LOCUS       sequence-289019-        8594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsUPF2-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8594)
  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 8594)
  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-HsUPF2-siRNAres_I
FEATURES             Location/Qualifiers
     source          1..8594
                     /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"
     RBS             2156..2167
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     RBS             2929..2937
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    5382..5503
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5419..5438)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5473..5492
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5510..5965)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5647..5668)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5859..5878
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5992..6096
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6098..6455
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6098..6289
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6121..6141)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6306..6441
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6490..7284
                     /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             6490..7284
                     /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(6544..6563)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7154..7173
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7472..7491)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7516..7563
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7892..8480
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8381..8400
                     /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 ctatgccagc tgagcgtaaa aagccagcaa gtatggaaga
     1441 aaaagactct ttaccaaaca acaaggaaaa agactgcagt gaaaggcgga cagtgagcag
     1501 caaggagagg ccaaaagacg atatcaagct cactgccaag aaggaggtca gcaaggcccc
     1561 tgaagacaag aagaagagac tggaagatga taagagaaaa aaggaagata aagagcggaa
     1621 gaaaaaagac gaagaaaagg tgaaggcaga ggaagaatca aagaaaaaag aagaggaaga
     1681 aaaaaagaaa catcaagagg aagagagaaa gaagcaagaa gagcaggcca aacgtcagca
     1741 agaagaagaa gcagctgctc agatgaaaga aaaagaagaa tccattcagc ttcatcagga
     1801 agcttgggaa cgacatcatt taagaaagga acttcgtagc aaaaaccaaa atgctccgga
     1861 cagccgacca gaggaaaact tcttcagccg cctcgactca agtttgaaga aaaatactgc
     1921 ttttgtcaag aaactaaaaa ctattacaga acaacagaga gactccttgt cccatgattt
     1981 taatggccta aatttaagca aatacattgc agaagctgta gcttccatcg tggaagcaaa
     2041 actaaaaatc tctgatgtga actgtgctgt gcacctctgc tctctctttc accagcgtta
     2101 tgctgacttt gccccatcac ttcttcaggt ctggaaaaaa cattttgaag caaggaaaga
     2161 ggagaaaaca cctaacatca ccaagttaag aactgatttg cgttttattg cagaattgac
     2221 aatagttggg attttcactg acaaggaagg tctttcctta atctatgaac agctaaaaaa
     2281 tattattaat gctgatcggg agtcccacac tcatgtctct gtagtgatta gtttctgtcg
     2341 acattgtgga gatgatattg ctggacttgt accaaggaaa gtaaagagtg ctgcagagaa
     2401 gtttaatttg agttttcctc ctagtgagat aattagtcca gagaaacaac agcccttcca
     2461 gaatctttta aaagagtact ttacgtcttt gaccaaacac ctgaaaaggg accacaggga
     2521 gctccagaat actgagagac aaaacaggcg cattctacat tctaaagggg agctcagtga
     2581 agatagacat aaacagtatg aggaatttgc tatgtcttac cagaagctgc tggcaaattc
     2641 tcaatcctta gcagaccttt tggatgaaaa tatgccagat cttcctcaag acaaaccaac
     2701 accagaagaa catgggcctg gaattgatat attcacacct ggtaaacctg gagaatatga
     2761 cttggaaggt ggtatatggg aagatgaaga tgctcggaat ttttatgaga acctcattga
     2821 tttgaaggct tttgtcccag ccatcttgtt taaagacaat gaaaaaagtt gtcagaataa
     2881 agagtccaac aaagatgata ccaaagaggc aaaagaatct aaggagaata aggaggtatc
     2941 aagtcccgat gatttggaac ttgagttgga gaatctagaa attaatgatg acaccttaga
     3001 attagagggt ggagatgaag ctgaagatct tacaaagaaa cttcttgatg aacaagaaca
     3061 agaagatgag gaagccagca ctggatctca tctcaagctc atagtagatg ctttcctaca
     3121 gcagttaccc aactgtgtca accgagatct gatagacaag gcagcaatgg atttttgcat
     3181 gaacatgaac acaaaagcaa acaggaagaa gttggtacgg gcactcttca tagttcctag
     3241 acaaaggttg gatttgctac cattttatgc aagattggtt gctacattgc atccctgcat
     3301 gtctgatgta gcagaggatc tttgttccat gctgaggggg gatttcagat ttcatgtacg
     3361 gaaaaaggac cagatcaata ttgaaacaaa gaataaaact gttcgtttta taggagaact
     3421 aactaagttt aagatgttca ccaaaaatga cacactgcat tgtttaaaga tgcttctgtc
     3481 agacttctct catcaccata ttgaaatggc atgcaccctg ctggagacat gtggacggtt
     3541 tcttttcaga tctccagaat ctcacctgag gaccagtgta cttttggagc aaatgatgag
     3601 aaagaagcaa gcaatgcatc ttgatgcgag atacgtcaca atggtagaga atgcatatta
     3661 ctactgcaac ccacctccag ctgaaaaaac cgtgaaaaag aaacgtcctc ctctccagga
     3721 atatgtccgg aaacttttgt acaaggatct ctctaaggtt accaccgaga aggttttgag
     3781 acagatgcga aagctgccct ggcaggacca agaagtgaaa gactatgtta tttgttgtat
     3841 gataaacatc tggaatgtga aatataatag tattcattgt gtagccaacc tcttagcagg
     3901 actagtgctc taccaagagg atgttgggat ccacgttgtg gatggagtgt tagaagatat
