LOCUS       sequence-357487-        8919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg5-G120E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8919)
  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 8919)
  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: pcDNA3.1-MS2-HA-HsSmg5-G120E_W
FEATURES             Location/Qualifiers
     source          1..8919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1333..1359
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(4472..4490)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4473..4490)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4510..4527)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4516..4740
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4786..5214
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4873..4892)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5083..5104
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5223..5243)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5228..5557
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5408..5543
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5470..5489
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5624..6418
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5678..5697)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6288..6307
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6592..6713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6629..6648)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6683..6702
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6762..6778)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6762..6778)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6775..6797)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6786..6802
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6810..6816,6817..6834,6835..6840))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6855..6876
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6993..7010)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7164..7752)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7244..7263)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7923..8714,8715..8783))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8546..8565
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8784..8888)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
      901 atggcttcta actttactca gttcgttctc gtcgacaatg gcggaactgg cgacgtgact
      961 gtcgccccaa gcaacttcgc taacggggtc gctgaatgga tcagctctaa ctcgcgatca
     1021 caggcttaca aagtaacctg tagcgttcgt cagagctctg cgcagaatcg caaatacacc
     1081 atcaaagtcg aggtgcctaa agtggcaacc cagactgttg gtggtgaaga gcttcctgta
     1141 gccggatgga gatcttactt aaatatggaa ctaaccattc caattttcgc cacgaattcc
     1201 gactgcgagc ttattgttaa ggcaatgcaa ggtctcctaa aagatggaaa cccgattccc
     1261 tcggccatcg cggccaactc cggcatctac ggaggtggag gtggatctgg tggatctcct
     1321 ccacatatgg cttacccata cgatgttcca gattacgctc ctccaaagct tggatccgaa
     1381 ttcccgggtc gactcgagat gagccaaggc ccccccacag gggagagcag cgagcccgaa
     1441 gcaaaagtcc tccacactaa gcggctttac cgggctgtgg tggaggctgt gcatcgactt
     1501 gacctcatcc tttgcaacaa aactgcttat caagaagtat tcaaaccaga aaacattagc
     1561 ctgaggaaca agctgcgtga gctctgcgtc aagcttatgt tcctgcaccc agtggactat
     1621 gggagaaagg ctgaggagct gctgtggaga aaggtatact atgaagttat ccagcttatc
     1681 aagactaaca aaaagcacat ccacagccgg agcactttgg aatgtgccta caggacgcac
     1741 ctggttgctg gtattgagtt ctaccagcat ctccttctct atatccagtc ccactaccag
     1801 ctggaactgc agtgctgcat cgactggacc catgtcactg accccctcat aggatgcaag
     1861 aagccagtgt ctgcctcagg gaaggagatg gattgggcac agatggcatg tcaccgatgt
     1921 ctggtgtatc tgggggattt gtcccgatat cagaatgaat tagctggcgt agataccgag
     1981 ctgctagccg agagatttta ctaccaagcc ctgtcagtag ctcctcagat tggaatgccc
     2041 ttcaatcagc tgggcaccct ggcaggcagc aagtactata atgtggaagc catgtattgc
     2101 tacctgcgct gcatccagtc agaagtgtcc tttgagggag cctatgggaa cctcaagcgg
     2161 ctgtatgaca aggcagccaa aatgtaccac caactgaaga agtgtgagac tcggaaactg
     2221 tctcctggca aaaagcgatg taaagacatt aaaaggttgc tagtgaactt tatgtatctg
     2281 caaagcctcc tacagcccaa aagcagctcc gtggactcag agctgacctc actttgccag
     2341 tcagtcctgg aggacttcaa cctctgtctc ttctacctgc cctcctcacc caacctcagc
     2401 ctggccagtg aggatgagga ggagtatgag agtggatatg ctttcctccc ggaccttctc
     2461 atctttcaaa tggtcatcat ctgccttatg tgtgtgcaca gcttggagag agcaggatcc
     2521 aagcagtaca gtgcagccat tgccttcacc ctggccctct tttcccacct cgtcaatcat
     2581 gtcaacatac ggctgcaggc tgagctggaa gagggcgaga atcccgtccc ggcattccag
     2641 agtgatggca cagatgaacc agagtccaag gaacctgtgg agaaagagga ggagccagat
     2701 cctgagcctc ctcctgtaac accccaagtg ggtgagggca gaaagagccg taagttctct
     2761 cgcctctcct gtctccgccg tcgccgccac ccacccaaag ttggtgatga cagtgacctg
     2821 agtgaaggct ttgaatcgga ctcaagccat gactcagccc gggccagtga gggctcagac
     2881 agtggctctg acaagagtct tgaaggtggg ggaacggcct ttgatgctga aacagactcg
     2941 gaaatgaata gccaggagtc ccgatcagac ttggaagata tggaggaaga ggaggggaca
     3001 cggtcaccaa ccctggagcc ccctcggggc agatcagagg ctcccgattc cctcaatggc
     3061 ccactgggcc ccagtgaggc tagcattgcc agcaatctac aagccatgtc cacccagatg
     3121 ttccagacta agcgctgctt ccgactggcc cccaccttta gcaacctgct cctccagccc
     3181 accaccaacc ctcatacctc ggccagccac aggccttgcg tcaatgggga tgtagacaag
     3241 ccttcagagc cagcctctga ggagggctct gagtcggagg ggagtgagtc cagtggacgc
     3301 tcctgtcgga atgagcgcag catccaggag aagcttcagg tcctgatggc cgaaggtctg
     3361 cttcctgctg tgaaagtctt cctggactgg cttcggacca accccgacct catcatcgtg
     3421 tgtgcgcaga gctctcaaag tctgtggaac cgcctgtctg tgttgctgaa tctgttgcct
     3481 gctgctggtg aactccagga gtctggcctg gccttgtgtc ctgaggtcca agatcttctt
