LOCUS       sequence-269950-        6892 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EIF3B (148-552) 3'UTR fragment inserted into the 3'UTR of Renilla 
            luciferase in the psiCHECK3 plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6892)
  AUTHORS   Fischer JW, Busa VF, Shao Y, Leung AKL
  TITLE     Structure-Mediated RNA Decay by UPF1 and G3BP1.
  JOURNAL   Mol Cell. 2020 Apr 2;78(1):70-84.e6. doi: 
            10.1016/j.molcel.2020.01.021. Epub 2020 Feb 3.
  PUBMED    32017897
REFERENCE   2  (bases 1 to 6892)
  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.molcel.2020.01.021"; journalName: "Mol Cell"; date: 
            "2020-04-2- 2"; volume: "78"; issue: "1"; pages: "70-84.e6"
COMMENT     Sequence Label: psiCHECK3-EIF3B-148-552
FEATURES             Location/Qualifiers
     source          1..6892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          108..240
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        285..303
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      307..316
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             313..1248
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1701..1749
                     /note="synthetic polyadenylation signal"
     promoter        1758..2509
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2579..2711
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        2756..2774
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2783..2792
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2789..4441
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /db_xref="GI:42718124"
                     /protein_id="AAS38485.1"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     RBS             3748..3756
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4466..4587
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4477..4496)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4531..4550
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4720..4824
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4825..4893,4894..5685)
                     /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(5043..5062)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5856..6444
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6345..6364
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(6573..6892,1..38)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6573..6764
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6596..6616)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6781..6892,1..24)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttgattct tctgacacaa
       61 cagtctcgaa cttaagctgc agaagttggt cgtgaggcac tgggcaggta agtatcaagg
      121 ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct
      181 tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag
      241 gtgtccactc ccagttcaat tacagctctt aaggctagag tacttaatac gactcactat
      301 aggctagcca ccatggcttc caaggtgtac gaccccgagc aacgcaaacg catgatcact
      361 gggcctcagt ggtgggctcg ctgcaagcaa atgaacgtgc tggactcctt catcaactac
      421 tatgattccg agaagcacgc cgagaacgcc gtgatttttc tgcatggtaa cgctgcctcc
      481 agctacctgt ggaggcacgt cgtgcctcac atcgagcccg tggctagatg catcatccct
      541 gatctgatcg gaatgggtaa gtccggcaag agcgggaatg gctcatatcg cctcctggat
      601 cactacaagt acctcaccgc ttggttcgag ctgctgaacc ttccaaagaa aatcatcttt
      661 gtgggccacg actggggggc ttgtctggcc tttcactact cctacgagca ccaagacaag
      721 atcaaggcca tcgtccatgc tgagagtgtc gtggacgtga tcgagtcctg ggacgagtgg
      781 cctgacatcg aggaggatat cgccctgatc aagagcgaag agggcgagaa aatggtgctt
      841 gagaataact tcttcgtcga gaccatgctc ccaagcaaga tcatgcggaa actggagcct
      901 gaggagttcg ctgcctacct ggagccattc aaggagaagg gcgaggttag acggcctacc
      961 ctctcctggc ctcgcgagat ccctctcgtt aagggaggca agcccgacgt cgtccagatt
     1021 gtccgcaact acaacgccta ccttcgggcc agcgacgatc tgcctaagat gttcatcgag
     1081 tccgaccctg ggttcttttc caacgctatt gtcgagggag ctaagaagtt ccctaacacc
     1141 gagttcgtga aggtgaaggg cctccacttc agccaggagg acgctccaga tgaaatgggt
     1201 aagtacatca agagcttcgt ggagcgcgtg ctgaagaacg agcagtaatt ctaggcgatc
     1261 gctcgagctc ctccctgtgc tctctggctc tggactgtga ctgcgcctgg attctgccat
     1321 tgcgacacat ttttgtgcct ttcagcccct ggtgtctgca gtgggggatt taaggcaccc
     1381 gcttccactt ctttcttgtt tggagttttc tgttggaacc gccggcgttg gctccgaaga
     1441 cttagcgacg ccactggcgg caccttctcc tgcgcccagt gatgtttcca cggtgcctgt
     1501 acacagccga gcagcatttc cgttgaagga cttgcatccc cattgcgggc agtgctggac
     1561 gtgtcccgga gacccaccgg gagggcaccg ccatgccttg tacccccacc gtgcaggttg
     1621 tggccggttt tctccgcagg ttgaacatgg aaataaaagc aaacttgtat gagtttaaac
     1681 ctagagcggc cgctggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     1741 ttttgtgtga ggatctaaat gagtcttcgg acctcgcggg ggccgcttaa gcggtggtta
     1801 gggtttgtct gacgcggggg gagggggaag gaacgaaaca ctctcattcg gaggcggctc
     1861 ggggtttggt cttggtggcc acgggcacgc agaagagcgc cgcgatcctc ttaagcaccc
     1921 ccccgccctc cgtggaggcg ggggtttggt cggcgggtgg taactggcgg gccgctgact
     1981 cgggcgggtc gcgcgcccca gagtgtgacc ttttcggtct gctcgcagac ccccgggcgg
     2041 cgccgccgcg gcggcgacgg gctcgctggg tcctaggctc catggggacc gtatacgtgg
     2101 acaggctctg gagcatccgc acgactgcgg tgatattacc ggagaccttc tgcgggacga
