LOCUS       sequence-289920-        8551 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg7-K66ER163E-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8551)
  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 8551)
  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: pN1-HsSmg7-K66ER163E-siRNAres-V5-SBP-MBP_S
FEATURES             Location/Qualifiers
     source          1..8551
                     /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             3922..3963
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3985..4098
                     /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      5080..5089
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5339..5460
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5376..5395)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5430..5449
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5467..5922)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5604..5625)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5816..5835
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5949..6053
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6055..6412
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6055..6246
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6078..6098)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6263..6398
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6447..7241
                     /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             6447..7241
                     /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(6501..6520)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7111..7130
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7429..7448)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7473..7520
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7849..8437
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8338..8357
                     /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 atctcgagcc caacatgagc ctgcagagcg cgcagtacct ccggcaggca
      661 gaagtcctga aggctgacat gacagattct aagctgggtc cagctgaagt ctggacatcc
      721 aggcaggctc tgcaggacct gtaccagaaa atgctagtta ccgatttgga atacgcttta
      781 gacaagaaag tagaacagga tctctggaat cacgcctttg aaaatcagat cacaacactg
      841 caaggccagg caaagaatcg agcaaatccg aatcggagtg aagttcaggc aaacctttct
      901 ctgttcctag aggcagctag tggcttctat actcagttat tacaagaact gtgtacagta
      961 tttaatgtag atttaccatg ccgtgtgaag tcttcccaat tgggaattat cagcaataaa
     1021 cagacgcata ccagcgccat agtgaagcca cagtctagct cctgttccta tatctgccag
     1081 cactgcctcg tccaccttgg agacattgct gaatacagaa accagaccag ccaggcagag
     1141 tcctactata ggcatgcagc tcagcttgtc ccctccaatg gtcagcctta taatcagttg
     1201 gctatcttag cttcttccaa aggagaccat ctgaccacaa ttttctacta ctgcagaagc
     1261 attgctgtga agttcccttt cccagctgcc tccactaatc tgcaaaaagc actttctaaa
     1321 gcactggaaa gccgagatga ggtgaaaacc aagtggggtg tttctgactt catcaaggcc
     1381 tttattaaat tccacggtca tgtgtacctg agtaagagct tggaaaagtt gagccctctt
     1441 cgagagaaat tggaagaaca gtttaagagg ctgctattcc aaaaagcttt caactctcag
     1501 cagttagttc atgtcactgt cattaacctg tttcaacttc atcaccttcg tgactttagc
     1561 aatgaaaccg agcagcacac ttatagccaa gatgagcagc tatgttggac acagttgctg
     1621 gccctcttta tgtcttttct tggcatcctg tgcaagtgtc ctctacagaa tgagtctcag
     1681 gaggagtcct acaatgccta tcctcttcca gcagtcaagg tctccatgga ctggctaaga
     1741 ctcagaccca gggtctttca ggaggcagtg gtggatgaaa gacagtacat ttggccctgg
     1801 ttgatttccc ttctgaatag tttccatccc catgaagagg acctctcaag tattagtgcg
     1861 acaccacttc cagaggagtt tgaattacaa ggatttttgg cattgagacc ttctttcagg
     1921 aacttggatt tttccaaagg tcaccagggt attacagggg acaaagaagg ccagcaacga
     1981 cgaatacgac agcaacgctt gatctctata ggcaaatgga ttgctgataa tcagccaagg
     2041 ctgattcagt gtgaaaatga ggtagggaaa ttgttgttta tcacagaaat cccagaatta
     2101 atactggaag accccagtga agccaaagag aacctcattc tgcaggaaac atctgtgata
     2161 gagtcgctgg ctgcagatgg gagcccaggg ctaaaatcag tgctatctac aagccgaaat
     2221 ttaagcaaca actgtgacac aggagagaag ccagtggtta ccttcaaaga aaacattaag
     2281 acacgagaag tgaacagaga ccaaggaaga agttttcctc ccaaagaggt aaaatcccag
     2341 acagaactaa gaaagactcc agtgtctgaa gccagaaaaa cacctgtaac tcaaacccca
     2401 actcaagcaa gtaactccca gttcatcccc attcatcacc ctggagcctt ccctcctctt
     2461 cccagcaggc cagggtttcc gcccccaaca tatgttatcc ccccgcctgt ggcattttct
     2521 atgggctcag gttacacctt cccagctggt gtttctgtcc caggaacctt tcttcagcct
     2581 acagctcact ctccagcagg aaaccaggtg caagctggga aacagtccca cattccttac
     2641 agccagcaac ggccctctgg accagggcca atgaaccagg gacctcaaca atcacagcca
     2701 ccttcccagc aaccccttac atctttacca gctcagccta ctgctcaaag tactagtcaa
     2761 ctgcaggttc aagctctaac tcagcaacaa caatccccta caaaagctgt gccggctttg
     2821 gggaaaagcc cgcctcacca ctctggattc cagcagtatc aacaggcaga tgcctccaaa
     2881 cagctgtgga atccccctca ggttcaaggc ccattaggga aaattatgcc tgtgaaacag
     2941 ccctactacc ttcagaccca agaccccata aaactgtttg agccgtcatt gcaacctcct
     3001 gtaatgcagc agcagcctct agaaaaaaaa atgaagcctt ttcccatgga gccatataac
     3061 cataatccct cagaagtcaa ggtcccagaa ttctactggg attcttccta cagcatggct
     3121 gataacagat ctgtaatggc acagcaagca aacatagacc gcaggggcaa acggtcacca
     3181 ggaatcttcc gtccagagca ggatcctgta cccagaatgc cgtttgagaa atccttattg
     3241 gagaagccct cagagctcat gtcacattca tcctctttcc tgtccctcac cggattctct
     3301 ctcaatcagg aaagataccc aaataatagt atgttcaatg aggtatatgg gaaaaacctg
     3361 acatccagct ccaaagcaga actcagtccc tcaatggccc cccaggaaac atctctgtat
     3421 tccctttttg aagggactcc gtggtctcca tcacttcctg ccagttcaga tcattcaaca
     3481 ccagccagcc agtctcctca ttcctctaac ccaagcagcc tacccagctc tcctccaaca
