LOCUS       sequence-289911-        8851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsSmg7-T103E-siRNAres-shRNAres.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8851)
  AUTHORS   Jonas S, Weichenrieder O, Izaurralde E
  TITLE     An unusual arrangement of two 14-3-3-like domains in the SMG5-SMG7 
            heterodimer is required for efficient nonsense-mediated mRNA decay.
  JOURNAL   Genes Dev. 2013 Jan 15;27(2):211-25. doi: 10.1101/gad.206672.112.
  PUBMED    23348841
REFERENCE   2  (bases 1 to 8851)
  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"; doi: 
            "10.1101/gad.206672"; journalName: "Genes Dev"; date: "2013-01-15- 
            15"; volume: "27"; issue: "2"; pages: "211-25"
COMMENT     Sequence Label: pCIneo-lambdaN-HA-HsSmg7-T103E-siRNAres_S
FEATURES             Location/Qualifiers
     source          1..8851
                     /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             1104..1169
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     CDS             1194..1220
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4555..4676
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4566..4585)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4620..4639
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4862..5317
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4949..4968)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5159..5180
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5434..5791
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5434..5625
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5457..5477)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5642..5777
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5842..6636
                     /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(5896..5915)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6506..6525
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6700..6748
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6789..6808)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6908..6930
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6968..6986)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7054..7158
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7159..7227,7228..8019)
                     /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(7377..7396)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8190..8778
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8679..8698
                     /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 accatggacg cacaaacacg acgacgtgag cgtcgcgctg
     1141 agaaacaagc tcaatggaaa gctgcaaacg gtggatctcc tccacatatg gcttacccat
     1201 acgatgttcc agattacgct cctccactcg agatgagcct gcagagcgcg cagtacctcc
     1261 ggcaggcaga agtcctgaag gctgacatga cagattctaa gctgggtcca gctgaagtct
     1321 ggacatccag gcaggctctg caggacctgt accagaaaat gctagttacc gatttggaat
     1381 acgctttaga caagaaagta gaacaggatc tctggaatca cgcctttaag aatcagatca
     1441 caacactaca aggccaggca aagaatcgag caaatccgaa tcggagtgaa gttcaggcaa
     1501 acctttctct gttcctagag gcagctagtg gcttctatga acagttatta caagaactgt
     1561 gtacagtatt taatgtagat ttaccatgcc gtgtgaagtc ttcccaattg ggaattatca
     1621 gcaataaaca gacgcatacc agcgccatag tgaagccaca gtctagctcc tgttcctata
     1681 tctgccagca ctgcctcgtc caccttggag acattgctcg atacagaaac cagaccagcc
     1741 aggcagagtc ctactatagg catgcagctc agcttgtccc ctccaatggt cagccttata
     1801 atcagttggc tatcttagct tcttccaaag gagaccatct gaccacaatt ttctactact
     1861 gcagaagcat tgctgtgaag ttccctttcc cagctgcctc cactaatctg caaaaagcac
     1921 tttctaaagc actggaaagc cgagatgagg tgaaaaccaa gtggggtgtt tctgacttca
     1981 tcaaggcctt tattaaattc cacggtcatg tgtacctgag taagagcttg gaaaagttga
     2041 gccctcttcg agagaaattg gaagaacagt ttaagaggct gctattccaa aaagctttca
     2101 actctcagca gttagttcat gtcactgtca ttaacctgtt tcaacttcat caccttcgtg
     2161 actttagcaa tgaaaccgag cagcacactt atagccaaga tgagcagcta tgttggacac
     2221 agttgctggc cctctttatg tcttttcttg gcatcctgtg caagtgtcct ctacagaatg
     2281 agtctcagga ggagtcctac aatgcctatc ctcttccagc agtcaaggtc tccatggact
     2341 ggctaagact cagacccagg gtctttcagg aggcagtggt ggatgaaaga cagtacattt
     2401 ggccctggtt gatttccctt ctgaatagtt tccatcccca tgaagaggac ctctcaagta
     2461 ttagtgcgac accacttcca gaggagtttg aattacaagg atttttggca ttgagacctt
     2521 ctttcaggaa cttggatttt tccaaaggtc accagggtat tacaggggac aaagaaggcc
     2581 agcaacgacg aatacgacag caacgcttga tctctatagg caaatggatt gctgataatc
     2641 agccaaggct gattcagtgt gaaaatgagg tagggaaatt gttgtttatc acagaaatcc
     2701 cagaattaat actggaagac cccagtgaag ccaaagagaa cctcattctg caggaaacat
     2761 ctgtgataga gtcgctggct gcagatggga gcccagggct aaaatcagtg ctatctacaa
     2821 gccgaaattt aagcaacaac tgtgacacag gagagaagcc agtggttacc ttcaaagaaa
     2881 acattaagac acgagaagtg aacagagacc aaggaagaag ttttcctccc aaagaggtaa
     2941 aatcccagac agaactaaga aagactccag tgtctgaagc cagaaaaaca cctgtaactc
     3001 aaaccccaac tcaagcaagt aactcccagt tcatccccat tcatcaccct ggagccttcc
     3061 ctcctcttcc cagcaggcca gggtttccgc ccccaacata tgttatcccc ccgcctgtgg
     3121 cattttctat gggctcaggt tacaccttcc cagctggtgt ttctgtccca ggaacctttc
     3181 ttcggcctac agctcactct ccagcaggaa accaggtgca agctgggaaa cagtcccaca
     3241 ttccttacag ccagcaacgg ccctctggac cagggccaat gaaccaggga cctcaacaat
     3301 cacagccacc ttcccagcaa ccccttacat ctttaccagc tcagcctact gctcaaagta
     3361 ctagtcaact gcaggttcaa gctctaactc agcaacaaca atcccctaca aaagctgtgc
     3421 cggctttggg gaaaagcccg cctcaccact ctggattcca gcagtatcaa caggcagatg
     3481 cctccaaaca gctgtggaat ccccctcagg ttcaaggccc attagggaaa attatgcctg
     3541 tgaaacagcc ctactacctt cagacccaag accccataaa actgtttgag ccgtcattgc
