LOCUS       sequence-289001-        8662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg6-del817-1238-siRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8662)
  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 8662)
  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-del817-1238-siRNAres_I
FEATURES             Location/Qualifiers
     source          1..8662
                     /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    4366..4487
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4377..4396)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4431..4450
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4673..5128
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4760..4779)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4970..4991
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5245..5602
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5245..5436
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5268..5288)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5453..5588
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5653..6447
                     /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(5707..5726)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6317..6336
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6511..6559
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6600..6619)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6719..6741
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6779..6797)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6865..6969
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6970..7038,7039..7830)
                     /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(7188..7207)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8001..8589
                     /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 tcacgcgtga
     1321 tggcggaagg gctggagcgt gtgcggatct ccgcgtcgga gctgcgcggg atcctggcta
     1381 ctctggcccc gcaggccggg agcagagaaa acatgaagga attaaaggag gccaggccgc
     1441 gcaaagataa caggcgtcca gatctggaaa tctataagcc tggcctttct cggctaagga
     1501 acaagcccaa aatcaaggaa ccccctggga gtgaggaatt caaagatgaa attgttaatg
     1561 accgagattg ctctgctgtt gaaaatggta cacagcccgt taaagatgtc tgcaaggaac
     1621 tgaacaacca agagcagaat ggtcctatag acccagaaaa taatcgggga caagaatcct
     1681 ttcctaggac tgctggacaa gaggatcgta gtctaaaaat tatcaaaaga acaaagaaac
     1741 ccgacctgca gatctatcag cctggacgac gtttgcagac tgttagcaaa gaatccgcca
     1801 gtcgggtgga ggaggaagaa gtcctcaacc aggtagaaca actgagagta gaggaagatg
     1861 agtgtagggg aaatgttgcg aaggaggaag ttgcgaataa accagacagg gccgagatag
     1921 aaaagagccc aggtggtggg agagtagggg ctgcaaaagg agaaaaagga aagaggatgg
     1981 gaaaagggga gggggtgagg gaaacccacg acgacccggc ccgcgggagg ccgggctccg
     2041 caaagcgcta ctcccgctca gacaaacgaa ggaatcgcta ccgcacgcgc agcaccagct
     2101 cagctggcag caacaacagc gctgagggag ctggcctgac ggataatgga tgtcgccgcc
     2161 gccgacagga taggaccaag gagaggccac cactgaagaa gcaagtgtct gtgtcctcaa
     2221 ccgattcctt agacgaggac agaattgatg agcctgatgg attaggaccc aggagaagtt
     2281 cagaaaggaa gagacattta gaaagaaact ggtctggccg tggggagggt gagcagaaaa
     2341 ccagtgctaa agaatatcga ggcactcttc gtgtcacttt cgatgcagaa gccatgaaca
     2401 aagagtctcc catggtgagg tcagccaggg atgatatgga tagaggaaag cctgacaaag
     2461 gcttgagcag tgggggcaaa ggctctgaga agcaggagtc caaaaacccg aaacaagaac
     2521 ttcggggtcg tggtcgtggc attctgattt tgcctgccca taccacccta tctgtcaatt
     2581 cagcaggttc tccagagtcc gcgcctttgg gacctcggct tttgtttgga tctggtagta
     2641 agggatctcg gagttggggc cgtggaggca ccacacgccg attgtgggac ccaaacaatc
     2701 ctgatcagaa acctgctcta aagactcaga cgccccagct acatttcttg gacactgatg
     2761 atgaagtcag ccctacatct tggggtgact cacgccaggc tcaggcatct tactataagt
     2821 ttcaaaactc tgacaacccc tattattacc cccggacacc aggccctgcc tcccagtatc
     2881 cctatacggg ctataaccct ctacagtacc cagtgggccc tacgaatggt gtgtacccag
     2941 ggccttacta cccaggctac ccgactccgt caggacagta tgtgtgtagc cctctaccta
     3001 ccagcaccat gagtcccgag gaggtagagc agcacatgag gaacctgcag caacaggagc
     3061 tgcacaggct tctccgggtg gctgacaacc aggaactgca gctcagcaac ctgctctcca
     3121 gggaccgcat cagtccggag ggcctggaga agatggcgca actcagagct gaactgctgc
     3181 agctatatga gcgctgtatt ctattagata ttgagttctc tgataatcag aatgtggatc
     3241 agatcctgtg gaagaatgct ttctatcagg tgattgagaa gttcaggcaa cttgtcaagg
     3301 atccgaatgt tgagaaccca gaacagattc ggaacagact tttggagctc ttggatgagg
     3361 gtagtgactt ctttgatagt ttgcttcaga agctgcaggt tacttacaag ttcaaactgg
     3421 aagactacat ggatggtctt gccattcgca gcaagccatt acgcaagaca gtaaaatatg
     3481 ccttgatcag tgcccagcga tgcatgatat gccaaggaga tattgctagg taccgggagc
     3541 aggcaagcga cacggcaaac tatgggaaag cacgcagttg gtacctgaag gcccagcaca
     3601 ttgctcccaa gaatgggcgc ccctataacc agttggcttt gctggcagtg tatacgagga
     3661 ggaagcttga cgctgtctat tactatatgc gcagtttagc tgccagcaac cctatcctga
     3721 ctgccaagga gagtctcatg agcttgtttg aagagaccaa gcggaagacg cgtatggagc
     3781 tcgaaatcag acctttgttc ctcgtaccag acaccaacgg cttcattgac cacctggcca
     3841 gtctggcgcg gctgctggag agcaggaagt acatcctggt ggtgcccctc atcgtgatca
