LOCUS       sequence-111913-        8170 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NMD reporter mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8170)
  AUTHORS   Clerici M, Deniaud A, Boehm V, Gehring NH, Schaffitzel C, Cusack S
  TITLE     Structural and functional analysis of the three MIF4G domains of 
            nonsense-mediated decay factor UPF2.
  JOURNAL   Nucleic Acids Res. 2014 Feb;42(4):2673-86. doi: 10.1093/nar/gkt1197.
            Epub 2013 Nov 23.
  PUBMED    24271394
REFERENCE   2  (bases 1 to 8170)
  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: 
            "10.1093/nar/gkt1197"; journalName: "Nucleic Acids Res"; date: 
            "2014-02"; volume: "42"; issue: "4"; pages: "2673-86"
COMMENT     Sequence Label: NG0888 pCI-TPI-PTC48-HBB
FEATURES             Location/Qualifiers
     source          1..8170
                     /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"
     polyA_signal    3874..3995
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3885..3904)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3939..3958
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4181..4636
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4268..4287)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4478..4499
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4753..5110
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4753..4944
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4776..4796)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4961..5096
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5161..5955
                     /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(5215..5234)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5825..5844
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6019..6067
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6108..6127)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6227..6249
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6287..6305)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6373..6477
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6478..6546,6547..7338)
                     /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(6696..6715)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7509..8097
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7998..8017
                     /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 ctcgagaatt cacgcgtggt acctctagac cttcagcgcc tcggctccag
     1141 caccatggcg ccctccagga agttcttcgt tgggggaaac tggaagatga acgggcggaa
     1201 gcagagtctg ggggagctca tcggcactct gaacgcggcc aaggtgccgg ccgacaccgg
     1261 taagccctcg ccgaggaggg gtctggccgg gccggggccg gggccggggc aggagtggca
     1321 gcgccctctc ccgaggcccc gaggccccga ggccggtatc cgcgcggacc tgatgcaggg
     1381 ctgtgggacg agggccgctg gggtccgggc aggggcctcg cagccgcagc cccgtcggtg
     1441 cgtcgagggg gcagggcgga gcacatgatg ccccttggac tatggggcag gtaaggacgt
     1501 tttcttccct tcggcaacct gaacgactcc cgccttccac ggaagggacc gagcccgtgc
     1561 caaacaggct gagcgatttg ggagtgagga gccatcctac cgctttcccc aacctggaaa
     1621 cagcaaagcg caaggcctct gagtcagtta ggtctctgcc acccacgggc aaaggatgct
     1681 ctcctccatc ctccttcctc cctccaccga aatcggagag ccgcgggcct gatccaaaga
     1741 ggcatcccct tctcgttcat tccccagagg cctcaataca aaccccagga gttggcccct
     1801 ctccttttgc tacaaatcct tgccttgcaa aggggaggtg aggatgggct attttagaag
     1861 ggaagcaggg ttgctccctg gagaatgctg agtctgtgag gtgcctatgc cgagaatagc
     1921 tcgaggaaat tggagcccca gctgttaaaa gagcagaggg cagggtgagg gccgtggcct
     1981 ctcaggggta tctggaaggc tcttcgagtt gagtgcagac ccagcctggg ctggaaaatg
     2041 gacaaaggtc atcttgctgg ggtgaaaagg gggagagcag aaccaagaag aagagggtga
     2101 gggctggggg gctccagggc actggttagg aattgtgggg aatgaaggct ttctttagtc
     2161 tcatccccct gtggtaccat cttgtcctca gaggtggttt gtgctccccc tactgcctaa
     2221 atcgacttcg cccggcagaa gctagatccc aagattgctg tggctgcgca gaactgctac
     2281 aaagtgacta atggggcttt tactggggag atcaggtgag atcgaggtgg agaggggtgt
     2341 gtgggaccct tccctcactt tcctcgttga ggggaaagcc acagggtggg ctccctgctg
     2401 aaccttggct tcatctcttc ctttagccct ggcatgatca aagactgcgg agccacgtgg
     2461 gtggtcctgg ggcactcaga gagaaggcat gtctttgggg agtcagatga ggttagtagc
     2521 caagagagaa gataagggat gtctttttcc aagaaggatg tctcaccaag tctgtttctc
     2581 aacagctgat tgggcagaaa gtggcccatg ctctggcaga gggactcgga gtaatcgcct
     2641 gcattgggga gaagctagat gaaagggaag ctggcatcac tgagaaggtt gttttcgagc
     2701 agacaaaggt catcgcaggt atctctggag aaagggacct ttgagcctat ccagggccac
     2761 agagactcag agggtagggt caggccctgg agcctgtctt ggtccccatg ctgatccaga
     2821 aaaggaaaaa ggggaggggg agtgacaatc tttgcttggg gcctatgact tctccagccc
     2881 caaggtagat gccacctgga aatcccccaa tgtccactag ggggcagtag gccaccgttc
     2941 ttcgtactcc ggagaacctg gctggagagc tctttcttgt tcacccttcc ctccatctgt
     3001 atctctgccc tgcagataac gtgaaggact ggagcaaggt cgtcctggcc tatgagcctg
     3061 tgtgggccat tggtactggc aagactgcaa caccccaaca ggtaaccggg cccaggagcc
     3121 ctgccctcat cccagcctgc ctcaataggt ttggacagac acagcccaca tggggcaacc
     3181 ccttatttca aagacacaga gaccttgaac ccagagacag tgacttgtcc aagggcatcc
