LOCUS       sequence-269765-        6533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EIF3B (465-552) 3'UTR with the main hairpin deleted 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 6533)
  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 6533)
  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-Delta-Hairpin
FEATURES             Location/Qualifiers
     source          1..6533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          78..210
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        255..273
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      277..286
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             283..1218
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1312..1360
                     /note="synthetic polyadenylation signal"
     promoter        1369..2120
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2190..2322
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        2367..2385
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2394..2403
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2400..4052
                     /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             3359..3367
                     /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    4077..4198
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4088..4107)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4142..4161
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4331..4435
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4436..4504,4505..5296)
                     /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(4654..4673)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5467..6055
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5956..5975
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(6184..6533,1..8)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6184..6375
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6207..6227)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6392..6527
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa cttaagctgc agaagttggt
       61 cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga
      121 aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta
      181 ctgacatcca ctttgccttt ctctccacag gtgtccactc ccagttcaat tacagctctt
      241 aaggctagag tacttaatac gactcactat aggctagcca ccatggcttc caaggtgtac
      301 gaccccgagc aacgcaaacg catgatcact gggcctcagt ggtgggctcg ctgcaagcaa
      361 atgaacgtgc tggactcctt catcaactac tatgattccg agaagcacgc cgagaacgcc
      421 gtgatttttc tgcatggtaa cgctgcctcc agctacctgt ggaggcacgt cgtgcctcac
      481 atcgagcccg tggctagatg catcatccct gatctgatcg gaatgggtaa gtccggcaag
      541 agcgggaatg gctcatatcg cctcctggat cactacaagt acctcaccgc ttggttcgag
      601 ctgctgaacc ttccaaagaa aatcatcttt gtgggccacg actggggggc ttgtctggcc
      661 tttcactact cctacgagca ccaagacaag atcaaggcca tcgtccatgc tgagagtgtc
      721 gtggacgtga tcgagtcctg ggacgagtgg cctgacatcg aggaggatat cgccctgatc
      781 aagagcgaag agggcgagaa aatggtgctt gagaataact tcttcgtcga gaccatgctc
      841 ccaagcaaga tcatgcggaa actggagcct gaggagttcg ctgcctacct ggagccattc
      901 aaggagaagg gcgaggttag acggcctacc ctctcctggc ctcgcgagat ccctctcgtt
      961 aagggaggca agcccgacgt cgtccagatt gtccgcaact acaacgccta ccttcgggcc
     1021 agcgacgatc tgcctaagat gttcatcgag tccgaccctg ggttcttttc caacgctatt
     1081 gtcgagggag ctaagaagtt ccctaacacc gagttcgtga aggtgaaggg cctccacttc
     1141 agccaggagg acgctccaga tgaaatgggt aagtacatca agagcttcgt ggagcgcgtg
     1201 ctgaagaacg agcagtaatt ctaggcgatc gctcgaggct ttctccgcag gttgaacatg
     1261 gaaataaaag caaacttgta tgagtttaaa cctagagcgg ccgctggccg caataaaata
     1321 tctttatttt cattacatct gtgtgttggt tttttgtgtg aggatctaaa tgagtcttcg
     1381 gacctcgcgg gggccgctta agcggtggtt agggtttgtc tgacgcgggg ggagggggaa
     1441 ggaacgaaac actctcattc ggaggcggct cggggtttgg tcttggtggc cacgggcacg
     1501 cagaagagcg ccgcgatcct cttaagcacc cccccgccct ccgtggaggc gggggtttgg
     1561 tcggcgggtg gtaactggcg ggccgctgac tcgggcgggt cgcgcgcccc agagtgtgac
     1621 cttttcggtc tgctcgcaga cccccgggcg gcgccgccgc ggcggcgacg ggctcgctgg
     1681 gtcctaggct ccatggggac cgtatacgtg gacaggctct ggagcatccg cacgactgcg
     1741 gtgatattac cggagacctt ctgcgggacg agccgggtca cgcggctgac gcggagcgtc
     1801 cgttgggcga caaacaccag gacggggcac aggtacacta tcttgtcacc cggaggcgcg
     1861 agggactgca ggagcttcag ggagtggcgc agctgcttca tccccgtggc ccgttgctcg
     1921 cgtttgctgg cggtgtcccc ggaagaaata tatttgcatg tctttagttc tatgatgaca
     1981 caaaccccgc ccagcgtctt gtcattggcg aattcgaaca cgcagatgca gtcggggcgg
     2041 cgcggtccca ggtccacttc gcatattaag gtgacgcgtg tggcctcgaa caccgagcga
     2101 ccctgcagcg acccgcttaa aagcttgatt cttctgacac aacagtctcg aacttaagct
     2161 gcagaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag
     2221 gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac
     2281 ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca
     2341 attacagctc ttaaggctag agtacttaat acgactcact ataggctagc aaagccacca
     2401 tggccgatgc taagaacatt aagaagggcc ctgctccctt ctaccctctg gaggatggca
     2461 ccgctggcga gcagctgcac aaggccatga agaggtatgc cctggtgcct ggcaccattg
     2521 ccttcaccga tgcccacatt gaggtggaca tcacctatgc cgagtacttc gagatgtctg
     2581 tgcgcctggc cgaggccatg aagaggtacg gcctgaacac caaccaccgc atcgtggtgt
     2641 gctctgagaa ctctctgcag ttcttcatgc cagtgctggg cgccctgttc atcggagtgg
     2701 ccgtggcccc tgctaacgac atttacaacg agcgcgagct gctgaacagc atgggcattt
     2761 ctcagcctac cgtggtgttc gtgtctaaga agggcctgca gaagatcctg aacgtgcaga
     2821 agaagctgcc tatcatccag aagatcatca tcatggactc taagaccgac taccagggct
     2881 tccagagcat gtacacattc gtgacatctc atctgcctcc tggcttcaac gagtacgact
     2941 tcgtgccaga gtctttcgac agggacaaaa ccattgccct gatcatgaac agctctgggt
     3001 ctaccggcct gcctaagggc gtggccctgc ctcatcgcac cgcctgtgtg cgcttctctc
     3061 acgcccgcga ccctattttc ggcaaccaga tcatccccga caccgctatt ctgagcgtgg
     3121 tgccattcca ccacggcttc ggcatgttca ccaccctggg ctacctgatt tgcggctttc
     3181 gggtggtgct gatgtaccgc ttcgaggagg agctgttcct gcgcagcctg caagactaca
     3241 aaattcagtc tgccctgctg gtgccaaccc tgttcagctt cttcgctaag agcaccctga
     3301 tcgacaagta cgacctgtct aacctgcacg agattgcctc tggcggcgcc ccactgtcta
     3361 aggaggtggg cgaagccgtg gccaagcgct ttcatctgcc aggcatccgc cagggctacg
     3421 gcctgaccga gacaaccagc gccattctga ttaccccaga gggcgacgac aagcctggcg
     3481 ccgtgggcaa ggtggtgcca ttcttcgagg ccaaggtggt ggacctggac accggcaaga
     3541 ccctgggagt gaaccagcgc ggcgagctgt gtgtgcgcgg ccctatgatt atgtccggct
     3601 acgtgaataa ccctgaggcc acaaacgccc tgatcgacaa ggacggctgg ctgcactctg
     3661 gcgacattgc ctactgggac gaggacgagc acttcttcat cgtggaccgc ctgaagtctc
     3721 tgatcaagta caagggctac caggtggccc cagccgagct ggagtctatc ctgctgcagc
     3781 accctaacat tttcgacgcc ggagtggccg gcctgcccga cgacgatgcc ggcgagctgc
     3841 ctgccgccgt cgtcgtgctg gaacacggca agaccatgac cgagaaggag atcgtggact
     3901 atgtggccag ccaggtgaca accgccaaga agctgcgcgg cggagtggtg ttcgtggacg
     3961 aggtgcccaa gggcctgacc ggcaagctgg acgcccgcaa gatccgcgag atcctgatca
     4021 aggctaagaa aggcggcaag atcgccgtgt aatgcggccg cttcgagcag acatgataag
     4081 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     4141 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     4201 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     4261 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccaggtg gcacttttcg
     4321 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4381 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4441 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4501 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4561 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4621 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4681 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4741 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4801 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4861 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4921 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4981 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5041 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5101 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5161 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5221 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5281 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5341 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5401 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5461 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5521 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5581 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5641 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5701 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5761 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5821 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5881 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5941 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6001 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6061 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatggctcg
     6121 acagatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac
     6181 cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     6241 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     6301 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     6361 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     6421 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     6481 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gct
//
