LOCUS       sequence-269766-        6789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EIF3B (465-552) 3'UTR with the Artificial Unstructured 3'UTR fused 
            upstream 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 6789)
  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 6789)
  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-Unstructured-EIF3B-465-552
FEATURES             Location/Qualifiers
     source          1..6789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        99..456
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        99..290
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(122..142)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      307..442
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          526..658
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        703..721
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      725..734
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             731..1666
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2016..2064
                     /note="synthetic polyadenylation signal"
     promoter        2073..2824
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2894..3026
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        3071..3089
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3098..3107
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3104..4756
                     /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             4063..4071
                     /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    4781..4902
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4792..4811)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4846..4865
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5035..5139
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5140..5208,5209..6000)
                     /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(5358..5377)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6171..6759
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6660..6679
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctggcctttt gctggccttt tgctcacatg gctcgacaga tctgcgcagc accatggcct
       61 gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg
      121 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
      181 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
      241 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
      301 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
      361 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
      421 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttgattcttc tgacacaaca
      481 gtctcgaact taagctgcag aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt
      541 acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg
      601 cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt
      661 gtccactccc agttcaatta cagctcttaa ggctagagta cttaatacga ctcactatag
      721 gctagccacc atggcttcca aggtgtacga ccccgagcaa cgcaaacgca tgatcactgg
      781 gcctcagtgg tgggctcgct gcaagcaaat gaacgtgctg gactccttca tcaactacta
      841 tgattccgag aagcacgccg agaacgccgt gatttttctg catggtaacg ctgcctccag
      901 ctacctgtgg aggcacgtcg tgcctcacat cgagcccgtg gctagatgca tcatccctga
      961 tctgatcgga atgggtaagt ccggcaagag cgggaatggc tcatatcgcc tcctggatca
     1021 ctacaagtac ctcaccgctt ggttcgagct gctgaacctt ccaaagaaaa tcatctttgt
     1081 gggccacgac tggggggctt gtctggcctt tcactactcc tacgagcacc aagacaagat
     1141 caaggccatc gtccatgctg agagtgtcgt ggacgtgatc gagtcctggg acgagtggcc
     1201 tgacatcgag gaggatatcg ccctgatcaa gagcgaagag ggcgagaaaa tggtgcttga
     1261 gaataacttc ttcgtcgaga ccatgctccc aagcaagatc atgcggaaac tggagcctga
     1321 ggagttcgct gcctacctgg agccattcaa ggagaagggc gaggttagac ggcctaccct
     1381 ctcctggcct cgcgagatcc ctctcgttaa gggaggcaag cccgacgtcg tccagattgt
     1441 ccgcaactac aacgcctacc ttcgggccag cgacgatctg cctaagatgt tcatcgagtc
     1501 cgaccctggg ttcttttcca acgctattgt cgagggagct aagaagttcc ctaacaccga
     1561 gttcgtgaag gtgaagggcc tccacttcag ccaggaggac gctccagatg aaatgggtaa
     1621 gtacatcaag agcttcgtgg agcgcgtgct gaagaacgag cagtaattct aggcgatcgc
     1681 tcgagccccc ccctaccctc aacctcaaac ccccaccaaa accacacaac acaaaccatc
     1741 ccacaaacac ccaaaactaa cccccaccaa ccccaaacac catcagagct cactccaccc
     1801 cccccactac acacctaatt tcttcccact atctccaacc actccctaac cataaaaccc
     1861 tccccacaac ctaaccaaac caccaaaact agttaaacgg caccgccatg ccttgtaccc
     1921 ccaccgtgca ggttgtggcc ggttttctcc gcaggttgaa catggaaata aaagcaaact
     1981 tgtatgagtt taaacctaga gcggccgctg gccgcaataa aatatcttta ttttcattac
     2041 atctgtgtgt tggttttttg tgtgaggatc taaatgagtc ttcggacctc gcgggggccg
     2101 cttaagcggt ggttagggtt tgtctgacgc ggggggaggg ggaaggaacg aaacactctc
     2161 attcggaggc ggctcggggt ttggtcttgg tggccacggg cacgcagaag agcgccgcga
     2221 tcctcttaag cacccccccg ccctccgtgg aggcgggggt ttggtcggcg ggtggtaact
     2281 ggcgggccgc tgactcgggc gggtcgcgcg ccccagagtg tgaccttttc ggtctgctcg
