LOCUS       sequence-269947-        6575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EIF3B (465-552) 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 6575)
  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 6575)
  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-465-552
FEATURES             Location/Qualifiers
     source          1..6575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        65..422
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        65..256
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(88..108)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      273..408
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          492..624
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        669..687
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      691..700
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             697..1632
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1768..1816
                     /note="synthetic polyadenylation signal"
     promoter        1825..2576
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2646..2778
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        2823..2841
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2850..2859
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2856..4508
                     /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             3815..3823
                     /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    4533..4654
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4544..4563)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4598..4617
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4787..4891
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4892..4960,4961..5752)
                     /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(5110..5129)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5923..6511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6412..6431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gacagatctg cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta
       61 ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
      121 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
      181 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
      241 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
      301 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
      361 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
      421 aaaagcttga ttcttctgac acaacagtct cgaacttaag ctgcagaagt tggtcgtgag
      481 gcactgggca ggtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg
      541 gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca
      601 tccactttgc ctttctctcc acaggtgtcc actcccagtt caattacagc tcttaaggct
      661 agagtactta atacgactca ctataggcta gccaccatgg cttccaaggt gtacgacccc
      721 gagcaacgca aacgcatgat cactgggcct cagtggtggg ctcgctgcaa gcaaatgaac
      781 gtgctggact ccttcatcaa ctactatgat tccgagaagc acgccgagaa cgccgtgatt
      841 tttctgcatg gtaacgctgc ctccagctac ctgtggaggc acgtcgtgcc tcacatcgag
      901 cccgtggcta gatgcatcat ccctgatctg atcggaatgg gtaagtccgg caagagcggg
      961 aatggctcat atcgcctcct ggatcactac aagtacctca ccgcttggtt cgagctgctg
     1021 aaccttccaa agaaaatcat ctttgtgggc cacgactggg gggcttgtct ggcctttcac
     1081 tactcctacg agcaccaaga caagatcaag gccatcgtcc atgctgagag tgtcgtggac
     1141 gtgatcgagt cctgggacga gtggcctgac atcgaggagg atatcgccct gatcaagagc
     1201 gaagagggcg agaaaatggt gcttgagaat aacttcttcg tcgagaccat gctcccaagc
     1261 aagatcatgc ggaaactgga gcctgaggag ttcgctgcct acctggagcc attcaaggag
     1321 aagggcgagg ttagacggcc taccctctcc tggcctcgcg agatccctct cgttaaggga
     1381 ggcaagcccg acgtcgtcca gattgtccgc aactacaacg cctaccttcg ggccagcgac
     1441 gatctgccta agatgttcat cgagtccgac cctgggttct tttccaacgc tattgtcgag
     1501 ggagctaaga agttccctaa caccgagttc gtgaaggtga agggcctcca cttcagccag
     1561 gaggacgctc cagatgaaat gggtaagtac atcaagagct tcgtggagcg cgtgctgaag
     1621 aacgagcagt aattctaggc gatcgctcga ggcaccgcca tgccttgtac ccccaccgtg
     1681 caggttgtgg ccggttttct ccgcaggttg aacatggaaa taaaagcaaa cttgtatgag
     1741 tttaaaccta gagcggccgc tggccgcaat aaaatatctt tattttcatt acatctgtgt
     1801 gttggttttt tgtgtgagga tctaaatgag tcttcggacc tcgcgggggc cgcttaagcg
     1861 gtggttaggg tttgtctgac gcggggggag ggggaaggaa cgaaacactc tcattcggag
     1921 gcggctcggg gtttggtctt ggtggccacg ggcacgcaga agagcgccgc gatcctctta
     1981 agcacccccc cgccctccgt ggaggcgggg gtttggtcgg cgggtggtaa ctggcgggcc
     2041 gctgactcgg gcgggtcgcg cgccccagag tgtgaccttt tcggtctgct cgcagacccc
     2101 cgggcggcgc cgccgcggcg gcgacgggct cgctgggtcc taggctccat ggggaccgta
     2161 tacgtggaca ggctctggag catccgcacg actgcggtga tattaccgga gaccttctgc
     2221 gggacgagcc gggtcacgcg gctgacgcgg agcgtccgtt gggcgacaaa caccaggacg
