LOCUS       sequence-290213-        7849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg5-del847-1017.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7849)
  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 7849)
  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-del847-1017-V5-SBP-MBP_V
FEATURES             Location/Qualifiers
     source          1..7849
                     /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             3220..3261
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3283..3396
                     /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      4378..4387
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4637..4758
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4674..4693)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4728..4747
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4765..5220)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4902..4923)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5114..5133
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5247..5351
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5353..5710
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5353..5544
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5376..5396)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5561..5696
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5745..6539
                     /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             5745..6539
                     /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(5799..5818)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6409..6428
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6727..6746)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6771..6818
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7147..7735
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7636..7655
                     /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 ggtattggct tctaccagca tctccttctc
     1021 tatatccagt cccactacca gctggaactg cagtgctgca tcgactggac ccatgtcact
     1081 gaccccctca taggatgcaa gaagccagtg tctgcctcag ggaaggagat ggattgggca
     1141 cagatggcat gtcaccgatg tctggtgtat ctgggggatt tgtcccgata tcagaatgaa
     1201 ttagctggcg tagataccga gctgctagcc gagagatttt actaccaagc cctgtcagta
     1261 gctcctcaga ttggaatgcc cttcaatcag ctgggcaccc tggcaggcag caagtactat
     1321 aatgtggaag ccatgtattg ctacctgcgc tgcatccagt cagaagtgtc ctttgaggga
     1381 gcctatggga acctcaagcg gctgtatgac aaggcagcca aaatgtacca ccaactgaag
     1441 aagtgtgaga ctcggaaact gtctcctggc aaaaagcgat gtaaagacat taaaaggttg
     1501 ctagtgaact ttatgtatct gcaaagcctc ctacagccca aaagcagctc cgtggactca
     1561 gagctgacct cactttgcca gtcagtcctg gaggacttca acctctgtct cttctacctg
     1621 ccctcctcac ccaacctcag cctggccagt gaggatgagg aggagtatga gagtggatat
     1681 gctttcctcc cggaccttct catctttcaa atggtcatca tctgccttat gtgtgtgcac
     1741 agcttggaga gagcaggatc caagcagtac agtgcagcca ttgccttcac cctggccctc
     1801 ttttcccacc tcgtcaatca tgtcaacata cggctgcagg ctgagctgga agagggcgag
     1861 aatcccgtcc cggcattcca gagtgatggc acagatgaac cagagtccaa ggaacctgtg
     1921 gagaaagagg aggagccaga tcctgagcct cctcctgtaa caccccaagt gggtgagggc
     1981 agaaagagcc gtaagttctc tcgcctctcc tgtctccgcc gtcgccgcca cccacccaaa
     2041 gttggtgatg acagtgacct gagtgaaggc tttgaatcgg actcaagcca tgactcagcc
     2101 cgggccagtg agggctcaga cagtggctct gacaagagtc ttgaaggtgg gggaacggcc
     2161 tttgatgctg aaacagactc ggaaatgaat agccaggagt cccgatcaga cttggaagat
     2221 atggaggaag aggaggggac acggtcacca accctggagc cccctcgggg cagatcagag
     2281 gctcccgatt ccctcaatgg cccactgggc cccagtgagg ctagcattgc cagcaatcta
     2341 caagccatgt ccacccagat gttccagact aagcgctgct tccgactggc ccccaccttt
     2401 agcaacctgc tcctccagcc caccaccaac cctcatacct cggccagcca caggccttgc
     2461 gtcaatgggg atgtagacaa gccttcagag ccagcctctg aggagggctc tgagtcggag
     2521 gggagtgagt ccagtggacg ctcctgtcgg aatgagcgca gcatccagga gaagcttcag
     2581 gtcctgatgg ccgaaggtct gcttcctgct gtgaaagtct tcctggactg gcttcggacc
     2641 aaccccgacc tcatcatcgt gtgtgcgcag agctctcaaa gtctgtggaa ccgcctgtct
     2701 gtgttgctga atctgttgcc tgctgctggt gaactccagg agtctggcct ggccttgtgt
     2761 cctgaggtcc aagatcttct tgaaggttgt gaactgcctg acctcccctc tagccttctg
     2821 ctcccagagg acatggctct tcgtaacctg cccccgctcc gagctgccca cagacgcttt
     2881 aactttgaca cggatcggcc cctgctcagc accttagagg agtcagtggt gcgcatctgc
     2941 tgcatccgca gctttggtca tttcatcgcc cgcctgcaag gcagcatcct gcagttcaac
     3001 ccagaggttg gcatcttcgt cagcattgcc cagtctgagc aggagagcct gctgcagcag
     3061 gcccaggcac agttccgaat ggcacaggag gaagctcgtc ggaacaggct catgagagac
     3121 atggctcagc tacgacttca gctcgaagtg tctcagctgg agggcagcct gcagcagacg
     3181 tcgacggtac cgcgggcccg ggatccaccg gtccctccag gtaagcctat ccctaaccct
     3241 ctcctcggtc tcgattctac gcccggaagc ggtagcggta ccatggacga gaagaccacc
     3301 ggctgggggg gcggccacgt ggtggagggc ctggccggcg agctggagca gctgcgggcc
     3361 aggctggagc accaccctca gggccagcgg gagccctccg gcggctgcaa gcccctcgac
     3421 atgaaaatcg aagaaggtaa actggtaatc tggattaacg gcgataaagg ctataacggt
     3481 ctcgctgaag tcggtaagaa attcgagaaa gataccggaa ttaaagtcac cgttgagcat
     3541 ccggataaac tggaagagaa attcccgcag gttgcggcaa ctggcgatgg ccctgacatt
     3601 atcttctggg cacacgaccg ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc
     3661 accccggaca aagcgttcca ggacaagctg tatccgttta cctgggatgc cgtacgttac
     3721 aacggcaagc tgattgctta cccgatcgct gttgaagcgt tatcgctgat ttataacaaa
     3781 gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga taaagaactg
     3841 aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag aaccgtactt cacctggccg
     3901 ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa acggcaagta cgacattaaa
     3961 gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt
     4021 aaaagcaaac acatgaatgc agacaccgat tactccatcg cagaagctgc ctttaataaa
     4081 ggcgaaacag cgatgaccat caacggcccg tgggcatggt ccaacatcga caccagcaaa
     4141 gtgaattatg gtgtaacggt actgccgacc ttcaagggtc aaccatccaa accgttcgtt
     4201 ggcgtgctga gcgcaggtat taacgccgcc agtccgaaca aagagctggc gaaagagttc
     4261 ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg
     4321 ggtgccgtag cgctgaagtc ttacgaggaa gagttggcga aagatccacg tattgccgcc
     4381 accatggaaa acgcccagaa aggtgaaatc atgccgaaca tcccgcagat gtccgctttc
     4441 tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa
     4501 gccctgaaag actaagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     4561 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     4621 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4681 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4741 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     4801 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     4861 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4921 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4981 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     5041 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     5101 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     5161 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     5221 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5281 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5341 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     5401 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5461 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5521 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     5581 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5641 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5701 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     5761 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5821 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5881 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     5941 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     6001 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     6061 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     6121 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     6181 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     6241 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     6301 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     6361 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     6421 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     6481 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     6541 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     6601 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     6661 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     6721 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     6781 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     6841 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     6901 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     6961 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     7021 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7081 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7141 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7201 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7261 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     7321 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7381 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7441 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7501 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7561 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7621 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7681 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7741 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     7801 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