     3961 tcgattagga atggaggtta atcaacctaa atttaatcag aggcgcatca gcagtgccaa
     4021 gttcttagga gaactttaca attaccgaat ggtggaatca gctgttattt tcagaactct
     4081 gtattctttt acctcatttg gtgttaatcc tgatggctct ccaagttccc tggacccacc
     4141 tgagcatctt ttcagaatta gactcgtatg cactattctg gacacatgtg gccagtactt
     4201 tgacagaggt tccagtaaac gaaaacttga ttgtttcctt gtatattttc agcgttatgt
     4261 ttggtggaag aaaagtttgg aggtttggac aaaagaccat ccatttccta ttgatataga
     4321 ttacatgatc agtgatacac tagaactgct aagaccaaag atcaaactct gtaattctct
     4381 ggaagaatcc atcaggcagg tacaagactt ggaacgagaa ttcttaataa aactaggcct
     4441 agtaaatgac aaagactcaa aagattctat gacagaagga gaaaatcttg aagaggatga
     4501 agaagaagaa gaaggtgggg ctgaaacaga agaacaatct ggaaatgaaa gtgaagtaaa
     4561 tgagccagaa gaagaggagg gttctgataa tgatgatgat gagggagaag aagaggagga
     4621 agagaataca gattacctta cagattccaa taaggaaaat gaaaccgatg aagagaatac
     4681 tgaggtaatg attaaaggcg gtggacttaa gcatgtacct tgtgtagaag atgaggactt
     4741 cattcaagct ctggataaaa tgatgctaga aaatctacag caacgaagtg gtgagtctgt
     4801 taaagtgcac caactagatg ttgccattcc tttgcatctc aaaagccagc tgaggaaagg
     4861 gcccccactg ggaggtgggg aaggagaggc tgagtctgca gacacaatgc cgtttgtcat
     4921 gttaacaaga aaaggcaata aacagcagtt taagatcctt aatgtaccca tgtcctctca
     4981 acttgctgca aatcactgga accagcaaca ggcagaacaa gaagagagga tgagaatgaa
     5041 aaagctcaca ctagatatca atgaacggca agaacaagaa gattatcaag aaatgttgca
     5101 gtctcttgca cagcgcccag ctccagcaaa caccaatcgt gagaggcggc ctcgctacca
     5161 acatccgaag ggagcaccta atgcagatct aatctttaag actggtggga ggagacgttg
     5221 acaattctgc agtcgacggt accgcgggcc cgggatccac cggatctaga taactgatca
     5281 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     5341 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     5401 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5461 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc
     5521 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     5581 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     5641 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     5701 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     5761 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     5821 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     5881 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     5941 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     6001 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     6061 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     6121 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     6181 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     6241 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     6301 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     6361 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     6421 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     6481 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     6541 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     6601 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     6661 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     6721 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     6781 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     6841 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     6901 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     6961 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     7021 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     7081 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     7141 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     7201 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     7261 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     7321 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     7381 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     7441 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     7501 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     7561 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     7621 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     7681 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     7741 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7801 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7861 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7921 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7981 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     8041 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     8101 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     8161 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     8221 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     8281 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     8341 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     8401 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     8461 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     8521 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     8581 ttaccgccat gcat
//
