LOCUS       sequence-269763-        6575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EIF3B (465-552) 3'UTR with the main hairpin mutated 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-88nt-Disrupted-Hairpin
FEATURES             Location/Qualifiers
     source          1..6575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6..141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          225..357
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        402..420
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      424..433
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             430..1365
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1501..1549
                     /note="synthetic polyadenylation signal"
     promoter        1558..2309
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2379..2511
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        2556..2574
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2583..2592
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2589..4241
                     /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             3548..3556
                     /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    4266..4387
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4277..4296)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4331..4350
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4520..4624
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4625..4693,4694..5485)
                     /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(4843..4862)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5656..6244
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6145..6164
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(6373..6575,1..155)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6373..6564
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6396..6416)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
       61 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
      121 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tgattcttct gacacaacag
      181 tctcgaactt aagctgcaga agttggtcgt gaggcactgg gcaggtaagt atcaaggtta
      241 caagacaggt ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc
      301 gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg
      361 tccactccca gttcaattac agctcttaag gctagagtac ttaatacgac tcactatagg
      421 ctagccacca tggcttccaa ggtgtacgac cccgagcaac gcaaacgcat gatcactggg
      481 cctcagtggt gggctcgctg caagcaaatg aacgtgctgg actccttcat caactactat
      541 gattccgaga agcacgccga gaacgccgtg atttttctgc atggtaacgc tgcctccagc
      601 tacctgtgga ggcacgtcgt gcctcacatc gagcccgtgg ctagatgcat catccctgat
      661 ctgatcggaa tgggtaagtc cggcaagagc gggaatggct catatcgcct cctggatcac
      721 tacaagtacc tcaccgcttg gttcgagctg ctgaaccttc caaagaaaat catctttgtg
      781 ggccacgact ggggggcttg tctggccttt cactactcct acgagcacca agacaagatc
      841 aaggccatcg tccatgctga gagtgtcgtg gacgtgatcg agtcctggga cgagtggcct
      901 gacatcgagg aggatatcgc cctgatcaag agcgaagagg gcgagaaaat ggtgcttgag
      961 aataacttct tcgtcgagac catgctccca agcaagatca tgcggaaact ggagcctgag
     1021 gagttcgctg cctacctgga gccattcaag gagaagggcg aggttagacg gcctaccctc
     1081 tcctggcctc gcgagatccc tctcgttaag ggaggcaagc ccgacgtcgt ccagattgtc
     1141 cgcaactaca acgcctacct tcgggccagc gacgatctgc ctaagatgtt catcgagtcc
     1201 gaccctgggt tcttttccaa cgctattgtc gagggagcta agaagttccc taacaccgag
     1261 ttcgtgaagg tgaagggcct ccacttcagc caggaggacg ctccagatga aatgggtaag
     1321 tacatcaaga gcttcgtgga gcgcgtgctg aagaacgagc agtaattcta ggcgatcgct
     1381 cgaggctggt ggtacccttg tacccccacc gtgcaggttg tggccggttt tctccgcagg
     1441 ttgaacatgg aaataaaagc aaacttgtat gagtttaaac ctagagcggc cgctggccgc
     1501 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga ggatctaaat
     1561 gagtcttcgg acctcgcggg ggccgcttaa gcggtggtta gggtttgtct gacgcggggg
     1621 gagggggaag gaacgaaaca ctctcattcg gaggcggctc ggggtttggt cttggtggcc
     1681 acgggcacgc agaagagcgc cgcgatcctc ttaagcaccc ccccgccctc cgtggaggcg
     1741 ggggtttggt cggcgggtgg taactggcgg gccgctgact cgggcgggtc gcgcgcccca
     1801 gagtgtgacc ttttcggtct gctcgcagac ccccgggcgg cgccgccgcg gcggcgacgg
     1861 gctcgctggg tcctaggctc catggggacc gtatacgtgg acaggctctg gagcatccgc
     1921 acgactgcgg tgatattacc ggagaccttc tgcgggacga gccgggtcac gcggctgacg
     1981 cggagcgtcc gttgggcgac aaacaccagg acggggcaca ggtacactat cttgtcaccc
     2041 ggaggcgcga gggactgcag gagcttcagg gagtggcgca gctgcttcat ccccgtggcc
     2101 cgttgctcgc gtttgctggc ggtgtccccg gaagaaatat atttgcatgt ctttagttct
     2161 atgatgacac aaaccccgcc cagcgtcttg tcattggcga attcgaacac gcagatgcag
     2221 tcggggcggc gcggtcccag gtccacttcg catattaagg tgacgcgtgt ggcctcgaac
