LOCUS       sequence-290209-        7561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg5_267-1017.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7561)
  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 7561)
  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_267-1017-V5-SBP-MBP_V
FEATURES             Location/Qualifiers
     source          1..7561
                     /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             2932..2973
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2995..3108
                     /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      4090..4099
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4349..4470
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4386..4405)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4440..4459
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4477..4932)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4614..4635)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4826..4845
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4959..5063
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5065..5422
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5065..5256
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5088..5108)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5273..5408
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5457..6251
                     /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             5457..6251
                     /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(5511..5530)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6121..6140
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6439..6458)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6483..6530
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6859..7447
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7348..7367
                     /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 tgaagtgtga gactcggaaa
      661 ctgtctcctg gcaaaaagcg atgtaaagac attaaaaggt tgctagtgaa ctttatgtat
      721 ctgcaaagcc tcctacagcc caaaagcagc tccgtggact cagagctgac ctcactttgc
      781 cagtcagtcc tggaggactt caacctctgt ctcttctacc tgccctcctc acccaacctc
      841 agcctggcca gtgaggatga ggaggagtat gagagtggat atgctttcct cccggacctt
      901 ctcatctttc aaatggtcat catctgcctt atgtgtgtgc acagcttgga gagagcagga
      961 tccaagcagt acagtgcagc cattgccttc accctggccc tcttttccca cctcgtcaat
     1021 catgtcaaca tacggctgca ggctgagctg gaagagggcg agaatcccgt cccggcattc
     1081 cagagtgatg gcacagatga accagagtcc aaggaacctg tggagaaaga ggaggagcca
     1141 gatcctgagc ctcctcctgt aacaccccaa gtgggtgagg gcagaaagag ccgtaagttc
     1201 tctcgcctct cctgtctccg ccgtcgccgc cacccaccca aagttggtga tgacagtgac
     1261 ctgagtgaag gctttgaatc ggactcaagc catgactcag cccgggccag tgagggctca
     1321 gacagtggct ctgacaagag tcttgaaggt gggggaacgg cctttgatgc tgaaacagac
     1381 tcggaaatga atagccagga gtcccgatca gacttggaag atatggagga agaggagggg
     1441 acacggtcac caaccctgga gccccctcgg ggcagatcag aggctcccga ttccctcaat
     1501 ggcccactgg gccccagtga ggctagcatt gccagcaatc tacaagccat gtccacccag
     1561 atgttccaga ctaagcgctg cttccgactg gcccccacct ttagcaacct gctcctccag
     1621 cccaccacca accctcatac ctcggccagc cacaggcctt gcgtcaatgg ggatgtagac
     1681 aagccttcag agccagcctc tgaggagggc tctgagtcgg aggggagtga gtccagtgga
     1741 cgctcctgtc ggaatgagcg cagcatccag gagaagcttc aggtcctgat ggccgaaggt
     1801 ctgcttcctg ctgtgaaagt cttcctggac tggcttcgga ccaaccccga cctcatcatc
     1861 gtgtgtgcgc agagctctca aagtctgtgg aaccgcctgt ctgtgttgct gaatctgttg
     1921 cctgctgctg gtgaactcca ggagtctggc ctggccttgt gtcctgaggt ccaagatctt
     1981 cttgaaggtt gtgaactgcc tgacctcccc tctagccttc tgctcccaga ggacatggct
     2041 cttcgtaacc tgcccccgct ccgagctgcc cacagacgct ttaactttga cacggatcgg
     2101 cccctgctca gcaccttaga ggagtcagtg gtgcgcatct gctgcatccg cagctttggt
     2161 catttcatcg cccgcctgca aggcagcatc ctgcagttca acccagaggt tggcatcttc
     2221 gtcagcattg cccagtctga gcaggagagc ctgctgcagc aggcccaggc acagttccga
     2281 atggcacagg aggaagctcg tcggaacagg ctcatgagag acatggctca gctacgactt
     2341 cagctcgaag tgtctcagct ggagggcagc ctgcagcagc ccaaggccca gtcagccatg
     2401 tctccctacc tcgtccctga cacccaggcc ctctgccacc atctccctgt catccgccaa
     2461 ctggccacca gtggccgctt cattgtcatc atcccaagga cagtgatcga tggcctggat
     2521 ttgctgaaga aggaacaccc aggggcccgg gatgggattc ggtacctgga ggcagagttt
     2581 aaaaaaggaa acaggtacat tcgctgccag aaagaggtgg gaaagagctt tgagcggcat
     2641 aagctgaaga ggcaggatgc agatgcctgg actctctata agatcctaga cagctgcaaa
     2701 cagctgactc tggcccaggg ggcaggtgag gaggatccga gtggcatggt gaccatcatc
     2761 acaggccttc cactggacaa ccccagcgtg ctttcaggcc ccatgcaggc agccctgcag
     2821 gccgctgccc acgccagtgt ggacatcaag aatgttctgg acttctacaa gcagtggaag
     2881 gaaattggta cgtcgacggt accgcgggcc cgggatccac cggtccctcc aggtaagcct
     2941 atccctaacc ctctcctcgg tctcgattct acgcccggaa gcggtagcgg taccatggac
