LOCUS       sequence-289006-        5948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg6_817-1238-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5948)
  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 5948)
  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_817-1238_I
FEATURES             Location/Qualifiers
     source          1..5948
                     /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"
     regulatory      1888..1897
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1894..1896,1897..1899,1900..2613)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1939..1960)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2200..2219)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2547..2568
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2736..2857
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2773..2792)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2827..2846
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2864..3319)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3001..3022)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3213..3232
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3346..3450
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3452..3809
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3452..3643
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3475..3495)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3660..3795
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3844..4638
                     /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             3844..4638
                     /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(3898..3917)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4508..4527
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4826..4845)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4870..4917
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5246..5834
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5735..5754
                     /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 gctagcaatt
      601 gcggccacca tggcagaaca gatggaaaag aagcaacatg aggaatttga cctgagccct
      661 gaccagtggc ggaaaggaaa gaagtctact ttccggcatg ttggagatga caccactcgc
      721 ctggagatct ggattcatcc atcccatcca cggtcttccc agggcactga gtctgggaag
      781 gattctgagc aagagaatgg gctgggcagc ctgagtccca gtgatctgaa caaaaggttc
      841 atcctcagtt ttctccatgc ccatgggaag ctgtttaccc ggattgggat ggagacattc
      901 cctgcagtgg ctgagaaggt cctcaaggag ttccaggtgt tactgcagca cagcccctct
      961 cccattggaa gtacccgcat gctgcagctt atgaccatca atatgtttgc agtacacaac
     1021 tcccagctga aagactgctt ctcggaggag tgccgctctg tgatccagga acaagccgca
     1081 gctctgggct tggccatgtt ttctctactg gtccgccgct gcacctgctt acttaaggag
     1141 tccgccaaag ctcagctgtc ctctcctgag gaccaggatg accaagacga catcaaggtg
     1201 tcttcctttg tcccggacct gaaggagctg ctccccagtg tcaaagtctg gtcagattgg
     1261 atgctcggct acccggacac ctggaatcct cctcccacat ccctggatct gccctcgcat
     1321 gttgctgtgg atgtatggtc gacgctggct gatttctgta acatactgac tgcagtgaat
     1381 cagtctgagg tgccactgta caaggacccg gatgatgacc tcacccttct tatcctggaa
     1441 gaggatcggc ttctctcggg ctttgtcccc ttgctggctg cccctcagga cccctgctac
     1501 gtggagaaaa cctcggataa ggttattgca gctgactgca aaagggtcac agtgctgaag
     1561 tattttctgg aagccctttg tggacaagaa gagcctctgc tggcattcaa gggtggaaag
     1621 tatgtgtcag tggcacccgt cccagacacc atgggaaagg aaatgggaag ccaagaggga
     1681 acacgactgg aagatgagga ggaggatgtg gtgattgaag actttgagga agattcagag
     1741 gctgaaggca gcggaggcga ggatgacatc agggagcttc gggccaagaa gctggctctg
     1801 gccaggaaga tagctgagca gcagcgtcgc caggaaaaga tccaggctgt cctggaggac
     1861 cacagtcaga tgaggcagcc accggtcgcc accatggtga gcaagggcga ggagctgttc
     1921 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1981 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     2041 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg
     2101 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2161 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2221 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2281 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2341 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     2401 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2461 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
     2521 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2581 atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca
     2641 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2701 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2761 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2821 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     2881 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2941 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     3001 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     3061 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     3121 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     3181 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     3241 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     3301 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3361 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3421 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     3481 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3541 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3601 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3661 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3721 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3781 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     3841 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3901 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3961 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4021 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4081 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4141 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4201 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4261 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4321 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     4381 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4441 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4501 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4561 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4621 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4681 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4741 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4801 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4861 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     4921 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     4981 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     5041 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     5101 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5161 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5221 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5281 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5341 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     5401 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5461 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5521 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5581 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5641 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5701 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5761 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5821 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5881 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5941 ccatgcat
//
