LOCUS       sequence-289004-        8390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg6_1-1238-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8390)
  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 8390)
  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: pEYFP-N1-HsSmg6_1-1238-siRNAres_I
FEATURES             Location/Qualifiers
     source          1..8390
                     /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      4330..4339
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4336..4338,4339..4341,4342..5055)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4381..4402)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4642..4661)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4989..5010
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5178..5299
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5215..5234)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5269..5288
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5306..5761)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5443..5464)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5655..5674
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5788..5892
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5894..6251
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5894..6085
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5917..5937)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6102..6237
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6286..7080
                     /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             6286..7080
                     /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(6340..6359)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6950..6969
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7268..7287)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7312..7359
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7688..8276
                     /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 gctagccaac
      601 atggcggaag ggctggagcg tgtgcggatc tccgcgtcgg agctgcgcgg gatcctggct
      661 actctggccc cgcaggccgg gagcagagaa aacatgaagg aattaaagga ggccaggccg
      721 cgcaaagata acaggcgtcc agatctggaa atctataagc ctggcctttc tcggctaagg
      781 aacaagccca aaatcaagga accccctggg agtgaggaat tcaaagatga aattgttaat
      841 gaccgagatt gctctgctgt tgaaaatggt acacagcccg ttaaagatgt ctgcaaggaa
      901 ctgaacaacc aagagcagaa tggtcctata gacccagaaa ataatcgggg acaagaatcc
      961 tttcctagga ctgctggaca agaggatcgt agtctaaaaa ttatcaaaag aacaaagaaa
     1021 cccgacctgc agatctatca gcctggacga cgtttgcaga ctgttagcaa agaatccgcc
     1081 agtcgggtgg aggaggaaga agtcctcaac caggtagaac aactgagagt agaggaagat
     1141 gagtgtaggg gaaatgttgc gaaggaggaa gttgcgaata aaccagacag ggccgagata
     1201 gaaaagagcc caggtggtgg gagagtaggg gctgcaaaag gagaaaaagg aaagaggatg
     1261 ggaaaagggg agggggtgag ggaaacccac gacgacccgg cccgcgggag gccgggctcc
     1321 gcaaagcgct actcccgctc agacaaacga aggaatcgct accgcacgcg cagcaccagc
     1381 tcagctggca gcaacaacag cgctgaggga gctggcctga cggataatgg atgtcgccgc
     1441 cgccgacagg ataggaccaa ggagaggcca ccactgaaga agcaagtgtc tgtgtcctca
     1501 accgattcct tagacgagga cagaattgat gagcctgatg gattaggacc caggagaagt
     1561 tcagaaagga agagacattt agaaagaaac tggtctggcc gtggggaggg tgagcagaaa
     1621 accagtgcta aagaatatcg aggcactctt cgtgtcactt tcgatgcaga agccatgaac
     1681 aaagagtctc ccatggtgag gtcagccagg gatgatatgg atagaggaaa gcctgacaaa
     1741 ggcttgagca gtgggggcaa aggctctgag aagcaggagt ccaaaaaccc gaaacaagaa
     1801 cttcggggtc gtggtcgtgg cattctgatt ttgcctgccc ataccaccct atctgtcaat
     1861 tcagcaggtt ctccagagtc cgcgcctttg ggacctcggc ttttgtttgg atctggtagt
     1921 aagggatctc ggagttgggg ccgtggaggc accacacgcc gattgtggga cccaaacaat
     1981 cctgatcaga aacctgctct aaagactcag acgccccagc tacatttctt ggacactgat
     2041 gatgaagtca gccctacatc ttggggtgac tcacgccagg ctcaggcatc ttactataag
     2101 tttcaaaact ctgacaaccc ctattattac ccccggacac caggccctgc ctcccagtat
     2161 ccctatacgg gctataaccc tctacagtac ccagtgggcc ctacgaatgg tgtgtaccca
     2221 gggccttact acccaggcta cccgactccg tcaggacagt atgtgtgtag ccctctacct
     2281 accagcacca tgagtcccga ggaggtagag cagcacatga ggaacctgca gcaacaggag
     2341 ctgcacaggc ttctccgggt ggctgacaac caggaactgc agctcagcaa cctgctctcc
     2401 agggaccgca tcagtccgga gggcctggag aagatggcgc aactcagagc tgaactgctg
     2461 cagctatatg agcgctgtat tctattagat attgagttct ctgataatca gaatgtggat
     2521 cagatcctgt ggaagaatgc tttctatcag gtgattgaga agttcaggca acttgtcaag
     2581 gatccgaatg ttgagaaccc agaacagatt cggaacagac ttttggagct cttggatgag
     2641 ggtagtgact tctttgatag tttgcttcag aagctgcagg ttacttacaa gttcaaactg
     2701 gaagactaca tggatggtct tgccattcgc agcaagccat tacgcaagac agtaaaatat
     2761 gccttgatca gtgcccagcg atgcatgata tgccaaggag atattgctag gtaccgggag
     2821 caggcaagcg acacggcaaa ctatgggaaa gcacgcagtt ggtacctgaa ggcccagcac
     2881 attgctccca agaatgggcg cccctataac cagttggctt tgctggcagt gtatacgagg
     2941 aggaagcttg acgctgtcta ttactatatg cgcagtttag ctgccagcaa ccctatcctg
     3001 actgccaagg agagtctcat gagcttgttt gaagagacca agcggaaggc agaacagatg
     3061 gaaaagaagc aacatgagga atttgacctg agccctgacc agtggcggaa aggaaagaag
     3121 tctactttcc ggcatgttgg agatgacacc actcgcctgg agatctggat tcatccatcc
     3181 catccacggt cttcccaggg cactgagtct gggaaggatt ctgagcaaga gaatgggctg
     3241 ggcagcctga gtcccagtga tctgaacaaa aggttcatcc tcagttttct ccatgcccat
