LOCUS       sequence-289003-        6407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg6_1-576.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6407)
  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 6407)
  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_1-576_I
FEATURES             Location/Qualifiers
     source          1..6407
                     /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      2347..2356
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2353..2355,2356..2358,2359..3072)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2398..2419)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2659..2678)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3006..3027
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3195..3316
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3232..3251)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3286..3305
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3323..3778)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3460..3481)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3672..3691
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3805..3909
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3911..4268
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3911..4102
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3934..3954)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4119..4254
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4303..5097
                     /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             4303..5097
                     /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(4357..4376)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4967..4986
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5285..5304)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5329..5376
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5705..6293
                     /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 gctagcaatt
      601 gcggccacca tggcggaagg gctggagcgt gtgcggatct ccgcgtcgga gctgcgcggg
      661 atcctggcta ctctggcccc gcaggccggg agcagagaaa acatgaagga attaaaggag
      721 gccaggccgc gcaaagataa caggcgtcca gatctggaaa tctataagcc tggcctttct
      781 cggctaagga acaagcccaa aatcaaggaa ccccctggga gtgaggaatt caaagatgaa
      841 attgttaatg accgagattg ctctgctgtt gaaaatggta cacagcccgt taaagatgtc
      901 tgcaaggaac tgaacaacca agagcagaat ggtcctatag acccagaaaa taatcgggga
      961 caagaatcct ttcctaggac tgctggacaa gaggatcgta gtctaaaaat tatcaaaaga
     1021 acaaagaaac ccgacctgca gatctatcag cctggacgac gtttgcagac tgttagcaaa
     1081 gaatccgcca gtcgggtgga ggaggaagaa gtcctcaacc aggtagaaca actgagagta
     1141 gaggaagatg agtgtagggg aaatgttgcg aaggaggaag ttgcgaataa accagacagg
     1201 gccgagatag aaaagagccc aggtggtggg agagtagggg ctgcaaaagg agaaaaagga
     1261 aagaggatgg gaaaagggga gggggtgagg gaaacccacg acgacccggc ccgcgggagg
     1321 ccgggctccg caaagcgcta ctcccgctca gacaaacgaa ggaatcgcta ccgcacgcgc
     1381 agcaccagct cagctggcag caacaacagc gctgagggag ctggcctgac ggataatgga
     1441 tgtcgccgcc gccgacagga taggaccaag gagaggccac cactgaagaa gcaagtgtct
     1501 gtgtcctcaa ccgattcctt agacgaggac agaattgatg agcctgatgg attaggaccc
     1561 aggagaagtt cagaaaggaa gagacattta gaaagaaact ggtctggccg tggggagggt
     1621 gagcagaaaa ccagtgctaa agaatatcga ggcactcttc gtgtcacttt cgatgcagaa
     1681 gccatgaaca aagagtctcc catggtgagg tcagccaggg atgatatgga tagaggaaag
     1741 cctgacaaag gcttgagcag tgggggcaaa ggctctgaga agcaggagtc caaaaacccg
     1801 aaacaagaac ttcggggtcg tggtcgtggc attctgattt tgcctgccca taccacccta
     1861 tctgtcaatt cagcaggttc tccagagtcc gcgcctttgg gacctcggct tttgtttgga
     1921 tctggtagta agggatctcg gagttggggc cgtggaggca ccacacgccg attgtgggac
     1981 ccaaacaatc ctgatcagaa acctgctcta aagactcaga cgccccagct acatttcttg
     2041 gacactgatg atgaagtcag ccctacatct tggggtgact cacgccaggc tcaggcatct
     2101 tactataagt ttcaaaactc tgacaacccc tattattacc cccggacacc aggccctgcc
     2161 tcccagtatc cctatacggg ctataaccct ctacagtacc cagtgggccc tacgaatggt
     2221 gtgtacccag ggccttacta cccaggctac ccgactccgt caggacagta tgtgtgtagc
     2281 cctctaccta ccagcaccat gagtcccgag gaggtagagc agcacatgag gaacctgcca
     2341 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2401 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2461 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2521 ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc
     2581 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     2641 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     2701 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     2761 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     2821 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     2881 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2941 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     3001 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3061 ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     3121 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3181 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3241 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3301 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     3361 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3421 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3481 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3541 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3601 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3661 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3721 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     3781 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3841 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3901 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     3961 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4021 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4081 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4141 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4201 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4261 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4321 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4381 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4441 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     4501 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4561 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4621 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4681 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4741 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4801 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     4861 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4921 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4981 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5041 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5101 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5161 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5221 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     5281 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5341 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     5401 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     5461 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     5521 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     5581 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5641 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5701 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5761 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5821 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     5881 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5941 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6001 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6061 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6121 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6181 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6241 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6301 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6361 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
