LOCUS       sequence-290001-        8847 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg5-del133-155-G120E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8847)
  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 8847)
  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-del133-155-G120E_T
FEATURES             Location/Qualifiers
     source          1..8847
                     /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(4403..4421)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4404..4421)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4441..4458)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4447..4671
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4717..5145
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4804..4823)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5014..5035
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5154..5174)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5159..5488
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5339..5474
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5401..5420
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5555..6349
                     /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(5609..5628)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6219..6238
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6523..6644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6560..6579)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6614..6633
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6693..6709)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6693..6709)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6706..6728)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6717..6733
                     /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(6741..6747,6748..6765,6766..6771))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6786..6807
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6924..6941)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7095..7680)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7175..7194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7851..8642,8643..8711))
                     /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     8474..8493
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8712..8816)
                     /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 ccacccagtg
     1801 tctgcctcag ggaaggagat ggattgggca cagatggcat gtcaccgatg tctggtgtat
     1861 ctgggggatt tgtcccgata tcagaatgaa ttagctggcg tagataccga gctgctagcc
     1921 gagagatttt actaccaagc cctgtcagta gctcctcaga ttggaatgcc cttcaatcag
     1981 ctgggcaccc tggcaggcag caagtactat aatgtggaag ccatgtattg ctacctgcgc
     2041 tgcatccagt cagaagtgtc ctttgaggga gcctatggga acctcaagcg gctgtatgac
     2101 aaggcagcca aaatgtacca ccaactgaag aagtgtgaga ctcggaaact gtctcctggc
     2161 aaaaagcgat gtaaagacat taaaaggttg ctagtgaact ttatgtatct gcaaagcctc
     2221 ctacagccca aaagcagctc cgtggactca gagctgacct cactttgcca gtcagtcctg
     2281 gaggacttca acctctgcct cttctacctg ccctcctcac ccaacctcag cctggccagt
     2341 gaggatgagg aggagtatga gagtggatat gctttcctcc cggaccttct catctttcaa
     2401 atggtcatca tctgccttat gtgtgtgcac agcttggaga gagcaggatc caagcagtac
     2461 agtgcagcca ttgccttcac cctggccctc ttttcccacc tcgtcaatca tgtcaacata
     2521 cggctgcagg ctgagctgga agagggcgag aatcccgtcc cggcattcca gagtgatggc
     2581 acagatgaac cagagtccaa ggaacctgtg gagaaagagg aggagccaga tcctgagcct
     2641 cctcctgtaa caccccaagt gggtgagggc agaaagagcc gtaagttctc tcgcctctcc
     2701 tgtctccgcc gtcgccgcca cccacccaaa gttggtgatg acagtgacct gagtgaaggc
     2761 tttgaatcgg actcaagcca tgactcagcc cgggccagtg agggctcaga cagtggctct
     2821 gacaagagtc ttgaaggtgg gggaacggcc tttgatgctg aaacagactc ggaaatgaat
     2881 agccaggagt cccgatcaga cttggaagat atggaggaag aggaggggac acggtcacca
     2941 accctggagc cccctcgggg cagatcagag gctcccgatt ccctcaatgg cccactgggc
     3001 cccagtgagg ctagcattgc cagcaatcta caagccatgt ccacccagat gttccagact
     3061 aagcgctgct tccgactggc ccccaccttt agcaacctgc tcctccagcc caccaccaac
     3121 cctcatacct cggccagcca caggccttgc gtcaatgggg atgtagacaa gccttcagag
     3181 ccagcctctg aggagggctc tgagtcggag gggagtgagt ccagtggacg ctcctgtcgg
     3241 aatgagcgca gcatccagga gaagcttcag gtcctgatgg ccgaaggtct gcttcctgct
     3301 gtgaaagtct tcctggactg gcttcggacc aaccccgacc tcatcatcgt gtgtgcacag
     3361 agctctcaaa gtctgtggaa ccgcctgtct gtgttgctga atctgttgcc tgctgctggt
     3421 gaactccagg agtctggcct ggccttgtgt cctgaggtcc aagatcttct tgaaggttgt
     3481 gaactgcctg acctcccctc tagccttctg ctcccagagg acatggctct tcgtaacctg
     3541 cccccgctcc gagctgccca cagacgcttt aactttgaca cggatcggcc cctgctcagc
     3601 accttagagg agtcagtggt gcgcatctgc tgcatccgca gctttggtca tttcatcgcc
     3661 cgcctgcaag gcagcatcct gcagttcaac ccagaggttg gcatcttcgt cagcattgcc
     3721 cagtctgagc aggagagcct gctgcagcag gcccaggcac agttccgaat ggcacaggag
     3781 gaagctcgtc ggaacaggct catgagagac atggctcagc tacgacttca gctcgaagtg
     3841 tctcagctgg agggcagcct gcagcagccc aaggcccagt cagccatgtc tccctacctc
     3901 gtccctgaca cccaggccct ctgccaccat ctccctgtca tccgccaact ggccaccagt
     3961 ggccgcttca ttgtcatcat cccaaggaca gtgatcgatg gcctggattt gctgaagaag
     4021 gaacacccag gggcccggga tgggattcgg tacctggagg cagagtttaa aaaaggaaac
     4081 aggtacattc gctgccagaa agaggtggga aagagctttg agcggcataa gctgaagagg
     4141 caggatgcag atgcctggac tctctataag atcctagaca gctgcaaaca gctgactctg
     4201 gcccaggggg caggtgagga ggatccgagt ggcatggtga ccatcatcac aggccttcca
     4261 ctggacaacc ccagcgtgct ttcaggcccc atgcaggcag ccctgcaggc cgctgcccac
     4321 gccagtgtgg acatcaagaa tgttctggac ttctacaagc agtggaagga aattggttga
     4381 gcggccgcct agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag
     4441 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     4501 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     4561 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     4621 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     4681 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     4741 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4801 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4861 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4921 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4981 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5041 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     5101 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     5161 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5221 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     5281 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     5341 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5401 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     5461 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     5521 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     5581 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     5641 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5701 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     5761 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5821 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5881 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5941 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     6001 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     6061 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     6121 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     6181 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     6241 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     6301 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     6361 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     6421 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     6481 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     6541 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6601 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     6661 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6721 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6781 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6841 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6901 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6961 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7021 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7081 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7141 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7201 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7261 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     7321 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7381 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7441 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7501 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     7561 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7621 gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7681 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7741 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7801 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7861 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7921 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7981 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     8041 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     8101 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     8161 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     8221 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8281 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8341 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8401 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8461 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8521 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8581 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8641 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8701 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8761 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8821 tttccccgaa aagtgccacc tgacgtc
//
