LOCUS       sequence-269948-        6951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EIF3B (1-464) 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 6951)
  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 6951)
  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-1-464
FEATURES             Location/Qualifiers
     source          1..6951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          185..317
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        362..380
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      384..393
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             390..1325
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1837..1885
                     /note="synthetic polyadenylation signal"
     promoter        1894..2645
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2715..2847
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        2892..2910
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2919..2928
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2925..4577
                     /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             3884..3892
                     /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    4602..4723
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4613..4632)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4667..4686
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4856..4960
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4961..5029,5030..5821)
                     /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(5179..5198)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5992..6580
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6481..6500
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(6709..6951,1..115)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6709..6900
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6732..6752)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6917..6951,1..101)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
       61 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
      121 tgattcttct gacacaacag tctcgaactt aagctgcaga agttggtcgt gaggcactgg
      181 gcaggtaagt atcaaggtta caagacaggt ttaaggagac caatagaaac tgggcttgtc
      241 gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt
      301 tgcctttctc tccacaggtg tccactccca gttcaattac agctcttaag gctagagtac
      361 ttaatacgac tcactatagg ctagccacca tggcttccaa ggtgtacgac cccgagcaac
      421 gcaaacgcat gatcactggg cctcagtggt gggctcgctg caagcaaatg aacgtgctgg
      481 actccttcat caactactat gattccgaga agcacgccga gaacgccgtg atttttctgc
      541 atggtaacgc tgcctccagc tacctgtgga ggcacgtcgt gcctcacatc gagcccgtgg
      601 ctagatgcat catccctgat ctgatcggaa tgggtaagtc cggcaagagc gggaatggct
      661 catatcgcct cctggatcac tacaagtacc tcaccgcttg gttcgagctg ctgaaccttc
      721 caaagaaaat catctttgtg ggccacgact ggggggcttg tctggccttt cactactcct
      781 acgagcacca agacaagatc aaggccatcg tccatgctga gagtgtcgtg gacgtgatcg
      841 agtcctggga cgagtggcct gacatcgagg aggatatcgc cctgatcaag agcgaagagg
      901 gcgagaaaat ggtgcttgag aataacttct tcgtcgagac catgctccca agcaagatca
      961 tgcggaaact ggagcctgag gagttcgctg cctacctgga gccattcaag gagaagggcg
     1021 aggttagacg gcctaccctc tcctggcctc gcgagatccc tctcgttaag ggaggcaagc
     1081 ccgacgtcgt ccagattgtc cgcaactaca acgcctacct tcgggccagc gacgatctgc
     1141 ctaagatgtt catcgagtcc gaccctgggt tcttttccaa cgctattgtc gagggagcta
     1201 agaagttccc taacaccgag ttcgtgaagg tgaagggcct ccacttcagc caggaggacg
     1261 ctccagatga aatgggtaag tacatcaaga gcttcgtgga gcgcgtgctg aagaacgagc
     1321 agtaattcta ggcgatcgct cgagcctgga gcactgtggg gacggactcc gcctgctgtt
     1381 cccgcgctga gctacaggac tcccgagtgt gagccgcggt tcctctgttg cagcgcagcc
     1441 gtgtgtgctg tggagccgag gccgtcctgc aggaagccgc gtgactcccg actcctccct
     1501 gtgctctctg gctctggact gtgactgcgc ctggattctg ccattgcgac acatttttgt
     1561 gcctttcagc ccctggtgtc tgcagtgggg gatttaaggc acccgcttcc acttctttct
     1621 tgtttggagt tttctgttgg aaccgccggc gttggctccg aagacttagc gacgccactg
     1681 gcggcacctt ctcctgcgcc cagtgatgtt tccacggtgc ctgtacacag ccgagcagca
     1741 tttccgttga aggacttgca tccccattgc gggcagtgct ggacgtgtcc cggagaccca
     1801 ccgggagggt ttaaacctag agcggccgct ggccgcaata aaatatcttt attttcatta
     1861 catctgtgtg ttggtttttt gtgtgaggat ctaaatgagt cttcggacct cgcgggggcc
     1921 gcttaagcgg tggttagggt ttgtctgacg cggggggagg gggaaggaac gaaacactct
     1981 cattcggagg cggctcgggg tttggtcttg gtggccacgg gcacgcagaa gagcgccgcg
     2041 atcctcttaa gcaccccccc gccctccgtg gaggcggggg tttggtcggc gggtggtaac
     2101 tggcgggccg ctgactcggg cgggtcgcgc gccccagagt gtgacctttt cggtctgctc
     2161 gcagaccccc gggcggcgcc gccgcggcgg cgacgggctc gctgggtcct aggctccatg
     2221 gggaccgtat acgtggacag gctctggagc atccgcacga ctgcggtgat attaccggag
     2281 accttctgcg ggacgagccg ggtcacgcgg ctgacgcgga gcgtccgttg ggcgacaaac
     2341 accaggacgg ggcacaggta cactatcttg tcacccggag gcgcgaggga ctgcaggagc
     2401 ttcagggagt ggcgcagctg cttcatcccc gtggcccgtt gctcgcgttt gctggcggtg
