LOCUS       sequence-288998-        9388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg6_1-1238-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9388)
  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 9388)
  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: pCIneo-V5-SBP-HsSmg6_1-1238-siRNAres_I
FEATURES             Location/Qualifiers
     source          1..9388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1098..1107
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1107..1148
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1170..1283
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWGGGHVVEGLAGELEQLRARLEHHPQGQREP"
     polyA_signal    5092..5213
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5103..5122)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5157..5176
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      5399..5854
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5486..5505)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5696..5717
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5971..6328
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5971..6162
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5994..6014)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6179..6314
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6379..7173
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6433..6452)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7043..7062
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7237..7285
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(7326..7345)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7445..7467
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7505..7523)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7591..7695
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7696..7764,7765..8556)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7914..7933)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8727..9315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc aattgcggcc accatgggta agcctatccc taaccctctc ctcggtctcg
     1141 attctacgcc cggaagcggt agcggtacca tggacgagaa gaccaccggc tgggggggcg
     1201 gccacgtggt ggagggcctg gccggcgagc tggagcagct gcgggccagg ctggagcacc
     1261 accctcaggg ccagcgggag ccctccggcg gctgcaagcc cctcgagaat tcacgcgtgg
     1321 tacctctaga catggcggaa gggctggagc gtgtgcggat ctccgcgtcg gagctgcgcg
     1381 ggatcctggc tactctggcc ccgcaggccg ggagcagaga aaacatgaag gaattaaagg
     1441 aggccaggcc gcgcaaagat aacaggcgtc cagatctgga aatctataag cctggccttt
     1501 ctcggctaag gaacaagccc aaaatcaagg aaccccctgg gagtgaggaa ttcaaagatg
     1561 aaattgttaa tgaccgagat tgctctgctg ttgaaaatgg tacacagccc gttaaagatg
     1621 tctgcaagga actgaacaac caagagcaga atggtcctat agacccagaa aataatcggg
     1681 gacaagaatc ctttcctagg actgctggac aagaggatcg tagtctaaaa attatcaaaa
     1741 gaacaaagaa acccgacctg cagatctatc agcctggacg acgtttgcag actgttagca
     1801 aagaatccgc cagtcgggtg gaggaggaag aagtcctcaa ccaggtagaa caactgagag
     1861 tagaggaaga tgagtgtagg ggaaatgttg cgaaggagga agttgcgaat aaaccagaca
     1921 gggccgagat agaaaagagc ccaggtggtg ggagagtagg ggctgcaaaa ggagaaaaag
     1981 gaaagaggat gggaaaaggg gagggggtga gggaaaccca cgacgacccg gcccgcggga
     2041 ggccgggctc cgcaaagcgc tactcccgct cagacaaacg aaggaatcgc taccgcacgc
     2101 gcagcaccag ctcagctggc agcaacaaca gcgctgaggg agctggcctg acggataatg
     2161 gatgtcgccg ccgccgacag gataggacca aggagaggcc accactgaag aagcaagtgt
     2221 ctgtgtcctc aaccgattcc ttagacgagg acagaattga tgagcctgat ggattaggac
     2281 ccaggagaag ttcagaaagg aagagacatt tagaaagaaa ctggtctggc cgtggggagg
     2341 gtgagcagaa aaccagtgct aaagaatatc gaggcactct tcgtgtcact ttcgatgcag
     2401 aagccatgaa caaagagtct cccatggtga ggtcagccag ggatgatatg gatagaggaa
     2461 agcctgacaa aggcttgagc agtgggggca aaggctctga gaagcaggag tccaaaaacc
     2521 cgaaacaaga acttcggggt cgtggtcgtg gcattctgat tttgcctgcc cataccaccc
     2581 tatctgtcaa ttcagcaggt tctccagagt ccgcgccttt gggacctcgg cttttgtttg
     2641 gatctggtag taagggatct cggagttggg gccgtggagg caccacacgc cgattgtggg
     2701 acccaaacaa tcctgatcag aaacctgctc taaagactca gacgccccag ctacatttct
     2761 tggacactga tgatgaagtc agccctacat cttggggtga ctcacgccag gctcaggcat
     2821 cttactataa gtttcaaaac tctgacaacc cctattatta cccccggaca ccaggccctg
     2881 cctcccagta tccctatacg ggctataacc ctctacagta cccagtgggc cctacgaatg
     2941 gtgtgtaccc agggccttac tacccaggct acccgactcc gtcaggacag tatgtgtgta
     3001 gccctctacc taccagcacc atgagtcccg aggaggtaga gcagcacatg aggaacctgc
     3061 agcaacagga gctgcacagg cttctccggg tggctgacaa ccaggaactg cagctcagca
     3121 acctgctctc cagggaccgc atcagtccgg agggcctgga gaagatggcg caactcagag
     3181 ctgaactgct gcagctatat gagcgctgta ttctattaga tattgagttc tctgataatc
     3241 agaatgtgga tcagatcctg tggaagaatg ctttctatca ggtgattgag aagttcaggc
     3301 aacttgtcaa ggatccgaat gttgagaacc cagaacagat tcggaacaga cttttggagc
     3361 tcttggatga gggtagtgac ttctttgata gtttgcttca gaagctgcag gttacttaca
     3421 agttcaaact ggaagactac atggatggtc ttgccattcg cagcaagcca ttacgcaaga
     3481 cagtaaaata tgccttgatc agtgcccagc gatgcatgat atgccaagga gatattgcta
     3541 ggtaccggga gcaggcaagc gacacggcaa actatgggaa agcacgcagt tggtacctga
     3601 aggcccagca cattgctccc aagaatgggc gcccctataa ccagttggct ttgctggcag
     3661 tgtatacgag gaggaagctt gacgctgtct attactatat gcgcagttta gctgccagca
     3721 accctatcct gactgccaag gagagtctca tgagcttgtt tgaagagacc aagcggaagg
     3781 cagaacagat ggaaaagaag caacatgagg aatttgacct gagccctgac cagtggcgga
     3841 aaggaaagaa gtctactttc cggcatgttg gagatgacac cactcgcctg gagatctgga
     3901 ttcatccatc ccatccacgg tcttcccagg gcactgagtc tgggaaggat tctgagcaag
     3961 agaatgggct gggcagcctg agtcccagtg atctgaacaa aaggttcatc ctcagttttc
     4021 tccatgccca tgggaagctg tttacccgga ttgggatgga gacattccct gcagtggctg