     2281 gggcacaggt acactatctt gtcacccgga ggcgcgaggg actgcaggag cttcagggag
     2341 tggcgcagct gcttcatccc cgtggcccgt tgctcgcgtt tgctggcggt gtccccggaa
     2401 gaaatatatt tgcatgtctt tagttctatg atgacacaaa ccccgcccag cgtcttgtca
     2461 ttggcgaatt cgaacacgca gatgcagtcg gggcggcgcg gtcccaggtc cacttcgcat
     2521 attaaggtga cgcgtgtggc ctcgaacacc gagcgaccct gcagcgaccc gcttaaaagc
     2581 ttgattcttc tgacacaaca gtctcgaact taagctgcag aagttggtcg tgaggcactg
     2641 ggcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt
     2701 cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact
     2761 ttgcctttct ctccacaggt gtccactccc agttcaatta cagctcttaa ggctagagta
     2821 cttaatacga ctcactatag gctagcaaag ccaccatggc cgatgctaag aacattaaga
     2881 agggccctgc tcccttctac cctctggagg atggcaccgc tggcgagcag ctgcacaagg
     2941 ccatgaagag gtatgccctg gtgcctggca ccattgcctt caccgatgcc cacattgagg
     3001 tggacatcac ctatgccgag tacttcgaga tgtctgtgcg cctggccgag gccatgaaga
     3061 ggtacggcct gaacaccaac caccgcatcg tggtgtgctc tgagaactct ctgcagttct
     3121 tcatgccagt gctgggcgcc ctgttcatcg gagtggccgt ggcccctgct aacgacattt
     3181 acaacgagcg cgagctgctg aacagcatgg gcatttctca gcctaccgtg gtgttcgtgt
     3241 ctaagaaggg cctgcagaag atcctgaacg tgcagaagaa gctgcctatc atccagaaga
     3301 tcatcatcat ggactctaag accgactacc agggcttcca gagcatgtac acattcgtga
     3361 catctcatct gcctcctggc ttcaacgagt acgacttcgt gccagagtct ttcgacaggg
     3421 acaaaaccat tgccctgatc atgaacagct ctgggtctac cggcctgcct aagggcgtgg
     3481 ccctgcctca tcgcaccgcc tgtgtgcgct tctctcacgc ccgcgaccct attttcggca
     3541 accagatcat ccccgacacc gctattctga gcgtggtgcc attccaccac ggcttcggca
     3601 tgttcaccac cctgggctac ctgatttgcg gctttcgggt ggtgctgatg taccgcttcg
     3661 aggaggagct gttcctgcgc agcctgcaag actacaaaat tcagtctgcc ctgctggtgc
     3721 caaccctgtt cagcttcttc gctaagagca ccctgatcga caagtacgac ctgtctaacc
     3781 tgcacgagat tgcctctggc ggcgccccac tgtctaagga ggtgggcgaa gccgtggcca
     3841 agcgctttca tctgccaggc atccgccagg gctacggcct gaccgagaca accagcgcca
     3901 ttctgattac cccagagggc gacgacaagc ctggcgccgt gggcaaggtg gtgccattct
     3961 tcgaggccaa ggtggtggac ctggacaccg gcaagaccct gggagtgaac cagcgcggcg
     4021 agctgtgtgt gcgcggccct atgattatgt ccggctacgt gaataaccct gaggccacaa
     4081 acgccctgat cgacaaggac ggctggctgc actctggcga cattgcctac tgggacgagg
     4141 acgagcactt cttcatcgtg gaccgcctga agtctctgat caagtacaag ggctaccagg
     4201 tggccccagc cgagctggag tctatcctgc tgcagcaccc taacattttc gacgccggag
     4261 tggccggcct gcccgacgac gatgccggcg agctgcctgc cgccgtcgtc gtgctggaac
     4321 acggcaagac catgaccgag aaggagatcg tggactatgt ggccagccag gtgacaaccg
     4381 ccaagaagct gcgcggcgga gtggtgttcg tggacgaggt gcccaagggc ctgaccggca
     4441 agctggacgc ccgcaagatc cgcgagatcc tgatcaaggc taagaaaggc ggcaagatcg
     4501 ccgtgtaatg cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     4561 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     4621 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     4681 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     4741 tggtaaaatc gataaggatc caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4801 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4861 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4921 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4981 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5041 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5101 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5161 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5221 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5281 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5341 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5401 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5461 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5521 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5581 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5641 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5701 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5761 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5821 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5881 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5941 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6001 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6061 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6121 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6181 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6241 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6301 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6361 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6421 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6481 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6541 cctggccttt tgctggcctt ttgctcacat ggctc
//
