LOCUS       sequence-289015-        6982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg6_1-576.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6982)
  AUTHORS   Kashima I, Jonas S, Jayachandran U, Buchwald G, Conti E, Lupas AN, 
            Izaurralde E
  TITLE     SMG6 interacts with the exon junction complex via two conserved 
            EJC-binding motifs (EBMs) required for nonsense-mediated mRNA decay.
  JOURNAL   Genes Dev. 2010 Nov 1. 24(21):2440-50.
  PUBMED    20930030
REFERENCE   2  (bases 1 to 6982)
  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"; journalName: "Genes Dev. 
            2010 Nov 1. 24(21):2440-50."
COMMENT     Sequence Label: pN1-HsSmg6_1-576-V5-SBP-MBP_I
FEATURES             Location/Qualifiers
     source          1..6982
                     /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"
     regulatory      604..613
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2353..2394
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2416..2529
                     /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      3511..3520
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3770..3891
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3807..3826)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3861..3880
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3898..4353)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4035..4056)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4247..4266
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4380..4484
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4486..4843
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4486..4677
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4509..4529)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4694..4829
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4878..5672
                     /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             4878..5672
                     /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(4932..4951)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5542..5561
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5860..5879)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5904..5951
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6280..6868
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 gctagcaatt
      601 gcggccacca tggcggaagg gctggagcgt gtgcggatct ccgcgtcgga gctgcgcggg
      661 atcctggcta ctctggcccc gcaggccggg agcagagaaa acatgaagga attaaaggag
      721 gccaggccgc gcaaagataa caggcgtcca gatctggaaa tctataagcc tggcctttct
      781 cggctaagga acaagcccaa aatcaaggaa ccccctggga gtgaggaatt caaagatgaa
      841 attgttaatg accgagattg ctctgctgtt gaaaatggta cacagcccgt taaagatgtc
      901 tgcaaggaac tgaacaacca agagcagaat ggtcctatag acccagaaaa taatcgggga
      961 caagaatcct ttcctaggac tgctggacaa gaggatcgta gtctaaaaat tatcaaaaga
     1021 acaaagaaac ccgacctgca gatctatcag cctggacgac gtttgcagac tgttagcaaa
     1081 gaatccgcca gtcgggtgga ggaggaagaa gtcctcaacc aggtagaaca actgagagta
     1141 gaggaagatg agtgtagggg aaatgttgcg aaggaggaag ttgcgaataa accagacagg
     1201 gccgagatag aaaagagccc aggtggtggg agagtagggg ctgcaaaagg agaaaaagga
     1261 aagaggatgg gaaaagggga gggggtgagg gaaacccacg acgacccggc ccgcgggagg
     1321 ccgggctccg caaagcgcta ctcccgctca gacaaacgaa ggaatcgcta ccgcacgcgc
     1381 agcaccagct cagctggcag caacaacagc gctgagggag ctggcctgac ggataatgga
     1441 tgtcgccgcc gccgacagga taggaccaag gagaggccac cactgaagaa gcaagtgtct
     1501 gtgtcctcaa ccgattcctt agacgaggac agaattgatg agcctgatgg attaggaccc
     1561 aggagaagtt cagaaaggaa gagacattta gaaagaaact ggtctggccg tggggagggt
     1621 gagcagaaaa ccagtgctaa agaatatcga ggcactcttc gtgtcacttt cgatgcagaa
     1681 gccatgaaca aagagtctcc catggtgagg tcagccaggg atgatatgga tagaggaaag
     1741 cctgacaaag gcttgagcag tgggggcaaa ggctctgaga agcaggagtc caaaaacccg
     1801 aaacaagaac ttcggggtcg tggtcgtggc attctgattt tgcctgccca taccacccta
     1861 tctgtcaatt cagcaggttc tccagagtcc gcgcctttgg gacctcggct tttgtttgga
     1921 tctggtagta agggatctcg gagttggggc cgtggaggca ccacacgccg attgtgggac
     1981 ccaaacaatc ctgatcagaa acctgctcta aagactcaga cgccccagct acatttcttg
     2041 gacactgatg atgaagtcag ccctacatct tggggtgact cacgccaggc tcaggcatct
     2101 tactataagt ttcaaaactc tgacaacccc tattattacc cccggacacc aggccctgcc
     2161 tcccagtatc cctatacggg ctataaccct ctacagtacc cagtgggccc tacgaatggt
     2221 gtgtacccag ggccttacta cccaggctac ccgactccgt caggacagta tgtgtgtagc
     2281 cctctaccta ccagcaccat gagtcccgag gaggtagagc agcacatgag gaacctgcca
     2341 ccggtccctc caggtaagcc tatccctaac cctctcctcg gtctcgattc tacgcccgga
     2401 agcggtagcg gtaccatgga cgagaagacc accggctggg ggggcggcca cgtggtggag
     2461 ggcctggccg gcgagctgga gcagctgcgg gccaggctgg agcaccaccc tcagggccag
     2521 cgggagccct ccggcggctg caagcccctc gagatgaaaa tcgaagaagg taaactggta
     2581 atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa gaaattcgag
     2641 aaagataccg gaattaaagt caccgttgag catccggata aactggaaga gaaattcccg
     2701 caggttgcgg caactggcga tggccctgac attatcttct gggcacacga ccgctttggt
     2761 ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt ccaggacaag
     2821 ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc ttacccgatc
     2881 gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc gccaaaaacc
     2941 tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag cgcgctgatg
     3001 ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg gggttatgcg
     3061 ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa cgctggcgcg
     3121 aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa tgcagacacc
     3181 gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac catcaacggc
     3241 ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac ggtactgccg
     3301 accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg tattaacgcc
     3361 gccagtccga acaaagagct ggcgaaagag ttcctcgaaa actatctgct gactgatgaa
     3421 ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa gtcttacgag
     3481 gaagagttgg cgaaagatcc acgtattgcc gccaccatgg aaaacgccca gaaaggtgaa
     3541 atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac tgcggtgatc
     3601 aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagactaagc ggccgcgact
     3661 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     3721 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3781 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3841 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     3901 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     3961 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     4021 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     4081 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     4141 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     4201 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     4261 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     4321 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     4381 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4441 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     4501 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4561 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4621 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4681 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4741 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4801 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     4861 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4921 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4981 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     5041 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     5101 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     5161 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5221 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5281 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5341 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5401 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5461 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5521 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5581 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5641 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5701 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5761 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5821 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     5881 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     5941 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     6001 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     6061 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     6121 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6181 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6241 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6301 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6361 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6421 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6481 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6541 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6601 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6661 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6721 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6781 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6841 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6901 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6961 taaccgtatt accgccatgc at
//
