LOCUS       sequence-269764-        6529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EIF3B (465-552) 3'UTR with only the main hairpin 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 6529)
  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 6529)
  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-88nt-1st-Hairpin
FEATURES             Location/Qualifiers
     source          1..6529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        155..512
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        155..346
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(178..198)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      363..498
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          582..714
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        759..777
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      781..790
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             787..1722
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1812..1860
                     /note="synthetic polyadenylation signal"
     promoter        1869..2620
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2690..2822
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        2867..2885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2894..2903
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2900..4552
                     /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             3859..3867
                     /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    4577..4698
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4588..4607)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4642..4661
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4831..4935
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4936..5004,5005..5796)
                     /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(5154..5173)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5967..6529,1..26)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6456..6475
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
       61 ccttttgctg gccttttgct cacatggctc gacagatctg cgcagcacca tggcctgaaa
      121 taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga
      181 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
      241 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
      301 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
      361 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
      421 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
      481 gaggcttttt tggaggccta ggcttttgca aaaagcttga ttcttctgac acaacagtct
      541 cgaacttaag ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa
      601 gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt
      661 tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc
      721 actcccagtt caattacagc tcttaaggct agagtactta atacgactca ctataggcta
      781 gccaccatgg cttccaaggt gtacgacccc gagcaacgca aacgcatgat cactgggcct
      841 cagtggtggg ctcgctgcaa gcaaatgaac gtgctggact ccttcatcaa ctactatgat
      901 tccgagaagc acgccgagaa cgccgtgatt tttctgcatg gtaacgctgc ctccagctac
      961 ctgtggaggc acgtcgtgcc tcacatcgag cccgtggcta gatgcatcat ccctgatctg
     1021 atcggaatgg gtaagtccgg caagagcggg aatggctcat atcgcctcct ggatcactac
     1081 aagtacctca ccgcttggtt cgagctgctg aaccttccaa agaaaatcat ctttgtgggc
     1141 cacgactggg gggcttgtct ggcctttcac tactcctacg agcaccaaga caagatcaag
     1201 gccatcgtcc atgctgagag tgtcgtggac gtgatcgagt cctgggacga gtggcctgac
     1261 atcgaggagg atatcgccct gatcaagagc gaagagggcg agaaaatggt gcttgagaat
     1321 aacttcttcg tcgagaccat gctcccaagc aagatcatgc ggaaactgga gcctgaggag
     1381 ttcgctgcct acctggagcc attcaaggag aagggcgagg ttagacggcc taccctctcc
     1441 tggcctcgcg agatccctct cgttaaggga ggcaagcccg acgtcgtcca gattgtccgc
     1501 aactacaacg cctaccttcg ggccagcgac gatctgccta agatgttcat cgagtccgac
     1561 cctgggttct tttccaacgc tattgtcgag ggagctaaga agttccctaa caccgagttc
     1621 gtgaaggtga agggcctcca cttcagccag gaggacgctc cagatgaaat gggtaagtac
     1681 atcaagagct tcgtggagcg cgtgctgaag aacgagcagt aattctaggc gatcgctcga
     1741 ggcaccgcca tgccttgtac ccccaccgtg caggttgtgg ccggtttaaa cctagagcgg
     1801 ccgctggccg caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg
     1861 aggatctaaa tgagtcttcg gacctcgcgg gggccgctta agcggtggtt agggtttgtc
     1921 tgacgcgggg ggagggggaa ggaacgaaac actctcattc ggaggcggct cggggtttgg
     1981 tcttggtggc cacgggcacg cagaagagcg ccgcgatcct cttaagcacc cccccgccct
     2041 ccgtggaggc gggggtttgg tcggcgggtg gtaactggcg ggccgctgac tcgggcgggt
     2101 cgcgcgcccc agagtgtgac cttttcggtc tgctcgcaga cccccgggcg gcgccgccgc
     2161 ggcggcgacg ggctcgctgg gtcctaggct ccatggggac cgtatacgtg gacaggctct
     2221 ggagcatccg cacgactgcg gtgatattac cggagacctt ctgcgggacg agccgggtca