     2461 tccccggaag aaatatattt gcatgtcttt agttctatga tgacacaaac cccgcccagc
     2521 gtcttgtcat tggcgaattc gaacacgcag atgcagtcgg ggcggcgcgg tcccaggtcc
     2581 acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg agcgaccctg cagcgacccg
     2641 cttaaaagct tgattcttct gacacaacag tctcgaactt aagctgcaga agttggtcgt
     2701 gaggcactgg gcaggtaagt atcaaggtta caagacaggt ttaaggagac caatagaaac
     2761 tgggcttgtc gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg
     2821 acatccactt tgcctttctc tccacaggtg tccactccca gttcaattac agctcttaag
     2881 gctagagtac ttaatacgac tcactatagg ctagcaaagc caccatggcc gatgctaaga
     2941 acattaagaa gggccctgct cccttctacc ctctggagga tggcaccgct ggcgagcagc
     3001 tgcacaaggc catgaagagg tatgccctgg tgcctggcac cattgccttc accgatgccc
     3061 acattgaggt ggacatcacc tatgccgagt acttcgagat gtctgtgcgc ctggccgagg
     3121 ccatgaagag gtacggcctg aacaccaacc accgcatcgt ggtgtgctct gagaactctc
     3181 tgcagttctt catgccagtg ctgggcgccc tgttcatcgg agtggccgtg gcccctgcta
     3241 acgacattta caacgagcgc gagctgctga acagcatggg catttctcag cctaccgtgg
     3301 tgttcgtgtc taagaagggc ctgcagaaga tcctgaacgt gcagaagaag ctgcctatca
     3361 tccagaagat catcatcatg gactctaaga ccgactacca gggcttccag agcatgtaca
     3421 cattcgtgac atctcatctg cctcctggct tcaacgagta cgacttcgtg ccagagtctt
     3481 tcgacaggga caaaaccatt gccctgatca tgaacagctc tgggtctacc ggcctgccta
     3541 agggcgtggc cctgcctcat cgcaccgcct gtgtgcgctt ctctcacgcc cgcgacccta
     3601 ttttcggcaa ccagatcatc cccgacaccg ctattctgag cgtggtgcca ttccaccacg
     3661 gcttcggcat gttcaccacc ctgggctacc tgatttgcgg ctttcgggtg gtgctgatgt
     3721 accgcttcga ggaggagctg ttcctgcgca gcctgcaaga ctacaaaatt cagtctgccc
     3781 tgctggtgcc aaccctgttc agcttcttcg ctaagagcac cctgatcgac aagtacgacc
     3841 tgtctaacct gcacgagatt gcctctggcg gcgccccact gtctaaggag gtgggcgaag
     3901 ccgtggccaa gcgctttcat ctgccaggca tccgccaggg ctacggcctg accgagacaa
     3961 ccagcgccat tctgattacc ccagagggcg acgacaagcc tggcgccgtg ggcaaggtgg
     4021 tgccattctt cgaggccaag gtggtggacc tggacaccgg caagaccctg ggagtgaacc
     4081 agcgcggcga gctgtgtgtg cgcggcccta tgattatgtc cggctacgtg aataaccctg
     4141 aggccacaaa cgccctgatc gacaaggacg gctggctgca ctctggcgac attgcctact
     4201 gggacgagga cgagcacttc ttcatcgtgg accgcctgaa gtctctgatc aagtacaagg
     4261 gctaccaggt ggccccagcc gagctggagt ctatcctgct gcagcaccct aacattttcg
     4321 acgccggagt ggccggcctg cccgacgacg atgccggcga gctgcctgcc gccgtcgtcg
     4381 tgctggaaca cggcaagacc atgaccgaga aggagatcgt ggactatgtg gccagccagg
     4441 tgacaaccgc caagaagctg cgcggcggag tggtgttcgt ggacgaggtg cccaagggcc
     4501 tgaccggcaa gctggacgcc cgcaagatcc gcgagatcct gatcaaggct aagaaaggcg
     4561 gcaagatcgc cgtgtaatgc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     4621 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     4681 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     4741 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     4801 ctacaaatgt ggtaaaatcg ataaggatcc aggtggcact tttcggggaa atgtgcgcgg
     4861 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4921 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4981 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5041 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5101 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5161 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5221 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5281 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5341 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5401 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5461 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5521 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5581 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5641 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5701 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5761 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5821 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5881 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5941 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6001 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6061 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6121 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6181 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6241 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6301 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6361 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6421 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6481 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6541 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6601 tttacggttc ctggcctttt gctggccttt tgctcacatg gctcgacaga tctgcgcagc
     6661 accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa
     6721 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     6781 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     6841 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     6901 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt c
//