     2161 gccgggtcac gcggctgacg cggagcgtcc gttgggcgac aaacaccagg acggggcaca
     2221 ggtacactat cttgtcaccc ggaggcgcga gggactgcag gagcttcagg gagtggcgca
     2281 gctgcttcat ccccgtggcc cgttgctcgc gtttgctggc ggtgtccccg gaagaaatat
     2341 atttgcatgt ctttagttct atgatgacac aaaccccgcc cagcgtcttg tcattggcga
     2401 attcgaacac gcagatgcag tcggggcggc gcggtcccag gtccacttcg catattaagg
     2461 tgacgcgtgt ggcctcgaac accgagcgac cctgcagcga cccgcttaaa agcttgattc
     2521 ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt
     2581 aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca
     2641 gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt
     2701 tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga gtacttaata
     2761 cgactcacta taggctagca aagccaccat ggccgatgct aagaacatta agaagggccc
     2821 tgctcccttc taccctctgg aggatggcac cgctggcgag cagctgcaca aggccatgaa
     2881 gaggtatgcc ctggtgcctg gcaccattgc cttcaccgat gcccacattg aggtggacat
     2941 cacctatgcc gagtacttcg agatgtctgt gcgcctggcc gaggccatga agaggtacgg
     3001 cctgaacacc aaccaccgca tcgtggtgtg ctctgagaac tctctgcagt tcttcatgcc
     3061 agtgctgggc gccctgttca tcggagtggc cgtggcccct gctaacgaca tttacaacga
     3121 gcgcgagctg ctgaacagca tgggcatttc tcagcctacc gtggtgttcg tgtctaagaa
     3181 gggcctgcag aagatcctga acgtgcagaa gaagctgcct atcatccaga agatcatcat
     3241 catggactct aagaccgact accagggctt ccagagcatg tacacattcg tgacatctca
     3301 tctgcctcct ggcttcaacg agtacgactt cgtgccagag tctttcgaca gggacaaaac
     3361 cattgccctg atcatgaaca gctctgggtc taccggcctg cctaagggcg tggccctgcc
     3421 tcatcgcacc gcctgtgtgc gcttctctca cgcccgcgac cctattttcg gcaaccagat
     3481 catccccgac accgctattc tgagcgtggt gccattccac cacggcttcg gcatgttcac
     3541 caccctgggc tacctgattt gcggctttcg ggtggtgctg atgtaccgct tcgaggagga
     3601 gctgttcctg cgcagcctgc aagactacaa aattcagtct gccctgctgg tgccaaccct
     3661 gttcagcttc ttcgctaaga gcaccctgat cgacaagtac gacctgtcta acctgcacga
     3721 gattgcctct ggcggcgccc cactgtctaa ggaggtgggc gaagccgtgg ccaagcgctt
     3781 tcatctgcca ggcatccgcc agggctacgg cctgaccgag acaaccagcg ccattctgat
     3841 taccccagag ggcgacgaca agcctggcgc cgtgggcaag gtggtgccat tcttcgaggc
     3901 caaggtggtg gacctggaca ccggcaagac cctgggagtg aaccagcgcg gcgagctgtg
     3961 tgtgcgcggc cctatgatta tgtccggcta cgtgaataac cctgaggcca caaacgccct
     4021 gatcgacaag gacggctggc tgcactctgg cgacattgcc tactgggacg aggacgagca
     4081 cttcttcatc gtggaccgcc tgaagtctct gatcaagtac aagggctacc aggtggcccc
     4141 agccgagctg gagtctatcc tgctgcagca ccctaacatt ttcgacgccg gagtggccgg
     4201 cctgcccgac gacgatgccg gcgagctgcc tgccgccgtc gtcgtgctgg aacacggcaa
     4261 gaccatgacc gagaaggaga tcgtggacta tgtggccagc caggtgacaa ccgccaagaa
     4321 gctgcgcggc ggagtggtgt tcgtggacga ggtgcccaag ggcctgaccg gcaagctgga
     4381 cgcccgcaag atccgcgaga tcctgatcaa ggctaagaaa ggcggcaaga tcgccgtgta
     4441 atgcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     4501 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     4561 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     4621 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     4681 atcgataagg atccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4741 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4801 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4861 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4921 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     4981 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5041 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5101 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5161 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5221 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5281 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5341 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5401 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5461 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5521 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5581 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5641 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5701 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5761 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5821 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5881 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5941 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6001 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6061 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6121 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6181 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6241 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6301 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6361 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6421 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6481 ttttgctggc cttttgctca catggctcga cagatctgcg cagcaccatg gcctgaaata
     6541 acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat
     6601 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     6661 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     6721 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     6781 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     6841 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gt
//
