LOCUS       sequence-123829-        7793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of PABPC1-MS2 fusion. Mutations abolish binding to eIF4G 
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7793)
  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 7793)
  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-PABPC1 M161A/D165K
FEATURES             Location/Qualifiers
     source          1..7793
                     /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    3497..3618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3508..3527)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3562..3581
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3804..4259
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3891..3910)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4101..4122
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4376..4733
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4376..4567
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4399..4419)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4584..4719
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4784..5578
                     /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(4838..4857)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5448..5467
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5642..5690
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5731..5750)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5850..5872
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5910..5928)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5996..6100
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6101..6169,6170..6961)
                     /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(6319..6338)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7132..7720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7621..7640
                     /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 cctcgagaac cccagtgccc ccagctaccc
     1561 gatggcctcg ctctacgtgg gggacctcca ccccgacgtg accgaggcga tgctctacga
     1621 gaagttcagc ccggccgggc ccatcctctc catccgggtc tgcagggaca tgatcacccg
     1681 ccgctccttg ggctacgcgt atgtgaactt ccagcagccg gcggacgcgg agcgtgcttt
     1741 ggacaccatg aattttgatg ttataaaggg caagccagta cgcatcatgt ggtctcagcg
     1801 tgatccatca cttcgcaaaa gtggagtagg caacatattc attaaaaatc tggacaaatc
     1861 cattgataat aaagcactgt atgatacatt ttctgctttt ggtaacatcc tttcatgtaa
     1921 ggtggtttgt gatgaaaatg gttccaaggg ctatggattt gtacactttg agacgcagga
     1981 agcagctgaa agagctattg aaaaaatgaa tggagcactc ctaaataaac gcaaagtatt
     2041 tgttggacga tttaagtctc gtaaagaacg agaagctgaa cttggagcta gggcaaaaga
     2101 attcaccaat gtttacatca agaattttgg agaagacatg gatgatgagc gccttaagga
     2161 tctctttggc aagtttgggc ctgccttaag tgtgaaagta atgactgatg aaagtggaaa
     2221 atccaaagga tttggatttg taagctttga aaggcatgaa gatgcacaga aagctgtgga
     2281 tgagatgaac ggaaaggagc tcaatggaaa acaaatttat gttggtcgag ctcagaaaaa
     2341 ggtggaacgg cagacggaac ttaagcgcaa atttgaacag atgaaacaag ataggatcac
     2401 cagataccag ggtgttaatc tttatgtgaa aaatcttgat gatggtattg atgatgaacg
     2461 tctccggaaa gagttttctc catttggtac aatcactagt gcaaaggtta tgatggaggg
     2521 tggtcgcagc aaagggtttg gttttgtatg tttctcctcc ccagaagaag ccactaaagc
     2581 agttacagaa atgaacggta gaattgtggc cacaaagcca ttgtatgtag ctttagctca
     2641 gcgcaaagaa gagcgccagg ctcacctcac taaccagtat atgcagagaa tggcaagtgt
     2701 acgagctgtt cccaaccctg taatcaaccc ctaccagcca gcacctcctt caggttactt
     2761 catggcagct atcccacaga ctcagaaccg tgctgcatac tatcctccta gccaaattgc
     2821 tcaactaaga ccaagtcctc gctggactgc tcagggtgcc agacctcatc cattccaaaa
     2881 tatgcccggt gctatccgcc cagctgctcc tagaccacca tttagtacta tgagaccagc
     2941 ttcttcacag gttccacgag tcatgtcaac acagcgtgtt gctaacacat caacacagac
     3001 aatgggtcca cgtcctgcag ctgcagccgc tgcagctact cctgctgtcc gcaccgttcc
     3061 acagtataaa tatgctgcag gagttcgcaa tcctcagcaa catcttaatg cacagccaca
     3121 agttacaatg caacagcctg ctgttcatgt acaaggtcag gaacctttga ctgcttccat
     3181 gttggcatct gcccctcctc aagagcaaaa gcaaatgttg ggtgaacggc tgtttcctct
     3241 tattcaagcc atgcacccta ctcttgctgg taaaatcact ggcatgttgt tggagattga
     3301 taattcagaa cttcttcaca tgctggagtc tccagagtca ctccgttcta aggttgatga
     3361 agctgtagct gtactacaag cccaccaagc taaagaggct gcccagaaag cagttaacag
     3421 tgccaccggt gttccaactg tttaacccgg gcggccgctt ccctttagtg agggttaatg
     3481 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     3541 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3601 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3661 gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatccgataa
     3721 ggatcgatcc gggctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     3781 tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3841 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3901 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3961 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4021 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4081 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4141 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4201 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc
     4261 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgcggatct
     4321 gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag
     4381 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4441 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4501 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4561 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4621 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     4681 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg
     4741 attcttctga cacaacagtc tcgaacttaa ggctagagcc accatgattg aacaagatgg
     4801 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4861 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4921 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     4981 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5041 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5101 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5161 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5221 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5281 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     5341 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5401 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5461 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5521 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5581 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgatggccg
     5641 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagc
     5701 gataaggatc cgcgtatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5761 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5821 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5881 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5941 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     6001 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     6061 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     6121 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     6181 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     6241 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     6301 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     6361 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6421 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6481 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6541 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6601 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6661 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6721 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6781 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6841 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6901 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6961 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     7021 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     7081 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     7141 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     7201 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     7261 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     7321 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     7381 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7441 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7501 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7561 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7621 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7681 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7741 tttacggttc ctggcctttt gctggccttt tgctcacatg gctcgacaga tct
//
