LOCUS       sequence-269661-        6575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EIF3B (465-552) 3'UTR with the main hairpin mutated to A and U 
            nucleotides only 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-A:U-Hairpin
FEATURES             Location/Qualifiers
     source          1..6575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          97..229
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        274..292
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      296..305
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             302..1237
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1373..1421
                     /note="synthetic polyadenylation signal"
     promoter        1430..2181
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2251..2383
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        2428..2446
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2455..2464
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2461..4113
                     /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             3420..3428
                     /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    4138..4259
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4149..4168)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4203..4222
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4392..4496
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4497..4565,4566..5357)
                     /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(4715..4734)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5528..6116
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6017..6036
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(6245..6575,1..27)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6245..6436
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6268..6288)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6453..6575,1..13)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ttttttggag gcctaggctt ttgcaaaaag cttgattctt ctgacacaac agtctcgaac
       61 ttaagctgca gaagttggtc gtgaggcact gggcaggtaa gtatcaaggt tacaagacag
      121 gtttaaggag accaatagaa actgggcttg tcgagacaga gaagactctt gcgtttctga
      181 taggcaccta ttggtcttac tgacatccac tttgcctttc tctccacagg tgtccactcc
      241 cagttcaatt acagctctta aggctagagt acttaatacg actcactata ggctagccac
      301 catggcttcc aaggtgtacg accccgagca acgcaaacgc atgatcactg ggcctcagtg
      361 gtgggctcgc tgcaagcaaa tgaacgtgct ggactccttc atcaactact atgattccga
      421 gaagcacgcc gagaacgccg tgatttttct gcatggtaac gctgcctcca gctacctgtg
      481 gaggcacgtc gtgcctcaca tcgagcccgt ggctagatgc atcatccctg atctgatcgg
      541 aatgggtaag tccggcaaga gcgggaatgg ctcatatcgc ctcctggatc actacaagta
      601 cctcaccgct tggttcgagc tgctgaacct tccaaagaaa atcatctttg tgggccacga
      661 ctggggggct tgtctggcct ttcactactc ctacgagcac caagacaaga tcaaggccat
      721 cgtccatgct gagagtgtcg tggacgtgat cgagtcctgg gacgagtggc ctgacatcga
      781 ggaggatatc gccctgatca agagcgaaga gggcgagaaa atggtgcttg agaataactt
      841 cttcgtcgag accatgctcc caagcaagat catgcggaaa ctggagcctg aggagttcgc
      901 tgcctacctg gagccattca aggagaaggg cgaggttaga cggcctaccc tctcctggcc
      961 tcgcgagatc cctctcgtta agggaggcaa gcccgacgtc gtccagattg tccgcaacta
     1021 caacgcctac cttcgggcca gcgacgatct gcctaagatg ttcatcgagt ccgaccctgg
     1081 gttcttttcc aacgctattg tcgagggagc taagaagttc cctaacaccg agttcgtgaa
     1141 ggtgaagggc ctccacttca gccaggagga cgctccagat gaaatgggta agtacatcaa
     1201 gagcttcgtg gagcgcgtgc tgaagaacga gcagtaattc taggcgatcg ctcgaggcat
     1261 tattatacct tgtaccccca ccgtgcaggt tataattaat tttctccgca ggttgaacat
     1321 ggaaataaaa gcaaacttgt atgagtttaa acctagagcg gccgctggcc gcaataaaat
     1381 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaggatctaa atgagtcttc
     1441 ggacctcgcg ggggccgctt aagcggtggt tagggtttgt ctgacgcggg gggaggggga
     1501 aggaacgaaa cactctcatt cggaggcggc tcggggtttg gtcttggtgg ccacgggcac
     1561 gcagaagagc gccgcgatcc tcttaagcac ccccccgccc tccgtggagg cgggggtttg
     1621 gtcggcgggt ggtaactggc gggccgctga ctcgggcggg tcgcgcgccc cagagtgtga
     1681 ccttttcggt ctgctcgcag acccccgggc ggcgccgccg cggcggcgac gggctcgctg
     1741 ggtcctaggc tccatgggga ccgtatacgt ggacaggctc tggagcatcc gcacgactgc
     1801 ggtgatatta ccggagacct tctgcgggac gagccgggtc acgcggctga cgcggagcgt
     1861 ccgttgggcg acaaacacca ggacggggca caggtacact atcttgtcac ccggaggcgc
     1921 gagggactgc aggagcttca gggagtggcg cagctgcttc atccccgtgg cccgttgctc
     1981 gcgtttgctg gcggtgtccc cggaagaaat atatttgcat gtctttagtt ctatgatgac
     2041 acaaaccccg cccagcgtct tgtcattggc gaattcgaac acgcagatgc agtcggggcg
     2101 gcgcggtccc aggtccactt cgcatattaa ggtgacgcgt gtggcctcga acaccgagcg
     2161 accctgcagc gacccgctta aaagcttgat tcttctgaca caacagtctc gaacttaagc