     3541 cacaaccata attctgttcc attctccaat tttggaccca ttgggactcc agataacagg
     3601 gatagaagga ctgcagatcg gtggaaaact gataagccag ccatgggtgg gtttggcatt
     3661 gattatctct cagcaacgtc atcctctgag agcagttggc atcaggccag cactccgagt
     3721 ggcacctgga caggccatgg cccttccatg gaggattcct ctgctgtcct catggaaagc
     3781 ctaaagtcta tctggtccag ttccatgatg catcctggac cttctgctct ggagcagctg
     3841 ttaatgcagc agaagcagaa acagcaacgg ggacaaggca ccatgaaccc tccacactcc
     3901 cgggatccac cggtccctcc aggtaagcct atccctaacc ctctcctcgg tctcgattct
     3961 acgcccggaa gcggtagcgg taccatggac gagaagacca ccggctgggg gggcggccac
     4021 gtggtggagg gcctggccgg cgagctggag cagctgcggg ccaggctgga gcaccaccct
     4081 cagggccagc gggagccctc cggcggctgc aagcccctcg acatgaaaat cgaagaaggt
     4141 aaactggtaa tctggattaa cggcgataaa ggctataacg gtctcgctga agtcggtaag
     4201 aaattcgaga aagataccgg aattaaagtc accgttgagc atccggataa actggaagag
     4261 aaattcccgc aggttgcggc aactggcgat ggccctgaca ttatcttctg ggcacacgac
     4321 cgctttggtg gctacgctca atctggcctg ttggctgaaa tcaccccgga caaagcgttc
     4381 caggacaagc tgtatccgtt tacctgggat gccgtacgtt acaacggcaa gctgattgct
     4441 tacccgatcg ctgttgaagc gttatcgctg atttataaca aagatctgct gccgaacccg
     4501 ccaaaaacct gggaagagat cccggcgctg gataaagaac tgaaagcgaa aggtaagagc
     4561 gcgctgatgt tcaacctgca agaaccgtac ttcacctggc cgctgattgc tgctgacggg
     4621 ggttatgcgt tcaagtatga aaacggcaag tacgacatta aagacgtggg cgtggataac
     4681 gctggcgcga aagcgggtct gaccttcctg gttgacctga ttaaaagcaa acacatgaat
     4741 gcagacaccg attactccat cgcagaagct gcctttaata aaggcgaaac agcgatgacc
     4801 atcaacggcc cgtgggcatg gtccaacatc gacaccagca aagtgaatta tggtgtaacg
     4861 gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg ttggcgtgct gagcgcaggt
     4921 attaacgccg ccagtccgaa caaagagctg gcgaaagagt tcctcgaaaa ctatctgctg
     4981 actgatgaag gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt agcgctgaag
     5041 tcttacgagg aagagttggc gaaagatcca cgtattgccg ccaccatgga aaacgcccag
     5101 aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt tctggtatgc cgtgcgtact
     5161 gcggtgatca acgccgccag cggtcgtcag actgtcgatg aagccctgaa agactaagcg
     5221 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     5281 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     5341 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5401 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5461 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     5521 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     5581 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     5641 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     5701 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     5761 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     5821 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     5881 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     5941 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     6001 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     6061 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     6121 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     6181 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     6241 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     6301 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     6361 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     6421 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     6481 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     6541 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     6601 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     6661 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     6721 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     6781 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     6841 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     6901 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     6961 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     7021 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     7081 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     7141 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     7201 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     7261 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     7321 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     7381 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     7441 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     7501 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     7561 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     7621 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     7681 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7741 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7801 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7861 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7921 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7981 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     8041 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     8101 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     8161 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     8221 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     8281 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     8341 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     8401 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     8461 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     8521 ttctgtggat aaccgtatta ccgccatgca t
//