     3301 gggaagctgt ttacccggat tgggatggag acattccctg cagtggctga gaaggtcctc
     3361 aaggagttcc aggtgttact gcagcacagc ccctctccca ttggaagtac ccgcatgctg
     3421 cagcttatga ccatcaatat gtttgcagta cacaactccc agctgaaaga ctgcttctcg
     3481 gaggagtgcc gctctgtgat ccaggaacaa gccgcagctc tgggcttggc catgttttct
     3541 ctactggtcc gccgctgcac ctgcttactt aaggagtccg ccaaagctca gctgtcctct
     3601 cctgaggacc aggatgacca agacgacatc aaggtgtctt cctttgtccc ggacctgaag
     3661 gagctgctcc ccagtgtcaa agtctggtca gattggatgc tcggctaccc ggacacctgg
     3721 aatcctcctc ccacatccct ggatctgccc tcgcatgttg ctgtggatgt atggtcgacg
     3781 ctggctgatt tctgtaacat actgactgca gtgaatcagt ctgaggtgcc actgtacaag
     3841 gacccggatg atgacctcac ccttcttatc ctggaagagg atcggcttct ctcgggcttt
     3901 gtccccttgc tggctgcccc tcaggacccc tgctacgtgg agaaaacctc ggataaggtt
     3961 attgcagctg actgcaaaag ggtcacagtg ctgaagtatt ttctggaagc cctttgtgga
     4021 caagaagagc ctctgctggc attcaagggt ggaaagtatg tgtcagtggc acccgtccca
     4081 gacaccatgg gaaaggaaat gggaagccaa gagggaacac gactggaaga tgaggaggag
     4141 gatgtggtga ttgaagactt tgaggaagat tcagaggctg aaggcagcgg aggcgaggat
     4201 gacatcaggg agcttcgggc caagaagctg gctctggcca ggaagatagc tgagcagcag
     4261 cgtcgccagg aaaagatcca ggctgtcctg gaggaccaca gtcagatgag gcagacgcgt
     4321 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     4381 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     4441 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     4501 gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac
     4561 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     4621 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     4681 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     4741 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     4801 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     4861 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     4921 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
     4981 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     5041 gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     5101 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     5161 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5221 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5281 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     5341 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     5401 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     5461 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     5521 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     5581 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     5641 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     5701 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     5761 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5821 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5881 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     5941 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     6001 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     6061 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     6121 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     6181 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     6241 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     6301 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     6361 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     6421 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     6481 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     6541 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     6601 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     6661 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     6721 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     6781 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     6841 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6901 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6961 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     7021 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     7081 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     7141 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     7201 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     7261 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     7321 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     7381 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     7441 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     7501 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     7561 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7621 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7681 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7741 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7801 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     7861 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7921 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7981 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     8041 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     8101 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     8161 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     8221 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     8281 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     8341 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
