LOCUS       sequence-111917-        9713 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 9713)
  AUTHORS   Boehm V, Haberman N, Ottens F, Ule J, Gehring NH
  TITLE     3' UTR length and messenger ribonucleoprotein composition determine 
            endocleavage efficiencies at termination codons.
  JOURNAL   Cell Rep. 2014 Oct 23;9(2):555-68. doi: 
            10.1016/j.celrep.2014.09.012. Epub 2014 Oct 9.
  PUBMED    25310981
REFERENCE   2  (bases 1 to 9713)
  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: 
            "10.1016/j.celrep.2014.09.012"; journalName: "Cell Rep"; date: 
            "2014-10-23- 23"; volume: "9"; issue: "2"; pages: "555-68"
COMMENT     Sequence Label: NG1301 pCI-TPI-4MS2-SMG5(short)-4H
FEATURES             Location/Qualifiers
     source          1..9713
                     /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    5417..5538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5428..5447)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5482..5501
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      5724..6179
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5811..5830)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6021..6042
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6296..6653
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6296..6487
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6319..6339)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6504..6639
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6704..7498
                     /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(6758..6777)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7368..7387
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7562..7610
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(7651..7670)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7770..7792
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7830..7848)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7916..8020
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8021..8089,8090..8881)
                     /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(8239..8258)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9052..9640
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9541..9560
                     /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 gcctcggctc cagcaccatg gcgccctcca ggaagttctt cgttggggga
     1141 aactggaaga tgaacgggcg gaagcagagt ctgggggagc tcatcggcac tctgaacgcg
     1201 gccaaggtgc cggccgacac cggtaagccc tcgccgagga ggggtctggc cgggccgggg
     1261 ccggggccgg ggcaggagtg gcagcgccct ctcccgaggc cccgaggccc cgaggccggt
     1321 atccgcgcgg acctgatgca gggctgtggg acgagggccg ctggggtccg ggcaggggcc
     1381 tcgcagccgc agccccgtcg gtgcgtcgag ggggcagggc ggagcacatg atgccccttg
     1441 gactatgggg caggtaagga cgttttcttc ccttcggcaa cctgaacgac tcccgccttc
     1501 cacggaaggg accgagcccg tgccaaacag gctgagcgat ttgggagtga ggagccatcc
     1561 taccgctttc cccaacctgg aaacagcaaa gcgcaaggcc tctgagtcag ttaggtctct
     1621 gccacccacg ggcaaaggat gctctcctcc atcctccttc ctccctccac cgaaatcgga
     1681 gagccgcggg cctgatccaa agaggcatcc ccttctcgtt cattccccag aggcctcaat
     1741 acaaacccca ggagttggcc cctctccttt tgctacaaat ccttgccttg caaaggggag
     1801 gtgaggatgg gctattttag aagggaagca gggttgctcc ctggagaatg ctgagtctgt
     1861 gaggtgccta tgccgagaat aggtcgagga aattggagcc ccagctgtta aaagagcaga
     1921 gggcagggtg agggccgtgg cctctcaggg gtatctggaa ggctcttcga gttgagtgca
     1981 gacccagcct gggctggaaa atggacaaag gtcatcttgc tggggtgaaa agggggagag
     2041 cagaaccaag aagaagaggg tgagggctgg ggggctccag ggcactggtt aggaattgtg
     2101 gggaatgaag gctttcttta gtctcatccc cctgtggtac catcttgtcc tcagaggtgg
     2161 tttgtgctcc ccctactgcc tatatcgact tcgcccggca gaagctagat cccaagattg
     2221 ctgtggctgc gcagaactgc tacaaagtga ctaatggggc ttttactggg gagatcaggt
     2281 gagatcgagg tggagagggg tgtgtgggac ccttccctca ctttcctcgt tgaggggaaa
     2341 gccacagggt gggctccctg ctgaaccttg gcttcatctc ttcctttagc cctggcatga
     2401 tcaaagactg cggagccacg tgggtggtcc tggggcactc agagagaagg catgtctttg
     2461 gggagtcaga tgaggttagt agccaagaga gaagataagg gatgtctttt tccaagaagg
     2521 atgtctcacc aagtctgttt ctcaacagct gattgggcag aaagtggccc atgctctggc
     2581 agagggactc ggagtaatcg cctgcattgg ggagaagcta gatgaaaggg aagctggcat
     2641 cactgagaag gttgttttcg agcagacaaa ggtcatcgca ggtatctctg gagaaaggga
     2701 cctttgagcc tatccagggc cacagagact cagagggtag ggtcaggccc tggagcctgt
     2761 cttggtcccc atgctgatcc agaaaaggaa aaaggggagg gggagtgaca atctttgctt
     2821 ggggcctatg acttctccag ccccaaggta gatgccacct ggaaatcccc caatgtccac
     2881 tagggggcag taggccaccg ttcttcgtac tccggagaac ctggctggag agctctttct
     2941 tgttcaccct tccctccatc tgtatctctg ccctgcagat aacgtgaagg actggagcaa
     3001 ggtcgtcctg gcctatgagc ctgtgtgggc cattggtact ggcaagactg caacacccca
     3061 acaggtaacc gggcccagga gccctgccct catcccagcc tgcctcaata ggtttggaca
     3121 gacacagccc acatggggca accccttatt tcaaagacac agagaccttg aacccagaga
     3181 cagtgacttg tccaagggca tccagtccag ggcctggctt ggatcagagc cctggtactc
     3241 tgactcagtc agaaaccaca ctaagtgtcc actggtgcca gtgatttttc ctcttagaga
     3301 ggcagaaaag gtcttactta ggccagcttc ttgttctagg cccaggaagt acacgagaag
     3361 ctccgaggat ggctgaagtc caacgtctct gatgcggtgg ctcagagcac ccgtatcatt
     3421 tatggaggtg agtggctttg gttcccggct gaggtggagt gggctgagga ctagactgag
     3481 ccctcggaca tggaggtggg gatggggcag actcatccca ttcttgacca agcccttgtt
     3541 ctgctccctt cccaggctct gtgactgggg caacctgcaa ggagctggcc agccagcctg
     3601 atgtggatgg cttccttgtg ggtggtgctt ccctcaagcc cgaattcgtg gacatcatca
     3661 atgccaaaca atgagcccca tccatcttcc ctacccttcc tgccaagcca gggactcgag
     3721 tattgcctag aaagcacgag catcagccgt gcctccaggt cgaatcttca aacgacgacg
     3781 atcacgcgtc gctgcagtgc aggagtcgag tattgcctag aaagcacgag catcagccgt
     3841 gcctccaggt cgaatcttca aacgacgacg atcacgcgtc gctgcagtgc aggagtcgag
     3901 ttggttgata ctgaccccca ggccctgcag tggggctgac tccagatctc tcctgccctc
     3961 cctggcagcc aggaccagca cctgtagtca ccccaccaca cgcagactca tgcacgcaca
     4021 caggagggag gcctagctgc tcagaggctg cagggagggc ccaggagccg gctgggaggg
     4081 tggggtccct ttgttgccaa gacgttagga aagcgaggaa agtgcttgga ttaggagagt