     3601 aacctcctgt aatgcagcag cagcctctag aaaaaaaaat gaagcctttt cccatggagc
     3661 catataacca taatccctca gaagtcaagg tcccagaatt ctactgggat tcttcctaca
     3721 gcatggctga taacagatct gtaatggcac agcaagcaaa catagaccgc aggggcaaac
     3781 ggtcaccagg aatcttccgt ccagagcagg atcctgtacc cagaatgccg tttgagaaat
     3841 ccttattgga gaagccctca gagctcatgt cacattcatc ctctttcctg tccctcaccg
     3901 gattctctct caatcaggaa agatacccaa ataatagtat gttcaatgag gtatatggga
     3961 aaaacctgac atccagctcc aaagcagaac tcagtccctc aatggccccc caggaaacat
     4021 ctctgtattc cctttttgaa gggactccgt ggtctccatc acttcctgcc agttcagatc
     4081 attcaacacc agccagccag tctcctcatt cctctaaccc aagcagccta cccagctctc
     4141 ctccaacaca caaccataat tctgttccat tctccaattt tggacccatt gggactccag
     4201 ataacaggga tagaaggact gcagatcggt ggaaaactga taagccagcc atgggtgggt
     4261 ttggcattga ttatctctca gcaacgtcat cctctgagag cagttggcat caggccagca
     4321 ctccgagtgg cacctggaca ggccatggcc cttccatgga ggattcctct gctgtcctca
     4381 tggaaagcct aaagtctatc tggtccagtt ccatgatgca tcctggacct tctgctctgg
     4441 agcagctgtt aatgcagcag aagcagaaac agcaacgggg acaaggcacc atgaaccctc
     4501 cacactgagc ggccgcttcc ctttagtgag ggttaatgct tcgagcagac atgataagat
     4561 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     4621 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     4681 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
     4741 gcaagtaaaa cctctacaaa tgtggtaaaa tccgataagg atcgatccgg gctggcgtaa
     4801 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     4861 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4921 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4981 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     5041 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     5101 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     5161 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     5221 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5281 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     5341 cgcatctgtg cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat
     5401 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa
     5461 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5521 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     5581 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     5641 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5701 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     5761 aggctttttt ggaggcctag gcttttgcaa aaagcttgat tcttctgaca caacagtctc
     5821 gaacttaagg ctagagccac catgattgaa caagatggat tgcacgcagg ttctccggcc
     5881 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5941 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     6001 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     6061 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     6121 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     6181 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     6241 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     6301 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     6361 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     6421 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     6481 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     6541 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     6601 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     6661 ccgaccaagc gacgcccaac ctgccatcac gatggccgca ataaaatatc tttattttca
     6721 ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga taaggatccg cgtatggtgc
     6781 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     6841 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     6901 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     6961 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     7021 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     7081 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     7141 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     7201 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     7261 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     7321 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     7381 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     7441 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     7501 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     7561 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     7621 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     7681 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     7741 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     7801 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     7861 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     7921 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     7981 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     8041 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     8101 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     8161 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     8221 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     8281 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     8341 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     8401 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     8461 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     8521 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     8581 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     8641 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     8701 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     8761 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     8821 tggccttttg ctcacatggc tcgacagatc t
//
