LOCUS       sequence-290211-        6733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg5-del267-809.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6733)
  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 6733)
  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-del267-809-V5-SBP-MBP_V
FEATURES             Location/Qualifiers
     source          1..6733
                     /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             2104..2145
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2167..2280
                     /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      3262..3271
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3521..3642
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3558..3577)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3612..3631
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3649..4104)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3786..3807)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3998..4017
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4131..4235
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4237..4594
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4237..4428
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4260..4280)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4445..4580
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4629..5423
                     /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             4629..5423
                     /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(4683..4702)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5293..5312
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5611..5630)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5655..5702
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6031..6619
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6520..6539
                     /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 gcacagttcc gaatggcaca ggaggaagct cgtcggaaca ggctcatgag agacatggct
     1501 cagctacgac ttcagctcga agtgtctcag ctggagggca gcctgcagca gcccaaggcc
     1561 cagtcagcca tgtctcccta cctcgtccct gacacccagg ccctctgcca ccatctccct
     1621 gtcatccgcc aactggccac cagtggccgc ttcattgtca tcatcccaag gacagtgatc
     1681 gatggcctgg atttgctgaa gaaggaacac ccaggggccc gggatgggat tcggtacctg
     1741 gaggcagagt ttaaaaaagg aaacaggtac attcgctgcc agaaagaggt gggaaagagc
     1801 tttgagcggc ataagctgaa gaggcaggat gcagatgcct ggactctcta taagatccta
     1861 gacagctgca aacagctgac tctggcccag ggggcaggtg aggaggatcc gagtggcatg
     1921 gtgaccatca tcacaggcct tccactggac aaccccagcg tgctttcagg ccccatgcag
     1981 gcagccctgc aggccgctgc ccacgccagt gtggacatca agaatgttct ggacttctac
     2041 aagcagtgga aggaaattgg tacgtcgacg gtaccgcggg cccgggatcc accggtccct
     2101 ccaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcccgg aagcggtagc
     2161 ggtaccatgg acgagaagac caccggctgg gggggcggcc acgtggtgga gggcctggcc
     2221 ggcgagctgg agcagctgcg ggccaggctg gagcaccacc ctcagggcca gcgggagccc
     2281 tccggcggct gcaagcccct cgacatgaaa atcgaagaag gtaaactggt aatctggatt
     2341 aacggcgata aaggctataa cggtctcgct gaagtcggta agaaattcga gaaagatacc
     2401 ggaattaaag tcaccgttga gcatccggat aaactggaag agaaattccc gcaggttgcg
     2461 gcaactggcg atggccctga cattatcttc tgggcacacg accgctttgg tggctacgct
     2521 caatctggcc tgttggctga aatcaccccg gacaaagcgt tccaggacaa gctgtatccg
     2581 tttacctggg atgccgtacg ttacaacggc aagctgattg cttacccgat cgctgttgaa
     2641 gcgttatcgc tgatttataa caaagatctg ctgccgaacc cgccaaaaac ctgggaagag
     2701 atcccggcgc tggataaaga actgaaagcg aaaggtaaga gcgcgctgat gttcaacctg
     2761 caagaaccgt acttcacctg gccgctgatt gctgctgacg ggggttatgc gttcaagtat
     2821 gaaaacggca agtacgacat taaagacgtg ggcgtggata acgctggcgc gaaagcgggt
     2881 ctgaccttcc tggttgacct gattaaaagc aaacacatga atgcagacac cgattactcc
     2941 atcgcagaag ctgcctttaa taaaggcgaa acagcgatga ccatcaacgg cccgtgggca
     3001 tggtccaaca tcgacaccag caaagtgaat tatggtgtaa cggtactgcc gaccttcaag
     3061 ggtcaaccat ccaaaccgtt cgttggcgtg ctgagcgcag gtattaacgc cgccagtccg
     3121 aacaaagagc tggcgaaaga gttcctcgaa aactatctgc tgactgatga aggtctggaa
     3181 gcggttaata aagacaaacc gctgggtgcc gtagcgctga agtcttacga ggaagagttg
     3241 gcgaaagatc cacgtattgc cgccaccatg gaaaacgccc agaaaggtga aatcatgccg
     3301 aacatcccgc agatgtccgc tttctggtat gccgtgcgta ctgcggtgat caacgccgcc
     3361 agcggtcgtc agactgtcga tgaagccctg aaagactaag cggccgcgac tctagatcat
     3421 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3481 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3541 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3601 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     3661 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3721 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3781 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3841 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3901 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3961 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     4021 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     4081 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4141 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4201 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     4261 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4321 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4381 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4441 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4501 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4561 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     4621 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4681 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4741 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4801 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4861 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4921 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4981 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     5041 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5101 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     5161 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5221 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5281 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5341 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5401 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5461 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5521 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5581 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5641 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5701 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5761 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5821 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     5881 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5941 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6001 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6061 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6121 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     6181 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6241 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6301 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6361 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6421 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6481 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6541 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6601 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6661 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6721 taccgccatg cat
//
