LOCUS       sequence-289008-        7310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg6-del36-576-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7310)
  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 7310)
  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-del36-576-siRNAres_I
FEATURES             Location/Qualifiers
     source          1..7310
                     /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      3250..3259
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3256..3258,3259..3261,3262..3975)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3301..3322)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3562..3581)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3909..3930
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4098..4219
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4135..4154)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4189..4208
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4226..4681)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4363..4384)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4575..4594
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4708..4812
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4814..5171
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4814..5005
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4837..4857)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5022..5157
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5206..6000
                     /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             5206..6000
                     /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(5260..5279)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5870..5889
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6188..6207)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6232..6279
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6608..7196
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7097..7116
                     /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 gctagccaac
      601 atggcggaag ggctggagcg tgtgcggatc tccgcgtcgg agctgcgcgg gatcctggct
      661 actctggccc cgcaggccgg gagcagagaa aacatgaagg aattaacgcg tcagcaacag
      721 gagctgcaca ggcttctccg ggtggctgac aaccaggaac tgcagctcag caacctgctc
      781 tccagggacc gcatcagtcc ggagggcctg gagaagatgg cgcaactcag agctgaactg
      841 ctgcagctat atgagcgctg tattctatta gatattgagt tctctgataa tcagaatgtg
      901 gatcagatcc tgtggaagaa tgctttctat caggtgattg agaagttcag gcaacttgtc
      961 aaggatccga atgttgagaa cccagaacag attcggaaca gacttttgga gctcttggat
     1021 gagggtagtg acttctttga tagtttgctt cagaagctgc aggttactta caagttcaaa
     1081 ctggaagact acatggatgg tcttgccatt cgcagcaagc cattacgcaa gacagtaaaa
     1141 tatgccttga tcagtgccca gcgatgcatg atatgccaag gagatattgc taggtaccgg
     1201 gagcaggcaa gcgacacggc aaactatggg aaagcacgca gttggtacct gaaggcccag
     1261 cacattgctc ccaagaatgg gcgcccctat aaccagttgg ctttgctggc agtgtatacg
     1321 aggaggaagc ttgacgctgt ctattactat atgcgcagtt tagctgccag caaccctatc
     1381 ctgactgcca aggagagtct catgagcttg tttgaagaga ccaagcggaa ggcagaacag
     1441 atggaaaaga agcaacatga ggaatttgac ctgagccctg accagtggcg gaaaggaaag
     1501 aagtctactt tccggcatgt tggagatgac accactcgcc tggagatctg gattcatcca
     1561 tcccatccac ggtcttccca gggcactgag tctgggaagg attctgagca agagaatggg
     1621 ctgggcagcc tgagtcccag tgatctgaac aaaaggttca tcctcagttt tctccatgcc
     1681 catgggaagc tgtttacccg gattgggatg gagacattcc ctgcagtggc tgagaaggtc
     1741 ctcaaggagt tccaggtgtt actgcagcac agcccctctc ccattggaag tacccgcatg
     1801 ctgcagctta tgaccatcaa tatgtttgca gtacacaact cccagctgaa agactgcttc
     1861 tcggaggagt gccgctctgt gatccaggaa caagccgcag ctctgggctt ggccatgttt
     1921 tctctactgg tccgccgctg cacctgctta cttaaggagt ccgccaaagc tcagctgtcc
     1981 tctcctgagg accaggatga ccaagacgac atcaaggtgt cttcctttgt cccggacctg
     2041 aaggagctgc tccccagtgt caaagtctgg tcagattgga tgctcggcta cccggacacc
     2101 tggaatcctc ctcccacatc cctggatctg ccctcgcatg ttgctgtgga tgtatggtcg
     2161 acgctggctg atttctgtaa catactgact gcagtgaatc agtctgaggt gccactgtac
     2221 aaggacccgg atgatgacct cacccttctt atcctggaag aggatcggct tctctcgggc
     2281 tttgtcccct tgctggctgc ccctcaggac ccctgctacg tggagaaaac ctcggataag
     2341 gttattgcag ctgactgcaa aagggtcaca gtgctgaagt attttctgga agccctttgt
     2401 ggacaagaag agcctctgct ggcattcaag ggtggaaagt atgtgtcagt ggcacccgtc
     2461 ccagacacca tgggaaagga aatgggaagc caagagggaa cacgactgga agatgaggag
     2521 gaggatgtgg tgattgaaga ctttgaggaa gattcagagg ctgaaggcag cggaggcgag
     2581 gatgacatca gggagcttcg ggccaagaag ctggctctgg ccaggaagat agctgagcag
     2641 cagcgtcgcc aggaaaagat ccaggctgtc ctggaggacc acagtcagat gaggcagatg
     2701 gagctcgaaa tcagaccttt gttcctcgta ccagacacca acggcttcat tgaccacctg
     2761 gccagtctgg cgcggctgct ggagagcagg aagtacatcc tggtggtgcc cctcatcgtg
     2821 atcaatgagc tggacggcct ggccaagggg caggagacag accaccgggc tgggggctac
     2881 gcccgtgtgg tacaagagaa ggcccgcaag tccatcgagt tcctcgagca gcgattcgag
     2941 agtcgggact cttgcctgcg agccctgacc agccgtggca atgaactcga atccatcgcc
     3001 ttccgcagtg aggacatcac tggccagctg ggtaacaacg atgatctcat cctgtcctgc
     3061 tgcctccact actgcaaaga caaggctaag gacttcatgc ccgccagcaa agaggagcca
     3121 atccggctac tgcgggaggt ggtgctgttg acggatgacc ggaacctgcg tgtgaaggcg
     3181 ctcacaagga atgttcctgt acgggacatc ccagccttcc tcacgtgggc ccaggtgggc
     3241 aaaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3301 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     3361 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     3421 gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac
     3481 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     3541 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     3601 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     3661 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     3721 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     3781 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     3841 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
     3901 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     3961 gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     4021 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     4081 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4141 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4201 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     4261 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     4321 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     4381 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4441 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4501 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4561 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     4621 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     4681 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4741 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4801 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     4861 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4921 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4981 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     5041 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5101 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5161 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     5221 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5281 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5341 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     5401 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     5461 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     5521 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5581 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5641 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5701 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     5761 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5821 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5881 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5941 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6001 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     6061 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     6121 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     6181 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     6241 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     6301 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     6361 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     6421 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     6481 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6541 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6601 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6661 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6721 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     6781 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6841 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6901 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6961 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7021 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7081 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7141 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7201 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     7261 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
