LOCUS       sequence-289009-        8213 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg6-del577-816.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8213)
  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 8213)
  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-del577-816_I
FEATURES             Location/Qualifiers
     source          1..8213
                     /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      4153..4162
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4159..4161,4162..4164,4165..4878)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4204..4225)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4465..4484)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4812..4833
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5001..5122
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5038..5057)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5092..5111
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5129..5584)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5266..5287)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5478..5497
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5611..5715
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5717..6074
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5717..5908
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5740..5760)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5925..6060
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6109..6903
                     /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             6109..6903
                     /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(6163..6182)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6773..6792
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7091..7110)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7135..7182
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7511..8099
                     /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 ggaacctgac gcgtgcagaa
     2341 cagatggaaa agaagcaaca tgaggaattt gacctgagcc ctgaccagtg gcggaaagga
     2401 aagaagtcta ctttccggca tgttggagat gacaccactc gcctggagat ctggattcat
     2461 ccatcccatc cacggtcttc ccagggcact gagtctggga aggattctga gcaagagaat
     2521 gggctgggca gcctgagtcc cagtgatctg aacaaaaggt tcatcctcag ttttctccat
     2581 gcccatggga agctgtttac ccggattggg atggagacat tccctgcagt ggctgagaag
     2641 gtcctcaagg agttccaggt gttactgcag cacagcccct ctcccattgg aagtacccgc
     2701 atgctgcagc ttatgaccat caatatgttt gcagtacaca actcccagct gaaagactgc
     2761 ttctcggagg agtgccgctc tgtgatccag gaacaagccg cagctctggg cttggccatg
     2821 ttttctctac tggtccgccg ctgcacctgc ttacttaagg agtccgccaa agctcagctg
     2881 tcctctcctg aggaccagga tgaccaagac gacatcaagg tgtcttcctt tgtcccggac
     2941 ctgaaggagc tgctccccag tgtcaaagtc tggtcagatt ggatgctcgg ctacccggac
     3001 acctggaatc ctcctcccac atccctggat ctgccctcgc atgttgctgt ggatgtatgg
     3061 tcgacgctgg ctgatttctg taacatactg actgcagtga atcagtctga ggtgccactg
     3121 tacaaggacc cggatgatga cctcaccctt cttatcctgg aagaggatcg gcttctctcg
     3181 ggctttgtcc ccttgctggc tgcccctcag gacccctgct acgtggagaa aacctcggat
     3241 aaggttattg cagctgactg caaaagggtc acagtgctga agtattttct ggaagccctt
     3301 tgtggacaag aagagcctct gctggcattc aagggtggaa agtatgtgtc agtggcaccc
     3361 gtcccagaca ccatgggaaa ggaaatggga agccaagagg gaacacgact ggaagatgag
     3421 gaggaggatg tggtgattga agactttgag gaagattcag aggctgaagg cagcggaggc
     3481 gaggatgaca tcagggagct tcgggccaag aagctggctc tggccaggaa gatagctgag
     3541 cagcagcgtc gccaggaaaa gatccaggct gtcctggagg accacagtca gatgaggcag
     3601 atggagctcg aaatcagacc tttgttcctc gtaccagaca ccaacggctt cattgaccac
     3661 ctggccagtc tggcgcggct gctggagagc aggaagtaca tcctggtggt gcccctcatc
     3721 gtgatcaatg agctggacgg cctggccaag gggcaggaga cagaccaccg ggctgggggc
     3781 tacgcccgtg tggtacaaga gaaggcccgc aagtccatcg agttcctcga gcagcgattc
     3841 gagagtcggg actcttgcct gcgagccctg accagccgtg gcaatgaact cgaatccatc
     3901 gccttccgca gtgaggacat cactggccag ctgggtaaca acgatgatct catcctgtcc
     3961 tgctgcctcc actactgcaa agacaaggct aaggacttca tgcccgccag caaagaggag
     4021 ccaatccggc tactgcggga ggtggtgctg ttgacggatg accggaacct gcgtgtgaag
     4081 gcgctcacaa ggaatgttcc tgtacgggac atcccagcct tcctcacgtg ggcccaggtg
     4141 ggcaaaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     4201 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     4261 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     4321 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc
     4381 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     4441 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     4501 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     4561 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     4621 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     4681 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     4741 ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag
     4801 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     4861 gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg
     4921 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     4981 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     5041 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5101 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     5161 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     5221 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     5281 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     5341 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     5401 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     5461 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     5521 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     5581 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5641 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5701 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     5761 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5821 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5881 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5941 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     6001 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     6061 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     6121 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     6181 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     6241 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     6301 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     6361 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     6421 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     6481 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     6541 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     6601 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     6661 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     6721 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     6781 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     6841 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     6901 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     6961 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     7021 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     7081 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     7141 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     7201 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     7261 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     7321 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     7381 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     7441 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     7501 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     7561 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     7621 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     7681 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7741 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7801 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7861 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7921 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7981 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     8041 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     8101 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     8161 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