     3001 gagaagacca ccggctgggg gggcggccac gtggtggagg gcctggccgg cgagctggag
     3061 cagctgcggg ccaggctgga gcaccaccct cagggccagc gggagccctc cggcggctgc
     3121 aagcccctcg acatgaaaat cgaagaaggt aaactggtaa tctggattaa cggcgataaa
     3181 ggctataacg gtctcgctga agtcggtaag aaattcgaga aagataccgg aattaaagtc
     3241 accgttgagc atccggataa actggaagag aaattcccgc aggttgcggc aactggcgat
     3301 ggccctgaca ttatcttctg ggcacacgac cgctttggtg gctacgctca atctggcctg
     3361 ttggctgaaa tcaccccgga caaagcgttc caggacaagc tgtatccgtt tacctgggat
     3421 gccgtacgtt acaacggcaa gctgattgct tacccgatcg ctgttgaagc gttatcgctg
     3481 atttataaca aagatctgct gccgaacccg ccaaaaacct gggaagagat cccggcgctg
     3541 gataaagaac tgaaagcgaa aggtaagagc gcgctgatgt tcaacctgca agaaccgtac
     3601 ttcacctggc cgctgattgc tgctgacggg ggttatgcgt tcaagtatga aaacggcaag
     3661 tacgacatta aagacgtggg cgtggataac gctggcgcga aagcgggtct gaccttcctg
     3721 gttgacctga ttaaaagcaa acacatgaat gcagacaccg attactccat cgcagaagct
     3781 gcctttaata aaggcgaaac agcgatgacc atcaacggcc cgtgggcatg gtccaacatc
     3841 gacaccagca aagtgaatta tggtgtaacg gtactgccga ccttcaaggg tcaaccatcc
     3901 aaaccgttcg ttggcgtgct gagcgcaggt attaacgccg ccagtccgaa caaagagctg
     3961 gcgaaagagt tcctcgaaaa ctatctgctg actgatgaag gtctggaagc ggttaataaa
     4021 gacaaaccgc tgggtgccgt agcgctgaag tcttacgagg aagagttggc gaaagatcca
     4081 cgtattgccg ccaccatgga aaacgcccag aaaggtgaaa tcatgccgaa catcccgcag
     4141 atgtccgctt tctggtatgc cgtgcgtact gcggtgatca acgccgccag cggtcgtcag
     4201 actgtcgatg aagccctgaa agactaagcg gccgcgactc tagatcataa tcagccatac
     4261 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     4321 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     4381 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4441 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     4501 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4561 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4621 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4681 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4741 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4801 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4861 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4921 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4981 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5041 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     5101 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5161 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     5221 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     5281 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     5341 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     5401 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     5461 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5521 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5581 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     5641 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5701 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5761 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5821 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5881 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5941 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     6001 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     6061 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     6121 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     6181 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6241 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     6301 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6361 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6421 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     6481 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     6541 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     6601 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     6661 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     6721 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6781 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6841 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6901 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6961 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     7021 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7081 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7141 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7201 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7261 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7321 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7381 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7441 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7501 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     7561 t
//
