tgtgttgctg
     2641 aatctgttgc ctgctgctgg tgaactccag gagtctggcc tggccttgtg tcctgaggtc
     2701 caagatcttc ttgaaggttg tgaactgcct gacctcccct ctagccttct gctcccagag
     2761 gacatggctc ttcgtaacct gcccccgctc cgagctgccc acagacgctt taactttgac
     2821 acggatcggc ccctgctcag caccttagag gagtcagtgg tgcgcatctg ctgcatccgc
     2881 agctttggtc atttcatcgc ccgcctgcaa ggcagcatcc tgcagttcaa cccagaggtt
     2941 ggcatcttcg tcagcattgc ccagtctgag caggagagcc tgctgcagca ggcccaggca
     3001 cagttccgaa tggcacagga ggaagctcgt cggaacaggc tcatgagaga catggctcag
     3061 ctacgacttc agctcgaagt gtctcagctg gagggcagcc tgcagcagcc caaggcccag
     3121 tcagccatgt ctccctacct cgtccctgac acccaggccc tctgccacca tctccctgtc
     3181 atccgccaac tggccaccag tggccgcttc attgtcatca tcccaaggac agtgatcgat
     3241 ggcctggatt tgctgaagaa ggaacaccca ggggcccggg atgggattcg gtacctggag
     3301 gcagagttta aaaaaggaaa caggtacatt cgctgccaga aagaggtggg aaagagcttt
     3361 gagcggcata agctgaagag gcaggatgca gatgcctgga ctctctataa gatcctagac
     3421 agctgcaaac agctgactct ggcccagggg gcaggtgagg aggatccgag tggcatggtg
     3481 accatcatca caggccttcc actggacaac cccagcgtgc tttcaggccc catgcaggca
     3541 gccctgcagg ccgctgccca cgccagtgtg gacatcaaga atgttctgga cttctacaag
     3601 cagtggaagg aaattggtac gtcgacggta ccgcgggccc gggatccacc ggtccctcca
     3661 ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgcccggaag cggtagcggt
     3721 accatggacg agaagaccac cggctggggg ggcggccacg tggtggaggg cctggccggc
     3781 gagctggagc agctgcgggc caggctggag caccaccctc agggccagcg ggagccctcc
     3841 ggcggctgca agcccctcga catgaaaatc gaagaaggta aactggtaat ctggattaac
     3901 ggcgataaag gctataacgg tctcgctgaa gtcggtaaga aattcgagaa agataccgga
     3961 attaaagtca ccgttgagca tccggataaa ctggaagaga aattcccgca ggttgcggca
     4021 actggcgatg gccctgacat tatcttctgg gcacacgacc gctttggtgg ctacgctcaa
     4081 tctggcctgt tggctgaaat caccccggac aaagcgttcc aggacaagct gtatccgttt
     4141 acctgggatg ccgtacgtta caacggcaag ctgattgctt acccgatcgc tgttgaagcg
     4201 ttatcgctga tttataacaa agatctgctg ccgaacccgc caaaaacctg ggaagagatc
     4261 ccggcgctgg ataaagaact gaaagcgaaa ggtaagagcg cgctgatgtt caacctgcaa
     4321 gaaccgtact tcacctggcc gctgattgct gctgacgggg gttatgcgtt caagtatgaa
     4381 aacggcaagt acgacattaa agacgtgggc gtggataacg ctggcgcgaa agcgggtctg
     4441 accttcctgg ttgacctgat taaaagcaaa cacatgaatg cagacaccga ttactccatc
     4501 gcagaagctg cctttaataa aggcgaaaca gcgatgacca tcaacggccc gtgggcatgg
     4561 tccaacatcg acaccagcaa agtgaattat ggtgtaacgg tactgccgac cttcaagggt
     4621 caaccatcca aaccgttcgt tggcgtgctg agcgcaggta ttaacgccgc cagtccgaac
     4681 aaagagctgg cgaaagagtt cctcgaaaac tatctgctga ctgatgaagg tctggaagcg
     4741 gttaataaag acaaaccgct gggtgccgta gcgctgaagt cttacgagga agagttggcg
     4801 aaagatccac gtattgccgc caccatggaa aacgcccaga aaggtgaaat catgccgaac
     4861 atcccgcaga tgtccgcttt ctggtatgcc gtgcgtactg cggtgatcaa cgccgccagc
     4921 ggtcgtcaga ctgtcgatga agccctgaaa gactaagcgg ccgcgactct agatcataat
     4981 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     5041 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     5101 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5161 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     5221 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     5281 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     5341 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     5401 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     5461 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     5521 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     5581 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     5641 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5701 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5761 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     5821 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5881 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     5941 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     6001 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     6061 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     6121 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     6181 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     6241 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     6301 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     6361 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     6421 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     6481 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     6541 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     6601 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     6661 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     6721 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     6781 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     6841 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     6901 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6961 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     7021 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     7081 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     7141 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     7201 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     7261 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     7321 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     7381 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     7441 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     7501 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     7561 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     7621 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     7681 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     7741 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     7801 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     7861 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     7921 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7981 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     8041 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     8101 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     8161 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     8221 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     8281 cgccatgcat
//
