LOCUS       sequence-290212-        8248 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian Expression of HsSmg5-del810-846.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8248)
  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 8248)
  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"; 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-del810-846-V5-SBP-MBP_V
FEATURES             Location/Qualifiers
     source          1..8248
                     /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             3619..3660
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3682..3795
                     /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      4777..4786
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5036..5157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5073..5092)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5127..5146
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5164..5619)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5301..5322)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5513..5532
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5646..5750
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5752..6109
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5752..5943
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5775..5795)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5960..6095
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6144..6938
                     /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             6144..6938
                     /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(6198..6217)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6808..6827
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7126..7145)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7170..7217
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7546..8134
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8035..8054
                     /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 gcccagccca aggcccagtc agccatgtct ccctacctcg tccctgacac ccaggccctc
     3121 tgccaccatc tccctgtcat ccgccaactg gccaccagtg gccgcttcat tgtcatcatc
     3181 ccaaggacag tgatcgatgg cctggatttg ctgaagaagg aacacccagg ggcccgggat
     3241 gggattcggt acctggaggc agagtttaaa aaaggaaaca ggtacattcg ctgccagaaa
     3301 gaggtgggaa agagctttga gcggcataag ctgaagaggc aggatgcaga tgcctggact
     3361 ctctataaga tcctagacag ctgcaaacag ctgactctgg cccagggggc aggtgaggag
     3421 gatccgagtg gcatggtgac catcatcaca ggccttccac tggacaaccc cagcgtgctt
     3481 tcaggcccca tgcaggcagc cctgcaggcc gctgcccacg ccagtgtgga catcaagaat
     3541 gttctggact tctacaagca gtggaaggaa attggtacgt cgacggtacc gcgggcccgg
     3601 gatccaccgg tccctccagg taagcctatc cctaaccctc tcctcggtct cgattctacg
     3661 cccggaagcg gtagcggtac catggacgag aagaccaccg gctggggggg cggccacgtg
     3721 gtggagggcc tggccggcga gctggagcag ctgcgggcca ggctggagca ccaccctcag
     3781 ggccagcggg agccctccgg cggctgcaag cccctcgaca tgaaaatcga agaaggtaaa
     3841 ctggtaatct ggattaacgg cgataaaggc tataacggtc tcgctgaagt cggtaagaaa
     3901 ttcgagaaag ataccggaat taaagtcacc gttgagcatc cggataaact ggaagagaaa
     3961 ttcccgcagg ttgcggcaac tggcgatggc cctgacatta tcttctgggc acacgaccgc
     4021 tttggtggct acgctcaatc tggcctgttg gctgaaatca ccccggacaa agcgttccag
     4081 gacaagctgt atccgtttac ctgggatgcc gtacgttaca acggcaagct gattgcttac
     4141 ccgatcgctg ttgaagcgtt atcgctgatt tataacaaag atctgctgcc gaacccgcca
     4201 aaaacctggg aagagatccc ggcgctggat aaagaactga aagcgaaagg taagagcgcg
     4261 ctgatgttca acctgcaaga accgtacttc acctggccgc tgattgctgc tgacgggggt
     4321 tatgcgttca agtatgaaaa cggcaagtac gacattaaag acgtgggcgt ggataacgct
     4381 ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaagcaaaca catgaatgca
     4441 gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc gatgaccatc
     4501 aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg tgtaacggta
     4561 ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag cgcaggtatt
     4621 aacgccgcca gtccgaacaa agagctggcg aaagagttcc tcgaaaacta tctgctgact
     4681 gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc gctgaagtct
     4741 tacgaggaag agttggcgaa agatccacgt attgccgcca ccatggaaaa cgcccagaaa
     4801 ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt gcgtactgcg
     4861 gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga ctaagcggcc
     4921 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     4981 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     5041 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5101 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     5161 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     5221 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     5281 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     5341 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     5401 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     5461 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     5521 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     5581 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     5641 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5701 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     5761 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     5821 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     5881 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     5941 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     6001 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     6061 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     6121 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     6181 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     6241 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     6301 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     6361 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     6421 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     6481 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     6541 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     6601 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     6661 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     6721 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     6781 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     6841 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     6901 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     6961 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     7021 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     7081 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     7141 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     7201 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     7261 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     7321 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     7381 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     7441 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7501 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     7561 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     7621 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     7681 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     7741 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     7801 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     7861 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     7921 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     7981 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     8041 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     8101 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     8161 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     8221 tgtggataac cgtattaccg ccatgcat
//