     3901 atgagctgga cggcctggcc aaggggcagg agacagacca ccgggctggg ggctacgccc
     3961 gtgtggtaca agagaaggcc cgcaagtcca tcgagttcct cgagcagcga ttcgagagtc
     4021 gggactcttg cctgcgagcc ctgaccagcc gtggcaatga actcgaatcc atcgccttcc
     4081 gcagtgagga catcactggc cagctgggta acaacgatga tctcatcctg tcctgctgcc
     4141 tccactactg caaagacaag gctaaggact tcatgcccgc cagcaaagag gagccaatcc
     4201 ggctactgcg ggaggtggtg ctgttgacgg atgaccggaa cctgcgtgtg aaggcgctca
     4261 caaggaatgt tcctgtacgg gacatcccag ccttcctcac gtgggcccag gtgggctgag
     4321 cggccgcttc cctttagtga gggttaatgc ttcgagcaga catgataaga tacattgatg
     4381 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     4441 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     4501 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa
     4561 acctctacaa atgtggtaaa atccgataag gatcgatccg ggctggcgta atagcgaaga
     4621 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc
     4681 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4741 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4801 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4861 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4921 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4981 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     5041 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5101 tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     5161 gcggtatttc acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga
     5221 aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca
     5281 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5341 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     5401 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     5461 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     5521 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     5581 tggaggccta ggcttttgca aaaagcttga ttcttctgac acaacagtct cgaacttaag
     5641 gctagagcca ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5701 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5761 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5821 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5881 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5941 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     6001 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     6061 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     6121 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     6181 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     6241 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     6301 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     6361 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     6421 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     6481 cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg
     6541 tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc gcgtatggtg cactctcagt
     6601 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     6661 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     6721 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     6781 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     6841 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6901 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6961 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     7021 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     7081 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     7141 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     7201 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     7261 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     7321 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     7381 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     7441 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     7501 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     7561 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     7621 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     7681 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     7741 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7801 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     7861 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7921 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7981 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     8041 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     8101 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     8161 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     8221 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     8281 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     8341 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     8401 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     8461 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     8521 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     8581 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     8641 gctcacatgg ctcgacagat ct
//
