LOCUS       sequence-289208-        7112 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsGIDRB86-EBMmut.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7112)
  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 7112)
  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: pT7-EGFP-C1-HsGIDRB86-EBMmut_K
FEATURES             Location/Qualifiers
     source          1..7112
                     /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"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(710..731)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(971..990)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1318..1339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3900..4021
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3937..3956)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3991..4010
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4028..4483)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4165..4186)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4377..4396
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4510..4614
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4616..4973
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4616..4807
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4639..4659)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4824..4959
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5008..5802
                     /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             5008..5802
                     /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(5062..5081)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5672..5691
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5990..6009)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6034..6081
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6410..6998
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6899..6918
                     /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 gcagagctct ctggctaact agagaaccca ctgcttactg
      601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      661 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      721 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      781 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      841 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      901 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      961 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1021 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1081 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1141 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1261 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1321 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1381 gtccggactc agatctcgag ctatgcagca agaatcagag agatgcagag ttagagccag
     1441 agaacctgac atggcacttg aagtacctaa agctcgtagg ggtgcagtac tccttaagac
     1501 aggtgatgaa gaagaaagct gtggttcacc taactctgtg gtgaaagaaa agcaaaaaga
     1561 aagttctctc tcccaaaaag aagtctttaa agacaaaccg gaggctcgaa gactaaatat
     1621 caatcctgat agaaaggagc ataattgtag agaagaaaag aaatcttcaa caaaattaag
     1681 aatggacaca tgccttcaaa aaacaaatag agtttgttct aagagaggaa ccactgaatc
     1741 caaagaagta ttatcccaag gacaacagca gggagcccca aatgctgggg ttataactaa
     1801 tgcacctttg cagagacatt ttaaaccaaa gaaggtggag tgtttggaag ttgaaactac
     1861 ggatgtgaca ggacatgaga ggatacttct ttcacaggcc tgtttagaaa tcagcgaggc
     1921 tcaagttcca agcaaaccat tccaaaatgt ggaattctgt gacttcagta ggcatgaacc
     1981 tgatggggaa gcatttgaag acaaagattt ggaaggcaga attgaaactg ataccaaggt
     2041 tttggagata ctatatgagt ttcctagagt ttttagttct gtcatgaaac ctgagaatat
     2101 gattgtacca ataaaactaa gctctgattc tgaaattgta caacaaagca tgcaaacatc
     2161 agatggaata ttgaatccca gcagcggagg catcaccact acttctgttc ctggaagtcc
     2221 agatggtgtc tttgatcaaa cttgcgtaga ttttgaagtt gagagtgtag gtggtatagc
     2281 caatagtaca ggtttcatct tagatcaaaa agatacagat tccattcctg caactatggg
     2341 tcacatctct ctgtcagaga gcacaaatga cactgttagt ccagtaatga ttagagaatg
     2401 tgagaagaat gacagcactg ctgatgagtt acatgtaaag cacgaacctc ctgatacagc
     2461 tgtccttgct catgaaacac atagagatag tggatttaag aatgtaggtg acattaccaa
     2521 taaagcatgt atgatggaca ctacaggtat gtcctgtagt gatcatgtaa ctgttgatag
     2581 cccttatgta gttgcagtta gaatagctga tgagacctct attaatacac gaagtttctc
     2641 aaagtttgta ggaatgagtg cagatgcaac ccctcttcat gtagctagaa gtgggaatga
     2701 cactgaagat ttcagcaacc cttctgcttg ctcagatatt tatggtgaga gtatttcatc
     2761 tcattttaca gagtcaacag gaaagttgat agagagcttg tcagattgtg cttcctcctt
     2821 acctataaaa aagattgctg gtagtaatta taacactttt ttggactctg aactcagtat
     2881 gttaaatggg acaaaagttc tttcagacag tgccgtgggc attgacctgg gtagtactgg
     2941 tgatacaaca gaagcattgc acgaactaag aactgccgaa gagttcaaaa cagaagagca
     3001 agatgactca gggagtatag aatttggtgt atcttttcct gatagggaat catcatctat
     3061 ggaaacatcc atcgaaccaa aagcaactga aacttctcgc acagagggaa ttactgccat
     3121 tgaggagagc tgggagtcta tgtttaacga tgatggtgac tgcctggatc cacgtcttct
     3181 acaagagtta tcagggaata ccaagagcag agagagcatc caggaaccta gatctgatta
     3241 ctacaatcat gaagttcctg atattgacct cagtgattgt gaattcccac atgtcattga
     3301 aatttatgac tttccccaag aatttcgtac tgaagacctt ctatgggttt tctgcagtta
     3361 tcaaaagaaa ggatttgata ttaaatgggt ggatgataca catgccctag gagtattctc
     3421 cagtccaatt acagctcgtg atgcgttggg tattaaacac accatggtga agattcgtcc
     3481 cttgtcacag gccacaagag cagccaaggc caaagctaga gcttatgctg agttcctcca
     3541 gccagcaaag gagcgtcctg agacttcagc agccctagcc agaaggttag tcatcagtgc
     3601 ccttggggtt cgaagtaagc agagcaaaac cgaacgagaa gcagagctca agaaactgca
     3661 agaagccaga gagagaaagc ggttggaagc caagcaacgg gaagacatct gggaaggcag
     3721 agaccagtct acagtttgac aattctgcag tcgacggtac cgcgggcccg ggatccaccg
     3781 gatctagata actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     3841 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     3901 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3961 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4021 aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     4081 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     4141 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     4201 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     4261 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     4321 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     4381 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     4441 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     4501 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4561 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     4621 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4681 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4741 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4801 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4861 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     4921 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     4981 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     5041 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     5101 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     5161 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     5221 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5281 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5341 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5401 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5461 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5521 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5581 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5641 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5701 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5761 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5821 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5881 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5941 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     6001 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     6061 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     6121 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     6181 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     6241 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6301 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6361 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6421 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6481 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6541 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6601 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6661 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6721 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6781 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6841 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6901 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6961 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7021 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     7081 attctgtgga taaccgtatt accgccatgc at
//