     2341 cagacccccg ggcggcgccg ccgcggcggc gacgggctcg ctgggtccta ggctccatgg
     2401 ggaccgtata cgtggacagg ctctggagca tccgcacgac tgcggtgata ttaccggaga
     2461 ccttctgcgg gacgagccgg gtcacgcggc tgacgcggag cgtccgttgg gcgacaaaca
     2521 ccaggacggg gcacaggtac actatcttgt cacccggagg cgcgagggac tgcaggagct
     2581 tcagggagtg gcgcagctgc ttcatccccg tggcccgttg ctcgcgtttg ctggcggtgt
     2641 ccccggaaga aatatatttg catgtcttta gttctatgat gacacaaacc ccgcccagcg
     2701 tcttgtcatt ggcgaattcg aacacgcaga tgcagtcggg gcggcgcggt cccaggtcca
     2761 cttcgcatat taaggtgacg cgtgtggcct cgaacaccga gcgaccctgc agcgacccgc
     2821 ttaaaagctt gattcttctg acacaacagt ctcgaactta agctgcagaa gttggtcgtg
     2881 aggcactggg caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact
     2941 gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga
     3001 catccacttt gcctttctct ccacaggtgt ccactcccag ttcaattaca gctcttaagg
     3061 ctagagtact taatacgact cactataggc tagcaaagcc accatggccg atgctaagaa
     3121 cattaagaag ggccctgctc ccttctaccc tctggaggat ggcaccgctg gcgagcagct
     3181 gcacaaggcc atgaagaggt atgccctggt gcctggcacc attgccttca ccgatgccca
     3241 cattgaggtg gacatcacct atgccgagta cttcgagatg tctgtgcgcc tggccgaggc
     3301 catgaagagg tacggcctga acaccaacca ccgcatcgtg gtgtgctctg agaactctct
     3361 gcagttcttc atgccagtgc tgggcgccct gttcatcgga gtggccgtgg cccctgctaa
     3421 cgacatttac aacgagcgcg agctgctgaa cagcatgggc atttctcagc ctaccgtggt
     3481 gttcgtgtct aagaagggcc tgcagaagat cctgaacgtg cagaagaagc tgcctatcat
     3541 ccagaagatc atcatcatgg actctaagac cgactaccag ggcttccaga gcatgtacac
     3601 attcgtgaca tctcatctgc ctcctggctt caacgagtac gacttcgtgc cagagtcttt
     3661 cgacagggac aaaaccattg ccctgatcat gaacagctct gggtctaccg gcctgcctaa
     3721 gggcgtggcc ctgcctcatc gcaccgcctg tgtgcgcttc tctcacgccc gcgaccctat
     3781 tttcggcaac cagatcatcc ccgacaccgc tattctgagc gtggtgccat tccaccacgg
     3841 cttcggcatg ttcaccaccc tgggctacct gatttgcggc tttcgggtgg tgctgatgta
     3901 ccgcttcgag gaggagctgt tcctgcgcag cctgcaagac tacaaaattc agtctgccct
     3961 gctggtgcca accctgttca gcttcttcgc taagagcacc ctgatcgaca agtacgacct
     4021 gtctaacctg cacgagattg cctctggcgg cgccccactg tctaaggagg tgggcgaagc
     4081 cgtggccaag cgctttcatc tgccaggcat ccgccagggc tacggcctga ccgagacaac
     4141 cagcgccatt ctgattaccc cagagggcga cgacaagcct ggcgccgtgg gcaaggtggt
     4201 gccattcttc gaggccaagg tggtggacct ggacaccggc aagaccctgg gagtgaacca
     4261 gcgcggcgag ctgtgtgtgc gcggccctat gattatgtcc ggctacgtga ataaccctga
     4321 ggccacaaac gccctgatcg acaaggacgg ctggctgcac tctggcgaca ttgcctactg
     4381 ggacgaggac gagcacttct tcatcgtgga ccgcctgaag tctctgatca agtacaaggg
     4441 ctaccaggtg gccccagccg agctggagtc tatcctgctg cagcacccta acattttcga
     4501 cgccggagtg gccggcctgc ccgacgacga tgccggcgag ctgcctgccg ccgtcgtcgt
     4561 gctggaacac ggcaagacca tgaccgagaa ggagatcgtg gactatgtgg ccagccaggt
     4621 gacaaccgcc aagaagctgc gcggcggagt ggtgttcgtg gacgaggtgc ccaagggcct
     4681 gaccggcaag ctggacgccc gcaagatccg cgagatcctg atcaaggcta agaaaggcgg
     4741 caagatcgcc gtgtaatgcg gccgcttcga gcagacatga taagatacat tgatgagttt
     4801 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     4861 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     4921 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     4981 tacaaatgtg gtaaaatcga taaggatcca ggtggcactt ttcggggaaa tgtgcgcgga
     5041 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5101 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5161 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5221 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5281 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5341 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5401 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5461 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5521 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5581 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5641 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5701 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5761 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5821 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5881 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5941 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6001 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6061 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6121 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6181 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6241 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6301 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6361 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6421 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6481 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6541 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6601 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6661 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6721 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6781 ttacggttc
//