     3241 agtccagggc ctggcttgga tcagagccct ggtactctga ctcagtcaga aaccacacta
     3301 agtgtccact ggtgccagtg atttttcctc ttagagaggc agaaaaggtc ttacttaggc
     3361 cagcttcttg ttctaggccc aggaagtaca cgagaagctc cgaggatggc tgaagtccaa
     3421 cgtctctgat gcggtggctc agagcacccg tatcatttat ggaggtgagt ggctttggtt
     3481 cccggctgag gtggagtggg ctgaggacta gactgagccc tcggacatgg aggtggggat
     3541 ggggcagact catcccattc ttgaccaagc ccttgttctg ctcccttccc aggctctgtg
     3601 actggggcaa cctgcaagga gctggccagc cagcctgatg tggatggctt ccttgtgggt
     3661 ggtgcttccc tcaagcccat cactttggca aagaattcac cccaccagtg caggctgcct
     3721 atcagaaagt ggtggctggt gtggctaatg ccctggccca caagtatcac taagctcgct
     3781 ttcttgctgt ccaatttcta ttaaaggttc ctttgttccc taagtccgcg gccgcttccc
     3841 tttagtgagg gttaatgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     3901 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     3961 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     4021 tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat
     4081 gtggtaaaat ccgataagga tcgatccggg ctggcgtaat agcgaagagg cccgcaccga
     4141 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca
     4201 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4261 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4321 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4381 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4441 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4501 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4561 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4621 ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4681 accgcatacg cggatctgcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg
     4741 gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     4801 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4861 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4921 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4981 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5041 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5101 cttttgcaaa aagcttgatt cttctgacac aacagtctcg aacttaaggc tagagccacc
     5161 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5221 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5281 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5341 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5401 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5461 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5521 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5581 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5641 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5701 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5761 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5821 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5881 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5941 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     6001 tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt
     6061 ttgtgtgaat cgatagcgat aaggatccgc gtatggtgca ctctcagtac aatctgctct
     6121 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     6181 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     6241 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     6301 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     6361 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6421 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6481 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6541 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6601 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6661 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6721 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6781 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6841 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6901 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6961 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     7021 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     7081 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     7141 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     7201 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     7261 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7321 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7381 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7441 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7501 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7561 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7621 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7681 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7741 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7801 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7861 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7921 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7981 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     8041 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     8101 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatggct
     8161 cgacagatct
//