     4081 agaaggtcct caaggagttc caggtgttac tgcagcacag cccctctccc attggaagta
     4141 cccgcatgct gcagcttatg accatcaata tgtttgcagt acacaactcc cagctgaaag
     4201 actgcttctc ggaggagtgc cgctctgtga tccaggaaca agccgcagct ctgggcttgg
     4261 ccatgttttc tctactggtc cgccgctgca cctgcttact taaggagtcc gccaaagctc
     4321 agctgtcctc tcctgaggac caggatgacc aagacgacat caaggtgtct tcctttgtcc
     4381 cggacctgaa ggagctgctc cccagtgtca aagtctggtc agattggatg ctcggctacc
     4441 cggacacctg gaatcctcct cccacatccc tggatctgcc ctcgcatgtt gctgtggatg
     4501 tatggtcgac gctggctgat ttctgtaaca tactgactgc agtgaatcag tctgaggtgc
     4561 cactgtacaa ggacccggat gatgacctca cccttcttat cctggaagag gatcggcttc
     4621 tctcgggctt tgtccccttg ctggctgccc ctcaggaccc ctgctacgtg gagaaaacct
     4681 cggataaggt tattgcagct gactgcaaaa gggtcacagt gctgaagtat tttctggaag
     4741 ccctttgtgg acaagaagag cctctgctgg cattcaaggg tggaaagtat gtgtcagtgg
     4801 cacccgtccc agacaccatg ggaaaggaaa tgggaagcca agagggaaca cgactggaag
     4861 atgaggagga ggatgtggtg attgaagact ttgaggaaga ttcagaggct gaaggcagcg
     4921 gaggcgagga tgacatcagg gagcttcggg ccaagaagct ggctctggcc aggaagatag
     4981 ctgagcagca gcgtcgccag gaaaagatcc aggctgtcct ggaggaccac agtcagatga
     5041 ggcaggcggc cgcttccctt tagtgagggt taatgcttcg agcagacatg ataagataca
     5101 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     5161 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     5221 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca
     5281 agtaaaacct ctacaaatgt ggtaaaatcc gataaggatc gatccgggct ggcgtaatag
     5341 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac
     5401 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5461 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5521 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5581 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5641 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5701 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5761 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5821 gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
     5881 atctgtgcgg tatttcacac cgcatacgcg gatctgcgca gcaccatggc ctgaaataac
     5941 ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt
     6001 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     6061 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     6121 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     6181 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6241 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     6301 cttttttgga ggcctaggct tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa
     6361 cttaaggcta gagccaccat gattgaacaa gatggattgc acgcaggttc tccggccgct
     6421 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     6481 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     6541 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     6601 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     6661 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6721 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6781 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6841 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6901 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6961 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     7021 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     7081 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     7141 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     7201 accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt attttcatta
     7261 catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatccgcgt atggtgcact
     7321 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     7381 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     7441 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     7501 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     7561 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     7621 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     7681 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     7741 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     7801 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     7861 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     7921 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     7981 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     8041 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     8101 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     8161 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     8221 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     8281 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     8341 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     8401 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     8461 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     8521 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     8581 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     8641 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     8701 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     8761 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     8821 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     8881 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     8941 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     9001 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     9061 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     9121 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     9181 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     9241 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     9301 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     9361 ccttttgctc acatggctcg acagatct
//