     2221 tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa
     2281 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
     2341 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc
     2401 aattacagct cttaaggcta gagtacttaa tacgactcac tataggctag caaagccacc
     2461 atggccgatg ctaagaacat taagaagggc cctgctccct tctaccctct ggaggatggc
     2521 accgctggcg agcagctgca caaggccatg aagaggtatg ccctggtgcc tggcaccatt
     2581 gccttcaccg atgcccacat tgaggtggac atcacctatg ccgagtactt cgagatgtct
     2641 gtgcgcctgg ccgaggccat gaagaggtac ggcctgaaca ccaaccaccg catcgtggtg
     2701 tgctctgaga actctctgca gttcttcatg ccagtgctgg gcgccctgtt catcggagtg
     2761 gccgtggccc ctgctaacga catttacaac gagcgcgagc tgctgaacag catgggcatt
     2821 tctcagccta ccgtggtgtt cgtgtctaag aagggcctgc agaagatcct gaacgtgcag
     2881 aagaagctgc ctatcatcca gaagatcatc atcatggact ctaagaccga ctaccagggc
     2941 ttccagagca tgtacacatt cgtgacatct catctgcctc ctggcttcaa cgagtacgac
     3001 ttcgtgccag agtctttcga cagggacaaa accattgccc tgatcatgaa cagctctggg
     3061 tctaccggcc tgcctaaggg cgtggccctg cctcatcgca ccgcctgtgt gcgcttctct
     3121 cacgcccgcg accctatttt cggcaaccag atcatccccg acaccgctat tctgagcgtg
     3181 gtgccattcc accacggctt cggcatgttc accaccctgg gctacctgat ttgcggcttt
     3241 cgggtggtgc tgatgtaccg cttcgaggag gagctgttcc tgcgcagcct gcaagactac
     3301 aaaattcagt ctgccctgct ggtgccaacc ctgttcagct tcttcgctaa gagcaccctg
     3361 atcgacaagt acgacctgtc taacctgcac gagattgcct ctggcggcgc cccactgtct
     3421 aaggaggtgg gcgaagccgt ggccaagcgc tttcatctgc caggcatccg ccagggctac
     3481 ggcctgaccg agacaaccag cgccattctg attaccccag agggcgacga caagcctggc
     3541 gccgtgggca aggtggtgcc attcttcgag gccaaggtgg tggacctgga caccggcaag
     3601 accctgggag tgaaccagcg cggcgagctg tgtgtgcgcg gccctatgat tatgtccggc
     3661 tacgtgaata accctgaggc cacaaacgcc ctgatcgaca aggacggctg gctgcactct
     3721 ggcgacattg cctactggga cgaggacgag cacttcttca tcgtggaccg cctgaagtct
     3781 ctgatcaagt acaagggcta ccaggtggcc ccagccgagc tggagtctat cctgctgcag
     3841 caccctaaca ttttcgacgc cggagtggcc ggcctgcccg acgacgatgc cggcgagctg
     3901 cctgccgccg tcgtcgtgct ggaacacggc aagaccatga ccgagaagga gatcgtggac
     3961 tatgtggcca gccaggtgac aaccgccaag aagctgcgcg gcggagtggt gttcgtggac
     4021 gaggtgccca agggcctgac cggcaagctg gacgcccgca agatccgcga gatcctgatc
     4081 aaggctaaga aaggcggcaa gatcgccgtg taatgcggcc gcttcgagca gacatgataa
     4141 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     4201 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     4261 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     4321 aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatccaggt ggcacttttc
     4381 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4441 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4501 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4561 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4621 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4681 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4741 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4801 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4861 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4921 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4981 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5041 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5101 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5161 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5221 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5281 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5341 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     5401 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5461 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5521 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5581 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5641 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5701 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5761 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5821 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5881 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5941 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6001 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6061 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6121 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatggctc
     6181 gacagatctg cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta
     6241 ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     6301 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     6361 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     6421 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     6481 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     6541 cggcctctga gctattccag aagtagtgag gaggc
//
