LOCUS       sequence-356532-        8317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg6-del36-576-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8317)
  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 8317)
  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-del36-576-siRNAres_I
FEATURES             Location/Qualifiers
     source          1..8317
                     /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    4021..4142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4032..4051)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4086..4105
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4328..4783
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4415..4434)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4625..4646
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4900..5257
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4900..5091
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4923..4943)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5108..5243
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5308..6102
                     /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(5362..5381)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5972..5991
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6166..6214
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6255..6274)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6374..6396
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6434..6452)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6520..6624
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6625..6693,6694..7485)
                     /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(6843..6862)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7656..8244
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8145..8164
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
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 gatggcggaa gggctggagc gtgtgcggat ctccgcgtcg gagctgcgcg
     1381 ggatcctggc tactctggcc ccgcaggccg ggagcagaga aaacatgaag gaattaacgc
     1441 gtcagcaaca ggagctgcac aggcttctcc gggtggctga caaccaggaa ctgcagctca
     1501 gcaacctgct ctccagggac cgcatcagtc cggagggcct ggagaagatg gcgcaactca
     1561 gagctgaact gctgcagcta tatgagcgct gtattctatt agatattgag ttctctgata
     1621 atcagaatgt ggatcagatc ctgtggaaga atgctttcta tcaggtgatt gagaagttca
     1681 ggcaacttgt caaggatccg aatgttgaga acccagaaca gattcggaac agacttttgg
     1741 agctcttgga tgagggtagt gacttctttg atagtttgct tcagaagctg caggttactt
     1801 acaagttcaa actggaagac tacatggatg gtcttgccat tcgcagcaag ccattacgca
     1861 agacagtaaa atatgccttg atcagtgccc agcgatgcat gatatgccaa ggagatattg
     1921 ctaggtaccg ggagcaggca agcgacacgg caaactatgg gaaagcacgc agttggtacc
     1981 tgaaggccca gcacattgct cccaagaatg ggcgccccta taaccagttg gctttgctgg
     2041 cagtgtatac gaggaggaag cttgacgctg tctattacta tatgcgcagt ttagctgcca
     2101 gcaaccctat cctgactgcc aaggagagtc tcatgagctt gtttgaagag accaagcgga
     2161 aggcagaaca gatggaaaag aagcaacatg aggaatttga cctgagccct gaccagtggc
     2221 ggaaaggaaa gaagtctact ttccggcatg ttggagatga caccactcgc ctggagatct
     2281 ggattcatcc atcccatcca cggtcttccc agggcactga gtctgggaag gattctgagc
     2341 aagagaatgg gctgggcagc ctgagtccca gtgatctgaa caaaaggttc atcctcagtt
     2401 ttctccatgc ccatgggaag ctgtttaccc ggattgggat ggagacattc cctgcagtgg
     2461 ctgagaaggt cctcaaggag ttccaggtgt tactgcagca cagcccctct cccattggaa
     2521 gtacccgcat gctgcagctt atgaccatca atatgtttgc agtacacaac tcccagctga
     2581 aagactgctt ctcggaggag tgccgctctg tgatccagga acaagccgca gctctgggct
     2641 tggccatgtt ttctctactg gtccgccgct gcacctgctt acttaaggag tccgccaaag
     2701 ctcagctgtc ctctcctgag gaccaggatg accaagacga catcaaggtg tcttcctttg
     2761 tcccggacct gaaggagctg ctccccagtg tcaaagtctg gtcagattgg atgctcggct
     2821 acccggacac ctggaatcct cctcccacat ccctggatct gccctcgcat gttgctgtgg
     2881 atgtatggtc gacgctggct gatttctgta acatactgac tgcagtgaat cagtctgagg
     2941 tgccactgta caaggacccg gatgatgacc tcacccttct tatcctggaa gaggatcggc
     3001 ttctctcggg ctttgtcccc ttgctggctg cccctcagga cccctgctac gtggagaaaa
     3061 cctcggataa ggttattgca gctgactgca aaagggtcac agtgctgaag tattttctgg
     3121 aagccctttg tggacaagaa gagcctctgc tggcattcaa gggtggaaag tatgtgtcag
     3181 tggcacccgt cccagacacc atgggaaagg aaatgggaag ccaagaggga acacgactgg
     3241 aagatgagga ggaggatgtg gtgattgaag actttgagga agattcagag gctgaaggca
     3301 gcggaggcga ggatgacatc agggagcttc gggccaagaa gctggctctg gccaggaaga
     3361 tagctgagca gcagcgtcgc caggaaaaga tccaggctgt cctggaggac cacagtcaga
     3421 tgaggcagat ggagctcgaa atcagacctt tgttcctcgt accagacacc aacggcttca
     3481 ttgaccacct ggccagtctg gcgcggctgc tggagagcag gaagtacatc ctggtggtgc
     3541 ccctcatcgt gatcaatgag ctggacggcc tggccaaggg gcaggagaca gaccaccggg
     3601 ctgggggcta cgcccgtgtg gtacaagaga aggcccgcaa gtccatcgag ttcctcgagc
     3661 agcgattcga gagtcgggac tcttgcctgc gagccctgac cagccgtggc aatgaactcg
     3721 aatccatcgc cttccgcagt gaggacatca ctggccagct gggtaacaac gatgatctca
     3781 tcctgtcctg ctgcctccac tactgcaaag acaaggctaa ggacttcatg cccgccagca
     3841 aagaggagcc aatccggcta ctgcgggagg tggtgctgtt gacggatgac cggaacctgc
     3901 gtgtgaaggc gctcacaagg aatgttcctg tacgggacat cccagccttc ctcacgtggg
     3961 cccaggtggg ctgagcggcc gcttcccttt agtgagggtt aatgcttcga gcagacatga
     4021 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     4081 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     4141 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     4201 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatccg ataaggatcg atccgggctg
     4261 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     4321 cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4381 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4441 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4501 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4561 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4621 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4681 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4741 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc
     4801 tccttacgca tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc
     4861 tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct
     4921 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4981 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     5041 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     5101 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     5161 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     5221 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttgattctt ctgacacaac
     5281 agtctcgaac ttaaggctag agccaccatg attgaacaag atggattgca cgcaggttct
     5341 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     5401 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     5461 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     5521 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5581 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5641 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5701 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5761 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5821 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5881 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5941 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     6001 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     6061 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     6121 aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa aatatcttta
     6181 ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag gatccgcgta
     6241 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     6301 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     6361 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     6421 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     6481 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     6541 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     6601 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     6661 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     6721 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     6781 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     6841 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     6901 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6961 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     7021 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     7081 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     7141 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     7201 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     7261 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     7321 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     7381 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     7441 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     7501 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     7561 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7621 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7681 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7741 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7801 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7861 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7921 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7981 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     8041 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     8101 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     8161 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     8221 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     8281 ttttgctggc cttttgctca catggctcga cagatct
//
