LOCUS       sequence-269944-        6634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EIF3B (1-147) 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 6634)
  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 6634)
  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-1-147
FEATURES             Location/Qualifiers
     source          1..6634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          162..294
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        339..357
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      361..370
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             367..1302
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1497..1545
                     /note="synthetic polyadenylation signal"
     promoter        1554..2305
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2375..2507
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        2552..2570
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2579..2588
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2585..4237
                     /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             3544..3552
                     /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    4262..4383
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4273..4292)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4327..4346
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4516..4620
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4621..4689,4690..5481)
                     /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(4839..4858)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5652..6240
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6141..6160
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(6369..6634,1..92)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6369..6560
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6392..6412)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6577..6634,1..78)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
       61 gaggcttttt tggaggccta ggcttttgca aaaagcttga ttcttctgac acaacagtct
      121 cgaacttaag ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa
      181 gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt
      241 tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc
      301 actcccagtt caattacagc tcttaaggct agagtactta atacgactca ctataggcta
      361 gccaccatgg cttccaaggt gtacgacccc gagcaacgca aacgcatgat cactgggcct
      421 cagtggtggg ctcgctgcaa gcaaatgaac gtgctggact ccttcatcaa ctactatgat
      481 tccgagaagc acgccgagaa cgccgtgatt tttctgcatg gtaacgctgc ctccagctac
      541 ctgtggaggc acgtcgtgcc tcacatcgag cccgtggcta gatgcatcat ccctgatctg
      601 atcggaatgg gtaagtccgg caagagcggg aatggctcat atcgcctcct ggatcactac
      661 aagtacctca ccgcttggtt cgagctgctg aaccttccaa agaaaatcat ctttgtgggc
      721 cacgactggg gggcttgtct ggcctttcac tactcctacg agcaccaaga caagatcaag
      781 gccatcgtcc atgctgagag tgtcgtggac gtgatcgagt cctgggacga gtggcctgac
      841 atcgaggagg atatcgccct gatcaagagc gaagagggcg agaaaatggt gcttgagaat
      901 aacttcttcg tcgagaccat gctcccaagc aagatcatgc ggaaactgga gcctgaggag
      961 ttcgctgcct acctggagcc attcaaggag aagggcgagg ttagacggcc taccctctcc
     1021 tggcctcgcg agatccctct cgttaaggga ggcaagcccg acgtcgtcca gattgtccgc
     1081 aactacaacg cctaccttcg ggccagcgac gatctgccta agatgttcat cgagtccgac
     1141 cctgggttct tttccaacgc tattgtcgag ggagctaaga agttccctaa caccgagttc
     1201 gtgaaggtga agggcctcca cttcagccag gaggacgctc cagatgaaat gggtaagtac
     1261 atcaagagct tcgtggagcg cgtgctgaag aacgagcagt aattctaggc gatcgctcga
     1321 gcctggagca ctgtggggac ggactccgcc tgctgttccc gcgctgagct acaggactcc
     1381 cgagtgtgag ccgcggttcc tctgttgcag cgcagccgtg tgtgctgtgg agccgaggcc
     1441 gtcctgcagg aagccgcgtg actcccgcgt ttaaacctag agcggccgct ggccgcaata
     1501 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaggat ctaaatgagt
     1561 cttcggacct cgcgggggcc gcttaagcgg tggttagggt ttgtctgacg cggggggagg
     1621 gggaaggaac gaaacactct cattcggagg cggctcgggg tttggtcttg gtggccacgg
     1681 gcacgcagaa gagcgccgcg atcctcttaa gcaccccccc gccctccgtg gaggcggggg
     1741 tttggtcggc gggtggtaac tggcgggccg ctgactcggg cgggtcgcgc gccccagagt
     1801 gtgacctttt cggtctgctc gcagaccccc gggcggcgcc gccgcggcgg cgacgggctc
     1861 gctgggtcct aggctccatg gggaccgtat acgtggacag gctctggagc atccgcacga
     1921 ctgcggtgat attaccggag accttctgcg ggacgagccg ggtcacgcgg ctgacgcgga
     1981 gcgtccgttg ggcgacaaac accaggacgg ggcacaggta cactatcttg tcacccggag
     2041 gcgcgaggga ctgcaggagc ttcagggagt ggcgcagctg cttcatcccc gtggcccgtt
     2101 gctcgcgttt gctggcggtg tccccggaag aaatatattt gcatgtcttt agttctatga
     2161 tgacacaaac cccgcccagc gtcttgtcat tggcgaattc gaacacgcag atgcagtcgg
     2221 ggcggcgcgg tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg
     2281 agcgaccctg cagcgacccg cttaaaagct tgattcttct gacacaacag tctcgaactt
     2341 aagctgcaga agttggtcgt gaggcactgg gcaggtaagt atcaaggtta caagacaggt
     2401 ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata
     2461 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg tccactccca
     2521 gttcaattac agctcttaag gctagagtac ttaatacgac tcactatagg ctagcaaagc
     2581 caccatggcc gatgctaaga acattaagaa gggccctgct cccttctacc ctctggagga
     2641 tggcaccgct ggcgagcagc tgcacaaggc catgaagagg tatgccctgg tgcctggcac
     2701 cattgccttc accgatgccc acattgaggt ggacatcacc tatgccgagt acttcgagat
     2761 gtctgtgcgc ctggccgagg ccatgaagag gtacggcctg aacaccaacc accgcatcgt
     2821 ggtgtgctct gagaactctc tgcagttctt catgccagtg ctgggcgccc tgttcatcgg
     2881 agtggccgtg gcccctgcta acgacattta caacgagcgc gagctgctga acagcatggg
     2941 catttctcag cctaccgtgg tgttcgtgtc taagaagggc ctgcagaaga tcctgaacgt
     3001 gcagaagaag ctgcctatca tccagaagat catcatcatg gactctaaga ccgactacca
     3061 gggcttccag agcatgtaca cattcgtgac atctcatctg cctcctggct tcaacgagta
     3121 cgacttcgtg ccagagtctt tcgacaggga caaaaccatt gccctgatca tgaacagctc
     3181 tgggtctacc ggcctgccta agggcgtggc cctgcctcat cgcaccgcct gtgtgcgctt
     3241 ctctcacgcc cgcgacccta ttttcggcaa ccagatcatc cccgacaccg ctattctgag
     3301 cgtggtgcca ttccaccacg gcttcggcat gttcaccacc ctgggctacc tgatttgcgg
     3361 ctttcgggtg gtgctgatgt accgcttcga ggaggagctg ttcctgcgca gcctgcaaga
     3421 ctacaaaatt cagtctgccc tgctggtgcc aaccctgttc agcttcttcg ctaagagcac
     3481 cctgatcgac aagtacgacc tgtctaacct gcacgagatt gcctctggcg gcgccccact
     3541 gtctaaggag gtgggcgaag ccgtggccaa gcgctttcat ctgccaggca tccgccaggg
     3601 ctacggcctg accgagacaa ccagcgccat tctgattacc ccagagggcg acgacaagcc
     3661 tggcgccgtg ggcaaggtgg tgccattctt cgaggccaag gtggtggacc tggacaccgg
     3721 caagaccctg ggagtgaacc agcgcggcga gctgtgtgtg cgcggcccta tgattatgtc
     3781 cggctacgtg aataaccctg aggccacaaa cgccctgatc gacaaggacg gctggctgca
     3841 ctctggcgac attgcctact gggacgagga cgagcacttc ttcatcgtgg accgcctgaa
     3901 gtctctgatc aagtacaagg gctaccaggt ggccccagcc gagctggagt ctatcctgct
     3961 gcagcaccct aacattttcg acgccggagt ggccggcctg cccgacgacg atgccggcga
     4021 gctgcctgcc gccgtcgtcg tgctggaaca cggcaagacc atgaccgaga aggagatcgt
     4081 ggactatgtg gccagccagg tgacaaccgc caagaagctg cgcggcggag tggtgttcgt
     4141 ggacgaggtg cccaagggcc tgaccggcaa gctggacgcc cgcaagatcc gcgagatcct
     4201 gatcaaggct aagaaaggcg gcaagatcgc cgtgtaatgc ggccgcttcg agcagacatg
     4261 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     4321 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     4381 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     4441 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc aggtggcact
     4501 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4561 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4621 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4681 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4741 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4801 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4861 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4921 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4981 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5041 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5101 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5161 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5221 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5281 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5341 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5401 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5461 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5521 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5581 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5641 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5701 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5761 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5821 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5881 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5941 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6001 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6061 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6121 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6181 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6241 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6301 gctcgacaga tctgcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta
     6361 ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     6421 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     6481 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     6541 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     6601 tccgcccatt ctccgcccca tggctgacta attt
//
