LOCUS       sequence-289914-        7057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of CeSmg7-T83E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7057)
  AUTHORS   Jonas S, Weichenrieder O, Izaurralde E
  TITLE     An unusual arrangement of two 14-3-3-like domains in the SMG5-SMG7 
            heterodimer is required for efficient nonsense-mediated mRNA decay.
  JOURNAL   Genes Dev. 2013 Jan 15;27(2):211-25. doi: 10.1101/gad.206672.112.
  PUBMED    23348841
REFERENCE   2  (bases 1 to 7057)
  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.206672"; journalName: "Genes Dev"; date: "2013-01-15- 
            15"; volume: "27"; issue: "2"; pages: "211-25"
COMMENT     Sequence Label: pCIneo-V5-SBP-CeSmg7-T83E_S
FEATURES             Location/Qualifiers
     source          1..7057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1098..1107
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1107..1148
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1170..1283
                     /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"
     polyA_signal    2761..2882
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2772..2791)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2826..2845
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3068..3523
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3155..3174)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3365..3386
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3640..3997
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3640..3831
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3663..3683)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3848..3983
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4048..4842
                     /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(4102..4121)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4712..4731
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4906..4954
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4995..5014)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5114..5136
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5174..5192)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5260..5364
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5365..5433,5434..6225)
                     /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     complement(5583..5602)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6396..6984
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6885..6904
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc aattgcggcc accatgggta agcctatccc taaccctctc ctcggtctcg
     1141 attctacgcc cggaagcggt agcggtacca tggacgagaa gaccaccggc tgggggggcg
     1201 gccacgtggt ggagggcctg gccggcgagc tggagcagct gcgggccagg ctggagcacc
     1261 accctcaggg ccagcgggag ccctccggcg gctgcaagcc cctcgagaat tcacgcgtgg
     1321 tacctctaga gtcgaccatg tcggacgaat gggaacagct tactgtagag ttgagaaaaa
     1381 ttccacgtgg caccgaagct gcaccacagt atttgagaca tttaatgaaa atgtttgtcg
     1441 ctgacttcga aactgctgta tccaaaagat tcgatgtaaa attctggaat aaactgaaat
     1501 caatgatgga tgaaataaca aaagcaatgg aaaatgatcg acttgttaat cataatgttc
     1561 aaaatcttgc aattggattt ttggaggatt tatcacttct tgttcattat cattatgaaa
     1621 ttccaaatta tggaaacgat atctcaaaac aattaacatg gacaccagat gtatttctaa
     1681 acagaaaacc tatcaagtcg aaaaagaata gtcgagtgtt catggcttat gttctgttaa
     1741 gaatgggaga tttaatgaga tataaggaga actacccaaa agctcaggaa tattacgagc
     1801 aatcatgccg cattaatcca gctgatggag ctgtctggaa tcagctagga ttgatatctt
     1861 cacttggtgc aaaaaacctg gaaagtgtct attttcatac acgagctctt catgccacta
     1921 tggaattccc aactgcttct ggtggcttaa cgaatatttt caaaaatttt gcaaatcgag
     1981 atatttcaag accaatgcct attaaagatc tatatttatc ctgcctagga agaattcatt
     2041 tccttctaga aatcgaagat tcttctgtgc acctacaaaa aattggagaa gaagctgcaa
     2101 cttcaaaaga aatgattgtt cctctaatgt ctgtatataa acatctcgag gacggaactg
     2161 aacttgaaca acgagccgta gagtatgtca aaactatttg gtgtacggcc taccgaagtc
     2221 tcctgaaaac tttggatgat tataaggagg aatcaaaaaa actggcagat gttccacatc
     2281 ttcttcatat tctggcactt ttactctgtg cacctaaact ccttcgcggt attgaagacc
     2341 aaactgaaga tgaagtaaca tcaatttgcg aatggcttct atgcgcaaat tgtgatgaga
     2401 aaatcaagga ttctgatgct tttggatatt ttcattgtct tcaaagaatt caatatccac
     2461 tcactagaac tcaattggct cagaaattag ttgaaataga ggatgaagat gaatctaaag
     2521 attccgatca atcaacaata ccagaagaag atcagaaaag cctttctaca gacacttcaa
     2581 gaattcaaaa cgaggattcg gaacctgaat ctgaatctga atcttcggat gaggaagttt
     2641 tgcaacgtgg ccgtcggcgt ggaagaaatc cgaatctaga cgaatcagtc gaaacggatg
     2701 aagatgaatt ttaagcggcc gcttcccttt agtgagggtt aatgcttcga gcagacatga
     2761 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2821 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2881 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     2941 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatccg ataaggatcg atccgggctg
     3001 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     3061 cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3121 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3181 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3241 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3301 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3361 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3421 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3481 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc
     3541 tccttacgca tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc
     3601 tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct
     3661 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3721 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3781 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3841 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3901 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3961 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttgattctt ctgacacaac
     4021 agtctcgaac ttaaggctag agccaccatg attgaacaag atggattgca cgcaggttct
     4081 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4141 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4201 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4261 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4321 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4381 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4441 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4501 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4561 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4621 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4681 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4741 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4801 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4861 aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa aatatcttta
     4921 ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag gatccgcgta
     4981 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     5041 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     5101 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     5161 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5221 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5281 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5341 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5401 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5461 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5521 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5581 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5641 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5701 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5761 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5821 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5881 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5941 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6001 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6061 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6121 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6181 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6241 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6301 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6361 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6421 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6481 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6541 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6601 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6661 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6721 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6781 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6841 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6901 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6961 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7021 ttttgctggc cttttgctca catggctcga cagatct
//
