LOCUS       sequence-289924-        7277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsUpf1_115-1129-S1107A-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7277)
  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 7277)
  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: pT7-V5-SBP-C1-HsUpf1_115-1129-S1107A-siRNAres_S
FEATURES             Location/Qualifiers
     source          1..7277
                     /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"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             668..709
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             731..844
                     /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    4065..4186
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4102..4121)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4156..4175
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4193..4648)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4330..4351)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4542..4561
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4675..4779
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4781..5138
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4781..4972
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4804..4824)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4989..5124
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5173..5967
                     /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             5173..5967
                     /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(5227..5246)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5837..5856
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6155..6174)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6199..6246
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6575..7163
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7064..7083
                     /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 gcagagctct ctggctaact agagaaccca ctgcttactg
      601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      661 caccatgggt aagcctatcc ctaaccctct cctcggtctc gattctacgc ccggaagcgg
      721 tagcggtacg atggacgaga agaccaccgg ctgggggggc ggccacgtgg tggagggcct
      781 ggccggcgag ctggagcagc tgcgggccag gctggagcac caccctcagg gccagcggga
      841 gcccagatct cgagctacga aggacctccc catacacgcc tgcagttact gtggaataca
      901 cgatcctgcc tgcgtggttt actgtaatac cagcaagaag tggttctgca acggacgtgg
      961 aaatacttct ggcagccaca ttgtaaatca ccttgtgagg gcaaaatgca aaggggtgac
     1021 cctgcacaag gacgggcccc tgggggagac agtcctggag tgctacaact gcggctgtcg
     1081 caacgtcttc ctcctcggct tcatcccggc caaagctgac tcagtggtgg tgctgctgtg
     1141 caggcagccc tgtgccagcc agagcagcct caaggacatc aactgggaca gctcgcagtg
     1201 gcagccgctg atccaggacc gctgcttcct gtcctggctg gtcaagatcc cctccgagca
     1261 ggagcagctg cgggcacgcc agatcacggc acagcagatc aacaagctgg aggagctgtg
     1321 gaaggaaaac ccttctgcca cgctggagga cctggagagg ccgggggtgg acgaggagcc
     1381 gcagcatgtc ctcctgcggt acgaggacgc ctaccagtac cagaacatat tcgggcccct
     1441 ggtcaagctg gaggccgact acgacaagaa gctgaaggag tcccagactc aagataacat
     1501 cactgtcagg tgggacctgg gccttaacaa gaagagaatc gcctacttca ctttgcccaa
     1561 gactgactct ggtaatgagg atttagtcat aatttggtta agagacatgc ggctcatgca
     1621 gggggatgag atatgcctgc ggtacaaagg ggaccttgcg cccctgtgga aagggatcgg
     1681 ccacgtcatc aaggtccctg ataattatgg cgatgagatc gccattgagc tgcggagcag
     1741 cgtgggtgca cctgtggagg tgactcacaa cttccaggtg gattttgtgt ggaagtcgac
     1801 ctcctttgac aggatgcaga gcgcattgaa aacgtttggc gtggatgaga cctcggtgtc
     1861 tggctacatc taccacaagc tgttgggcca cgaggtggag gacgtaatca tcaagtgcca
     1921 gctgcccaag cgcttcacgg cgcagggcct ccccgacctc aaccactccc aggtttatgc
     1981 cgtgaagact gtgctgcaaa gaccactgag cctgatccag ggcccgccag gcacggggaa
     2041 gacggtgacg tcggccacca tcgtctacca cctggcccgg caaggcaacg ggccggtgct
     2101 ggtgtgtgct ccgagcaaca tcgccgtgga ccagctaacg gagaagatcc accagacggg
     2161 gctaaaggtc gtgcgcctct gcgccaagag ccgtgaggcc atcgactccc cggtgtcttt
     2221 tctggccctg cacaaccaga tcaggaacat ggacagcatg cctgagctgc agaagctgca
     2281 gcagctgaaa gacgagactg gggagctgtc gtctgccgac gagaagcggt accgggcctt
     2341 gaagcgcacc gcagagagag agctgctgat gaacgcagat gtcatctgct gcacatgtgt
     2401 gggcgccggt gacccgaggc tggccaaaat gcaatttagg agtattttaa tcgacgaaag
     2461 cacccaggcc accgagccgg agtgcatggt tcccgtggtc ctcggggcca agcagctgat
     2521 ccttgtaggc gaccactgcc agctgggccc agtggtgatg tgcaagaagg cggccaaggc
     2581 cgggctgtca cagtcgctct tcgagcgcct ggtggtgctg ggcatccggc ccatccgcct
     2641 gcaggtccag taccggatgc accctgcact cagcgccttc ccatccaaca tcttctacga
     2701 gggctccctc cagaatggtg tcactgcagc ggatcgtgtg aagaagggat ttgacttcca
     2761 gtggccccaa cccgataaac cgatgttctt ctacgtgacc cagggccaag aggagattgc
     2821 cagctcgggc acctcctacc tgaacaggac cgaggctgcg aacgtggaga agatcaccac
     2881 gaagttgctg aaggcaggcg ccaagccgga ccagattggc atcatcacgc cctacgaggg
     2941 ccagcgctcc tacctggtgc agtacatgca gttcagcggc tccctgcaca ccaagctcta
     3001 ccaggaggtg gagatcgcca gtgtggacgc ctttcaggga cgcgagaagg acttcatcat
     3061 cctgtcctgt gtgcgggcca acgagcacca aggcattggc tttttaaatg accccaggcg
     3121 tctgaacgtg gccctgacca gagcaaggta tggcgtcatc attgtgggca acccgaaggc
     3181 actatcaaag cagccgctct ggaaccacct gctgaactac tataaggagc agaaggtgct
     3241 ggtggagggg ccgctcaaca acctgcgtga gagcctcatg cagttcagca agccacggaa
     3301 gctggtcaac actatcaacc cgggagcccg cttcatgacc acagccatgt atgatgcccg
     3361 ggaggccatc atcccaggct ccgtctatga tcggagcagc cagggccggc cttccagcat
     3421 gtacttccag acccatgacc agattggcat gatcagtgcc ggccctagcc acgtggctgc
     3481 catgaacatt cccatcccct tcaacctggt catgccaccc atgccaccgc ctggctattt
     3541 tggacaagcc aacgggcctg ctgcagggcg aggcaccccg aaaggcaaga ctggtcgtgg
     3601 gggacgccag aagaaccgct ttgggcttcc tggacccagc cagactaacc tccccaacag
     3661 ccaagccagc caggatgtgg cgtcacagcc cttctctcag ggcgccctga cgcagggcta
     3721 catctccatg agccagcctt cccagatgag ccagcccggc ctctcccagc cggagctgtc
     3781 ccaggacagt taccttggtg acgagtttaa atcacaaatc gacgtggcgc tcgcacagga
     3841 ctccacgtac cagggagagc gggcttacca gcatggcggg gtgatggggc tgtcccagta
     3901 ttaagaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga
     3961 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     4021 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     4081 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4141 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     4201 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4261 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     4321 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4381 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     4441 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4501 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4561 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4621 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4681 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4741 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4801 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4861 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4921 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4981 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5041 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     5101 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     5161 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5221 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5281 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5341 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5401 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5461 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5521 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5581 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5641 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5701 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5761 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5821 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5881 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5941 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     6001 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     6061 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     6121 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     6181 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     6241 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     6301 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     6361 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     6421 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6481 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6541 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6601 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6661 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6721 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6781 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6841 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6901 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6961 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     7021 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     7081 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7141 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7201 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     7261 gtattaccgc catgcat
//
