LOCUS       sequence-289904-        7225 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian Expression of CeSmg5-G91E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7225)
  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 7225)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-CeSmg5-G91E_S
FEATURES             Location/Qualifiers
     source          1..7225
                     /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    2929..3050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2940..2959)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2994..3013
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3236..3691
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3323..3342)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3533..3554
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3808..4165
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3808..3999
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3831..3851)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4016..4151
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4216..5010
                     /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(4270..4289)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4880..4899
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5074..5122
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5163..5182)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5282..5304
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5342..5360)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5428..5532
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5533..5601,5602..6393)
                     /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(5751..5770)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6564..7152
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7053..7072
                     /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 agatgcaaaa gtcagatgaa gtcactgaaa
     1261 agttcaaacg atattgtaat caactggaaa aatacggcca aactgaaaat gtgcattctc
     1321 cagtgatggc gatgttgcgc agaaaaggcc gaaaacagct cattgagatt atgaaacgcg
     1381 acggggactg tacaagttca atcaataaat tgtggattgt tggatattat caccctttcc
     1441 agttttttat tcgagataag gagaaaaaca tggcaatagc cgtgttgctg acaatgttct
     1501 gtgaagagct tcaagaaatg ctttcgcttc ccgatgacaa atacccagca ttgtggaaca
     1561 tgtacattgg agattttcat cgttatatgc cagacgaaga aattcagaaa tgcttggcag
     1621 ttggatatta ttcaagagca atcgatctag atccgaatca aggaagagct ttccacgttc
     1681 tcgctggtct ccgagctgac ctgaatgttg cacagaagct gagactgatg attttaggac
     1741 agctcgccga tgctccatat aaaaaaggca ctgaactctt ggaatatttg aaatttccac
     1801 aaaaagaatc cactgataaa ttgatggtag attttgtcat ttgggcattg aacgagaaat
     1861 caaaacgaat ggattatcag atgactggaa tcaaaatagt aaacgagttt aaagcagaaa
     1921 tcgaacagaa actcgaattc gattggtctc tgatcatgtc aacgtgtcgt ttagcatcta
     1981 aattggcaat gaagaaattt ggatttcaac aattctacaa ttgcttcgac acaatttcaa
     2041 cgctgtatat tacaatttat tcacgtacga tttcatccaa atgccttcta gctgaagcta
     2101 tctcatggat ttccgattca gccgaaatac tcggacactt ggatgagcag aaaaatgagc
     2161 cacatttcca gaagctatcg gttttcgcca aaacgaaatg gaacgagctc aacgacttgg
     2221 ttatgaatca tatcaattcc gtattcacat ccatgtcact cacaatcaat ccatctattt
     2281 ctatgaccag ctttctgctg aatggtccaa tatccgaacc caatgttgaa tttttgagtc
     2341 aattaatcaa ttatctggtt tccgtcgaat tcccaccaat ggagataatc cacgatagag
     2401 aagaatcggg tccattgcta cgacgaatta atcaatcgga acaaaaacgt ttggatattc
     2461 aaataaagac acaaaatgat gaagttaacc gggaggattg gagacctgtt tatgtattga
     2521 tggattatga cgtcatagtt gacaaaataa gaattgctct aaaaatttgg gatatcgatg
     2581 atttcatctg cattttgcca tcaacagtgc ttgatgaact tgattatcag aaaacgaaga
     2641 atcgagcagt tcgtccagta attcgggcat taatggagct tcaagcagag ggaaaaattg
     2701 tgctaaagaa atgtgataac gagagaagct gcgcggagca attggttcaa agtgcacgaa
     2761 gatctgcaga agatcacaag catattgtcg cttttctgtg caaaaacccg agtgaacaga
     2821 aagaaatgga aggagttact ttttacgata tcgatcagtt ctacatgaag tatttggagt
     2881 gagcggccgc ttccctttag tgagggttaa tgcttcgagc agacatgata agatacattg
     2941 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     3001 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     3061 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt
     3121 aaaacctcta caaatgtggt aaaatccgat aaggatcgat ccgggctggc gtaatagcga
     3181 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
     3241 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3301 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3361 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3421 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3481 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3541 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3601 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3661 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     3721 tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca ccatggcctg aaataacctc
     3781 tgaaagagga acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg
     3841 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3901 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3961 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4021 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4081 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     4141 ttttggaggc ctaggctttt gcaaaaagct tgattcttct gacacaacag tctcgaactt
     4201 aaggctagag ccaccatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4261 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4321 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4381 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4441 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4501 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4561 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4621 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4681 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4741 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4801 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4861 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4921 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4981 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5041 aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt ttcattacat
     5101 ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tccgcgtatg gtgcactctc
     5161 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5221 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5281 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5341 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5401 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5461 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5521 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5581 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5641 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5701 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5761 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5821 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5881 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5941 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6001 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6061 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6121 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6181 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6241 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6301 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6361 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6421 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6481 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6541 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6601 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6661 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6721 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6781 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6841 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6901 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6961 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7021 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7081 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7141 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7201 tttgctcaca tggctcgaca gatct
//