     4141 cttgtgggcc cctggccagc cttcctgcct cagctcccct gctgtctcca ggggcaggtg
     4201 gtaggcatgg gtacctgcat ttcactggag tgggttcttg gatctctgag gggaaggaac
     4261 agcaaaagag gcccttcttc ctcacccaag atgcagggtg gttggggcca ggagtttgga
     4321 ccctctaggt cttgggggaa gagctgggta atacctggtg tctgagtgat tctctgcaga
     4381 cccttcccct cctcaaggat cacccatcct cctttcagcc ccctttatgg ggaccaggca
     4441 gctctggagc cagccacagg ggctgttaga gaagcaaggc ctggagtggc ctgcaccgag
     4501 tagcagggtc agggttcgtg tgctcctcct cctgctgcag gggctgcaca tcccattgcc
     4561 ccacttctgc tttgtgtctc cctctgtcta gcttccaggg cagggagcag gccccaccta
     4621 gggctgcagg cagtctggcc tgtgccagca cggtctcctg tgcccaccag ccccacaggt
     4681 gctgtgcttt gtgctcttgg ctgctgtgct gggacagaat gggatgccag gaagagaaga
     4741 aagggggtgc agtctgaggc caccaccccc cttcctatct aagggagggc tgaagacaag
     4801 gggccggcat tcagtgggca gcagaaagga gaggcccctt gaagctgctc agtcagaggc
     4861 ccccgtccct ccttttgcct tccgcaggac tgaagacctg aaggggctgg cttttggagt
     4921 gttgaggtga atatctgtcg agcaccccac cagtgcaggc tgcctatcag aaagtggtgg
     4981 ctggtgtggc taatgccctg gcccacaagt atcactaagc tcgctttctt gctgtccaat
     5041 ttgtcgagca ccccaccagt gcaggctgcc tatcagaaag tggtggctgg tgtggctaat
     5101 gccctggccc acaagtatca ctaagctcgc tttcttgctg tccaatttgt cgagcacccc
     5161 accagtgcag gctgcctatc agaaagtggt ggctggtgtg gctaatgccc tggcccacaa
     5221 gtatcactaa gctcgctttc ttgctgtcca atttgtcgag caccccacca gtgcaggctg
     5281 cctatcagaa agtggtggct ggtgtggcta atgccctggc ccacaagtat cactaagctc
     5341 gctttcttgc tgtccaattt gtcgacccgg gcggccgctt ccctttagtg agggttaatg
     5401 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     5461 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     5521 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     5581 gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatccgataa
     5641 ggatcgatcc gggctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     5701 tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5761 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5821 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5881 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     5941 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     6001 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     6061 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     6121 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc
     6181 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgcggatct
     6241 gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag
     6301 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     6361 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     6421 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     6481 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     6541 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     6601 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg
     6661 attcttctga cacaacagtc tcgaacttaa ggctagagcc accatgattg aacaagatgg
     6721 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     6781 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     6841 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     6901 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     6961 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     7021 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     7081 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     7141 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     7201 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     7261 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     7321 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     7381 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     7441 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     7501 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgatggccg
     7561 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagc
     7621 gataaggatc cgcgtatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     7681 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     7741 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     7801 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     7861 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     7921 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     7981 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     8041 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     8101 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     8161 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     8221 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     8281 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     8341 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     8401 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     8461 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     8521 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     8581 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     8641 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     8701 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     8761 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     8821 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     8881 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     8941 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     9001 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     9061 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     9121 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     9181 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     9241 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     9301 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     9361 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     9421 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     9481 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     9541 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     9601 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     9661 tttacggttc ctggcctttt gctggccttt tgctcacatg gctcgacaga tct
//