     3541 gaaggttgtg aactgcctga cctcccctct agccttctgc tcccagagga catggctctt
     3601 cgtaacctgc ccccgctccg agctgcccac agacgcttta actttgacac ggatcggccc
     3661 ctgctcagca ccttagagga gtcagtggtg cgcatctgct gcatccgcag ctttggtcat
     3721 ttcatcgccc gcctgcaagg cagcatcctg cagttcaacc cagaggttgg catcttcgtc
     3781 agcattgccc agtctgagca ggagagcctg ctgcagcagg cccaggcaca gttccgaatg
     3841 gcacaggagg aagctcgtcg gaacaggctc atgagagaca tggctcagct acgacttcag
     3901 ctcgaagtgt ctcagctgga gggcagcctg cagcagccca aggcccagtc agccatgtct
     3961 ccctacctcg tccctgacac ccaggccctc tgccaccatc tccctgtcat ccgccaactg
     4021 gccaccagtg gccgcttcat tgtcatcatc ccaaggacag tgatcgatgg cctggatttg
     4081 ctgaagaagg aacacccagg ggcccgggat gggattcggt acctggaggc agagtttaaa
     4141 aaaggaaaca ggtacattcg ctgccagaaa gaggtgggaa agagctttga gcggcataag
     4201 ctgaagaggc aggatgcaga tgcctggact ctctataaga tcctagacag ctgcaaacag
     4261 ctgactctgg cccagggggc aggtgaggag gatccgagtg gcatggtgac catcatcaca
     4321 ggccttccac tggacaaccc cagcgtgctt tcaggcccca tgcaggcagc cctgcaggcc
     4381 gctgcccacg ccagtgtgga catcaagaat gttctggact tctacaagca gtggaaggaa
     4441 attggttagg cggccgccta gagggcccta ttctatagtg tcacctaaat gctagagctc
     4501 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     4561 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     4621 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     4681 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     4741 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     4801 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4861 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4921 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4981 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     5041 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     5101 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     5161 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     5221 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5281 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     5341 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     5401 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     5461 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     5521 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     5581 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     5641 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     5701 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     5761 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     5821 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5881 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5941 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     6001 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     6061 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     6121 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     6181 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     6241 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     6301 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     6361 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     6421 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     6481 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     6541 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     6601 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     6661 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     6721 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     6781 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6841 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6901 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6961 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     7021 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7081 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     7141 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7201 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7261 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7321 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7381 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7441 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7501 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7561 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     7621 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7681 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7741 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7801 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7861 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7921 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7981 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     8041 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     8101 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     8161 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     8221 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     8281 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     8341 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8401 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8461 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8521 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8581 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8641 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8701 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8761 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8821 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8881 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