     2281 cgcggctgac gcggagcgtc cgttgggcga caaacaccag gacggggcac aggtacacta
     2341 tcttgtcacc cggaggcgcg agggactgca ggagcttcag ggagtggcgc agctgcttca
     2401 tccccgtggc ccgttgctcg cgtttgctgg cggtgtcccc ggaagaaata tatttgcatg
     2461 tctttagttc tatgatgaca caaaccccgc ccagcgtctt gtcattggcg aattcgaaca
     2521 cgcagatgca gtcggggcgg cgcggtccca ggtccacttc gcatattaag gtgacgcgtg
     2581 tggcctcgaa caccgagcga ccctgcagcg acccgcttaa aagcttgatt cttctgacac
     2641 aacagtctcg aacttaagct gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
     2701 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
     2761 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     2821 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     2881 ataggctagc aaagccacca tggccgatgc taagaacatt aagaagggcc ctgctccctt
     2941 ctaccctctg gaggatggca ccgctggcga gcagctgcac aaggccatga agaggtatgc
     3001 cctggtgcct ggcaccattg ccttcaccga tgcccacatt gaggtggaca tcacctatgc
     3061 cgagtacttc gagatgtctg tgcgcctggc cgaggccatg aagaggtacg gcctgaacac
     3121 caaccaccgc atcgtggtgt gctctgagaa ctctctgcag ttcttcatgc cagtgctggg
     3181 cgccctgttc atcggagtgg ccgtggcccc tgctaacgac atttacaacg agcgcgagct
     3241 gctgaacagc atgggcattt ctcagcctac cgtggtgttc gtgtctaaga agggcctgca
     3301 gaagatcctg aacgtgcaga agaagctgcc tatcatccag aagatcatca tcatggactc
     3361 taagaccgac taccagggct tccagagcat gtacacattc gtgacatctc atctgcctcc
     3421 tggcttcaac gagtacgact tcgtgccaga gtctttcgac agggacaaaa ccattgccct
     3481 gatcatgaac agctctgggt ctaccggcct gcctaagggc gtggccctgc ctcatcgcac
     3541 cgcctgtgtg cgcttctctc acgcccgcga ccctattttc ggcaaccaga tcatccccga
     3601 caccgctatt ctgagcgtgg tgccattcca ccacggcttc ggcatgttca ccaccctggg
     3661 ctacctgatt tgcggctttc gggtggtgct gatgtaccgc ttcgaggagg agctgttcct
     3721 gcgcagcctg caagactaca aaattcagtc tgccctgctg gtgccaaccc tgttcagctt
     3781 cttcgctaag agcaccctga tcgacaagta cgacctgtct aacctgcacg agattgcctc
     3841 tggcggcgcc ccactgtcta aggaggtggg cgaagccgtg gccaagcgct ttcatctgcc
     3901 aggcatccgc cagggctacg gcctgaccga gacaaccagc gccattctga ttaccccaga
     3961 gggcgacgac aagcctggcg ccgtgggcaa ggtggtgcca ttcttcgagg ccaaggtggt
     4021 ggacctggac accggcaaga ccctgggagt gaaccagcgc ggcgagctgt gtgtgcgcgg
     4081 ccctatgatt atgtccggct acgtgaataa ccctgaggcc acaaacgccc tgatcgacaa
     4141 ggacggctgg ctgcactctg gcgacattgc ctactgggac gaggacgagc acttcttcat
     4201 cgtggaccgc ctgaagtctc tgatcaagta caagggctac caggtggccc cagccgagct
     4261 ggagtctatc ctgctgcagc accctaacat tttcgacgcc ggagtggccg gcctgcccga
     4321 cgacgatgcc ggcgagctgc ctgccgccgt cgtcgtgctg gaacacggca agaccatgac
     4381 cgagaaggag atcgtggact atgtggccag ccaggtgaca accgccaaga agctgcgcgg
     4441 cggagtggtg ttcgtggacg aggtgcccaa gggcctgacc ggcaagctgg acgcccgcaa
     4501 gatccgcgag atcctgatca aggctaagaa aggcggcaag atcgccgtgt aatgcggccg
     4561 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     4621 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     4681 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     4741 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     4801 gatccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4861 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4921 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4981 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5041 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5101 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5161 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5221 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5281 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5341 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5401 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5461 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5521 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5581 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5641 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5701 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5761 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5821 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5881 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5941 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6001 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6061 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6121 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6181 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6241 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6301 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6361 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6421 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6481 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgct
//
