LOCUS       sequence-289010-        7667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg6-del817-1238-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7667)
  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 7667)
  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-del817-1238-siRNAres_I
FEATURES             Location/Qualifiers
     source          1..7667
                     /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      3607..3616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3613..3615,3616..3618,3619..4332)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3658..3679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3919..3938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4266..4287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4455..4576
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4492..4511)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4546..4565
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4583..5038)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4720..4741)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4932..4951
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5065..5169
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5171..5528
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5171..5362
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5194..5214)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5379..5514
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5563..6357
                     /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             5563..6357
                     /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(5617..5636)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6227..6246
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6545..6564)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6589..6636
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6965..7553
                     /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 agcggaagac gcgtatggag
     3061 ctcgaaatca gacctttgtt cctcgtacca gacaccaacg gcttcattga ccacctggcc
     3121 agtctggcgc ggctgctgga gagcaggaag tacatcctgg tggtgcccct catcgtgatc
     3181 aatgagctgg acggcctggc caaggggcag gagacagacc accgggctgg gggctacgcc
     3241 cgtgtggtac aagagaaggc ccgcaagtcc atcgagttcc tcgagcagcg attcgagagt
     3301 cgggactctt gcctgcgagc cctgaccagc cgtggcaatg aactcgaatc catcgccttc
     3361 cgcagtgagg acatcactgg ccagctgggt aacaacgatg atctcatcct gtcctgctgc
     3421 ctccactact gcaaagacaa ggctaaggac ttcatgcccg ccagcaaaga ggagccaatc
     3481 cggctactgc gggaggtggt gctgttgacg gatgaccgga acctgcgtgt gaaggcgctc
     3541 acaaggaatg ttcctgtacg ggacatccca gccttcctca cgtgggccca ggtgggcaaa
     3601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     3661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     3721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     3781 ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc
     3841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     4021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     4081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     4141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     4201 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     4261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     4321 ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     4381 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     4441 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4501 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4561 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     4621 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     4681 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     4741 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     4801 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     4861 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     4921 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     4981 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     5041 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5101 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5161 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     5221 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5281 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5341 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5401 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5461 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5521 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     5581 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5641 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5701 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     5761 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5821 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5881 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5941 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     6001 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     6061 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     6121 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     6181 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     6241 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     6301 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     6361 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     6421 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     6481 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     6541 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     6601 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     6661 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     6721 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     6781 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     6841 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6901 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6961 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     7021 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     7081 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     7141 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     7201 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     7261 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     7321 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     7381 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     7441 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     7501 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     7561 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     7621 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
