LOCUS       sequence-288636-        7823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of BGG-PTC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7823)
  AUTHORS   Jonas S, Weichenrieder O, Izaurralde E
  TITLE     An unusual arrangement of two 14-3-3-like domains in the SMG5-SMG7 
            heterodimer is required for efficient nonsense-mediated mRNA decay.
  JOURNAL   Genes Dev. 2013 Jan 15;27(2):211-25. doi: 10.1101/gad.206672.112.
  PUBMED    23348841
REFERENCE   2  (bases 1 to 7823)
  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: 
            "10.1101/gad.206672"; journalName: "Genes Dev"; date: "2013-01-15- 
            15"; volume: "27"; issue: "2"; pages: "211-25"
COMMENT     Sequence Label: pCIneo-RLuc-BGG-PTC_E
FEATURES             Location/Qualifiers
     source          1..7823
                     /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             1095..2027
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    3527..3648
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3538..3557)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3592..3611
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3834..4289
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3921..3940)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4131..4152
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4406..4763
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4406..4597
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4429..4449)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4614..4749
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4814..5608
                     /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(4868..4887)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5478..5497
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5672..5720
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5761..5780)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5880..5902
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5940..5958)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6026..6130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6131..6199,6200..6991)
                     /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(6349..6368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7162..7750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7651..7670
                     /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 caccatgact tcgaaagttt atgatccaga acaaaggaaa cggatgataa
     1141 ctggtccgca gtggtgggcc agatgtaaac aaatgaatgt tcttgattca tttattaatt
     1201 attatgattc agaaaaacat gcagaaaatg ctgttatttt tttacatggt aacgcggcct
     1261 cttcttattt atggcgacat gttgtgccac atattgagcc agtagcgcgg tgtattatac
     1321 cagaccttat tggtatgggc aaatcaggca aatctggtaa tggttcttat aggttacttg
     1381 atcattacaa atatcttact gcatggtttg aacttcttaa tttaccaaag aagatcattt
     1441 ttgtcggcca tgattggggt gcttgtttgg catttcatta tagctatgag catcaagata
     1501 agatcaaagc aatagttcac gctgaaagtg tagtagatgt gattgaatca tgggatgaat
     1561 ggcctgatat tgaagaggat attgcgttga tcaaatctga agaaggagaa aaaatggttt
     1621 tggagaataa cttcttcgtg gaaaccatgt tgccatcaaa aatcatgaga aagttagaac
     1681 cagaagaatt tgcagcatat cttgaaccat tcaaagagaa aggtgaagtt cgtcgtccaa
     1741 cattatcatg gcctcgtgaa atcccgttag taaaaggtgg taaacctgac gttgtacaaa
     1801 ttgttaggaa ttataatgct tatctacgtg caagtgatga tttaccaaaa atgtttattg
     1861 aatcggaccc aggattcttt tccaatgcta ttgttgaagg tgccaagaag tttcctaata
     1921 ctgaatttgt caaagtaaaa ggtcttcatt tttcgcaaga agatgcacct gatgaaatgg
     1981 gaaaatatat caaatcgttc gttgagcgag ttctcaaaaa tgaacaacct cgagacacca
     2041 tggtgcacct gactcctgag gagaagtctg ccgttactgc cctgtggggc aaggtgaacg
     2101 tggatgaagt tggtggtgag gccctgggca ggttggtatc aaggttacaa gacaggttta
     2161 aggagaccaa tagaaactgg gcatgtggag acagagaaga ctcttgggtt tctgataggc
     2221 actgactctc tctgcctatt ggtctatttt cccaccctta ggctgctggt ggtctaccct
     2281 tggacctaga gattctttga gtcctttggg gatctgtcca ctcctgatgc tgttatgggc
     2341 aaccctaagg tgaaggctca tggcaagaaa gtgctcggtg cctttagtga tggcctggct
     2401 cacctggaca acctcaaggg cacctttgcc gcactgagtg agctgcactg tgacaagctg
     2461 cacgtggatc ctgagaactt cagggtgagt ctatgggacc cttgatgttt tctttcccct
     2521 tcttttctat ggttaagttc atgtcatagg aaggggagaa gtaacagggt acagtttaga
     2581 atgggaaaca gacgaatgat tgcatcagtg tggaagtctc aggatcgttt tagtttcttt
     2641 tatttgctgt tcataacaat tgttttcttt tgtttaattc ttgctttctt tttttttctt
     2701 ctccgcaatt tttactatta tacttaatgc cttaacattg tgtataacaa aaaggaaata
     2761 tctctgagat acattaagta acttaaaaaa aaactttaca cagtctgcct agtacattac
     2821 tatttggaat atatgtgtgc ttatttgcat attcataatc tccctacttt attttctttt
     2881 atttttaatt gatacataat cattatacat atttatgggt taaagtgtaa tgttttaata
     2941 tgtgtacaca tattgaccaa atcagggtaa ttttgcattt gtaattttaa aaaatgcttt
     3001 cttcttttaa tatacttttt tgtttatctt atttctaata ctttccctaa tctctttctt
     3061 tcagggcaat aatgatacaa tgtatcatgc ctctttgcac cattctaaag aataacagtg
     3121 ataatttctg ggttaaggca atagcaatat ttctgcatat aaatatttct gcatataaat
     3181 tgtaactgat gtaagaggtt tcatattgct aatagcagct acaatccagc taccattctg
     3241 cttttatttt atggttggga taaggctgga ttattctgag tccaagctag gcccttttgc
     3301 taatcatgtt catacctctt atcttcctcc cacagctcct gggcaacgtg ctggtctgtg
     3361 tgctggccca tcactttggc aaagaattca ccccaccagt gcaggctgcc tatcagaaag
     3421 tggtggctgg tgtggctaat gccctggccc acaagtatca ctaatctaga gtcgacccgg
     3481 gcggccgctt ccctttagtg agggttaatg cttcgagcag acatgataag atacattgat
     3541 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     3601 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     3661 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa
     3721 aacctctaca aatgtggtaa aatccgataa ggatcgatcc gggctggcgt aatagcgaag
     3781 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc
     3841 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3901 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3961 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4021 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4081 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4141 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4201 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4261 ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
     4321 tgcggtattt cacaccgcat acgcggatct gcgcagcacc atggcctgaa ataacctctg
     4381 aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     4441 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4501 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4561 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4621 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4681 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4741 ttggaggcct aggcttttgc aaaaagcttg attcttctga cacaacagtc tcgaacttaa
     4801 ggctagagcc accatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4861 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4921 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4981 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     5041 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     5101 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     5161 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5221 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5281 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5341 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5401 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5461 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5521 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5581 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5641 gcgacgccca acctgccatc acgatggccg caataaaata tctttatttt cattacatct
     5701 gtgtgttggt tttttgtgtg aatcgatagc gataaggatc cgcgtatggt gcactctcag
     5761 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     5821 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5881 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     5941 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     6001 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6061 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     6241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     6361 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     6421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     6481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     6541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     6601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     7021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     7081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     7321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7801 tgctcacatg gctcgacaga tct
//
