LOCUS       sequence-111921-        7788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of PABPC1-MS2 fusion, double mutant..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7788)
  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 7788)
  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 MLLEmut M161A/D165K
FEATURES             Location/Qualifiers
     source          1..7788
                     /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    3492..3613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3503..3522)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3557..3576
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3799..4254
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3886..3905)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4096..4117
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4371..4728
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4371..4562
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4394..4414)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4579..4714
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4779..5573
                     /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(4833..4852)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5443..5462
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5637..5685
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5726..5745)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5845..5867
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5905..5923)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5991..6095
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6096..6164,6165..6956)
                     /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(6314..6333)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7127..7715
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7616..7635
                     /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 tgatggaagg
     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 ggcggcgccg cccgcattga
     3301 taattcagaa cttcttcaca tgctggagtc tccagagtca ctccgttcta aggttgatga
     3361 agctgtagct gtactacaag cccaccaagc taaagaggct gcccagaaag cagttaacag
     3421 tgccaccggt gttccaactg tttaagcggc cgcttccctt tagtgagggt taatgcttcg
     3481 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3541 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3601 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     3661 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcc gataaggatc
     3721 gatccgggct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     3781 agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3841 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3901 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3961 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4021 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4081 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4141 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4201 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat
     4261 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgcg gatctgcgca
     4321 gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
     4381 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4441 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4501 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4561 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4621 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4681 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttgattct
     4741 tctgacacaa cagtctcgaa cttaaggcta gagccaccat gattgaacaa gatggattgc
     4801 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4861 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4921 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4981 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5041 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5101 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5161 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5221 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5281 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5341 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5401 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5461 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5521 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5581 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgat ggccgcaata
     5641 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagcgataa
     5701 ggatccgcgt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     5761 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     5821 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     5881 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     5941 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6001 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     6061 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     6121 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6181 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     6241 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     6301 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     6361 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     6421 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     6481 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     6541 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6601 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     6661 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     6721 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     6781 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     6841 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     6901 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     6961 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     7021 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7081 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     7141 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     7201 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     7261 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     7321 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     7381 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     7441 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     7501 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     7561 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     7621 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     7681 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     7741 ggttcctggc cttttgctgg ccttttgctc acatggctcg acagatct
//
