LOCUS       sequence-111922-        7791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of eRF3a-MS2 fusion. F76A mutation abolishes binding to 
            PABPC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7791)
  AUTHORS   Fatscher T, Boehm V, Weiche B, Gehring NH
  TITLE     The interaction of cytoplasmic poly(A)-binding protein with 
            eukaryotic initiation factor 4G suppresses nonsense-mediated mRNA 
            decay.
  JOURNAL   RNA. 2014 Oct;20(10):1579-92. doi: 10.1261/rna.044933.114. Epub 2014
            Aug 21.
  PUBMED    25147240
REFERENCE   2  (bases 1 to 7791)
  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: 
            "10.1261/rna.044933.114"; journalName: "RNA"; date: "2014-10"; 
            volume: "20"; issue: "10"; pages: "1579-92"
COMMENT     Sequence Label: pCI-MS2V5-eRF3a F76a
FEATURES             Location/Qualifiers
     source          1..7791
                     /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"
     CDS             1097..1483
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2
                     /note="binds to a specific stem-loop structure in the viral
                     RNA (Peabody, 1993)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQNRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             1490..1531
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    3495..3616
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3506..3525)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3560..3579
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3802..4257
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3889..3908)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4099..4120
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4374..4731
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4374..4565
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4397..4417)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4582..4717
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4782..5576
                     /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(4836..4855)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5446..5465
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5640..5688
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5729..5748)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5848..5870
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5908..5926)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5994..6098
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6099..6167,6168..6959)
                     /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(6317..6336)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7130..7718
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7619..7638
                     /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 accatggctt ctaactttac tcagttcgtt ctcgtggaca atggcggaac
     1141 tggcgacgtg actgtcgccc caagcaactt cgctaacggg gtcgctgaat ggatcagctc
     1201 taactcgcgt tcacaggctt acaaagtaac ctgtagcgtt cgtcagagct ctgcgcagaa
     1261 tcgcaaatac accatcaaag tcgaggtgcc taaagtggca acccagactg ttggtggtgt
     1321 agagcttcct gtagccgcat ggcgttcgta cttaaatatg gaactaacca ttccaatttt
     1381 cgccacgaac tccgactgcg agcttattgt taaggcaatg caaggtctcc taaaagatgg
     1441 aaacccgatt ccctcagcaa tcgcagcaaa ctccggcatc tacggcggtg gcaagcccat
     1501 ccctaaccca ctgctgggcc tggacagcac cctcgaggat ccgggcagtg gcggcggcgg
     1561 cggcggcggc ggcggcggcg ggagcagcag cggcagcagc agcagcgact cggcgcctga
     1621 ctgctgggac caggcggaca tggaagcccc cgggccgggc ccttgcggcg gcggcggctc
     1681 cctggcggcg gcggccgagg cccagcggga gaacctcagc gcggccttca gccggcaact
     1741 caacgtcaac gccaagcccg ccgtgcccaa cgtccacgcc gccgagttcg tgccgtcctt
     1801 cctgcggggc ccggcagcgc cgccaccccc agctggcggc gccgccaata accacggagc
     1861 cggcagcggc gcgggaggcc gtgcggcacc tgtggaatcc tctcaagagg aacagtcatt
     1921 gtgtgaaggt tcaaattcag ctgttagcat ggaactttca gaacctattg tagaaaatgg
     1981 agagacagaa atgtctccgg aagaatcatg ggagcacaaa gaagaaataa gtgaagcaga
     2041 gccagggggt ggttccttgg gagatggaag gccgccagag gaaagtgccc atgaaatgat
     2101 ggaggaggaa gaggaaatcc caaaacctaa gtctgtggtt gcaccgccag gtgctcctaa
     2161 gaaagagcat gtaaatgtag tattcattgg gcacgtagat gctggcaagt caaccattgg
     2221 aggacaaata atgtatttga ctggaatggt tgacaaaagg acgcttgaaa agtatgaaag
     2281 agaagctaaa gagaaaaaca gagaaacttg gtacttgtct tgggccttag acacaaatca
     2341 ggaagaacga gacaagggta aaacagtaga agtgggtcgt gcctattttg aaaccgaaaa
     2401 gaagcatttc acaattctag atgcccctgg ccacaagagt tttgtcccaa atatgattgg
     2461 tggtgcctct caagctgatt tggctgtgct ggtaatctca gccaggaaag gagagtttga
     2521 aactggattt gaaaaaggag gacagacaag agaacatgca atgttggcaa agacagcagg
     2581 tgtaaaacac ctaattgtgc taattaataa gatggatgat ccaacagtaa attggagcaa
     2641 tgagagatat gaagaatgta aggagaaact agtgccattt ttgaaaaaag ttggcttcaa
     2701 tcccaaaaag gacattcact ttatgccctg ctcaggactt actggagcaa atctcaaaga
     2761 gcagtcggat ttctgtcctt ggtacattgg attaccgttt attccatatc tggataattt
     2821 gccgaacttc aatagatcag ttgatggacc aatcaggctg ccaattgtgg ataagtacaa
     2881 ggatatgggc actgtggtcc tgggaaagct ggaatcagga tctatttgta aaggccagca
     2941 gcttgtgatg atgccaaaca agcacaacgt ggaagttctt ggaatacttt ccgatgatgt
     3001 agagactgat accgtagccc caggtgaaaa cctcaaaatc agactgaaag gaattgaaga
     3061 agaggagatt cttccagggt ttatactttg tgatcctaat aatctttgtc attctggacg
     3121 cacatttgat gcccagatag tgattataga gcacaaatcc atcatctgcc caggctataa
     3181 tgcggtgctg catattcata cctgtattga ggaggtggaa ataacagcct taatctgctt
     3241 ggtagacaaa aaatcaggag aaaaaagtaa gacccgaccc cgttttgtca aacaagatca
     3301 agtatgcatt gctcgcttaa ggacagcagg aaccatctgc cttgagacct ttaaagactt
     3361 ccctcagatg ggtcgtttca ccttaagaga tgagggtaag accattgcaa ttggaaaagt
     3421 tctgaaactg gttccagaga aagactaagc ggccgcttcc ctttagtgag ggttaatgct
     3481 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3541 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3601 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3661 gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tccgataagg
     3721 atcgatccgg gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     3781 cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3841 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3901 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3961 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4021 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4081 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4141 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4201 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct
     4261 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gcggatctgc
     4321 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc
     4381 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4441 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4501 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4561 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4621 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4681 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttgat
     4741 tcttctgaca caacagtctc gaacttaagg ctagagccac catgattgaa caagatggat
     4801 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4861 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4921 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4981 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5041 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5101 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5161 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5221 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5281 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     5341 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5401 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5461 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5521 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5581 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gatggccgca
     5641 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga
     5701 taaggatccg cgtatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5761 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5821 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5881 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5941 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     6001 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     6061 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     6121 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     6181 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     6241 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     6301 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6361 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6421 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6481 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     6541 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6601 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     6661 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     6721 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6781 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6841 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6901 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6961 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     7021 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     7081 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     7141 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7201 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     7261 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     7321 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7381 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7441 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7501 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7561 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7621 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     7681 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7741 tacggttcct ggccttttgc tggccttttg ctcacatggc tcgacagatc t
//