     2281 accgagcgac cctgcagcga cccgcttaaa agcttgattc ttctgacaca acagtctcga
     2341 acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac
     2401 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
     2461 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact
     2521 cccagttcaa ttacagctct taaggctaga gtacttaata cgactcacta taggctagca
     2581 aagccaccat ggccgatgct aagaacatta agaagggccc tgctcccttc taccctctgg
     2641 aggatggcac cgctggcgag cagctgcaca aggccatgaa gaggtatgcc ctggtgcctg
     2701 gcaccattgc cttcaccgat gcccacattg aggtggacat cacctatgcc gagtacttcg
     2761 agatgtctgt gcgcctggcc gaggccatga agaggtacgg cctgaacacc aaccaccgca
     2821 tcgtggtgtg ctctgagaac tctctgcagt tcttcatgcc agtgctgggc gccctgttca
     2881 tcggagtggc cgtggcccct gctaacgaca tttacaacga gcgcgagctg ctgaacagca
     2941 tgggcatttc tcagcctacc gtggtgttcg tgtctaagaa gggcctgcag aagatcctga
     3001 acgtgcagaa gaagctgcct atcatccaga agatcatcat catggactct aagaccgact
     3061 accagggctt ccagagcatg tacacattcg tgacatctca tctgcctcct ggcttcaacg
     3121 agtacgactt cgtgccagag tctttcgaca gggacaaaac cattgccctg atcatgaaca
     3181 gctctgggtc taccggcctg cctaagggcg tggccctgcc tcatcgcacc gcctgtgtgc
     3241 gcttctctca cgcccgcgac cctattttcg gcaaccagat catccccgac accgctattc
     3301 tgagcgtggt gccattccac cacggcttcg gcatgttcac caccctgggc tacctgattt
     3361 gcggctttcg ggtggtgctg atgtaccgct tcgaggagga gctgttcctg cgcagcctgc
     3421 aagactacaa aattcagtct gccctgctgg tgccaaccct gttcagcttc ttcgctaaga
     3481 gcaccctgat cgacaagtac gacctgtcta acctgcacga gattgcctct ggcggcgccc
     3541 cactgtctaa ggaggtgggc gaagccgtgg ccaagcgctt tcatctgcca ggcatccgcc
     3601 agggctacgg cctgaccgag acaaccagcg ccattctgat taccccagag ggcgacgaca
     3661 agcctggcgc cgtgggcaag gtggtgccat tcttcgaggc caaggtggtg gacctggaca
     3721 ccggcaagac cctgggagtg aaccagcgcg gcgagctgtg tgtgcgcggc cctatgatta
     3781 tgtccggcta cgtgaataac cctgaggcca caaacgccct gatcgacaag gacggctggc
     3841 tgcactctgg cgacattgcc tactgggacg aggacgagca cttcttcatc gtggaccgcc
     3901 tgaagtctct gatcaagtac aagggctacc aggtggcccc agccgagctg gagtctatcc
     3961 tgctgcagca ccctaacatt ttcgacgccg gagtggccgg cctgcccgac gacgatgccg
     4021 gcgagctgcc tgccgccgtc gtcgtgctgg aacacggcaa gaccatgacc gagaaggaga
     4081 tcgtggacta tgtggccagc caggtgacaa ccgccaagaa gctgcgcggc ggagtggtgt
     4141 tcgtggacga ggtgcccaag ggcctgaccg gcaagctgga cgcccgcaag atccgcgaga
     4201 tcctgatcaa ggctaagaaa ggcggcaaga tcgccgtgta atgcggccgc ttcgagcaga
     4261 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     4321 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     4381 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     4441 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccaggtgg
     4501 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4561 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4621 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4681 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4741 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4801 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4861 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4921 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4981 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5041 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5101 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5161 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5221 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5281 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5341 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5401 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5461 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5521 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5581 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5641 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5701 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5761 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5821 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5881 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5941 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6001 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6061 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6121 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6181 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6241 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6301 catggctcga cagatctgcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg
     6361 gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     6421 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     6481 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6541 caattagtca gcaaccatag tcccgcccct aactc
//
