LOCUS       sequence-269946-        6804 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EIF3B (148-464) 3'UTR fragment 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 6804)
  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 6804)
  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-148-464
FEATURES             Location/Qualifiers
     source          1..6804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          109..241
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        286..304
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      308..317
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             314..1249
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1614..1662
                     /note="synthetic polyadenylation signal"
     promoter        1671..2422
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2492..2624
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        2669..2687
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2696..2705
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2702..4354
                     /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             3661..3669
                     /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    4379..4500
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4390..4409)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4444..4463
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4633..4737
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4738..4806,4807..5598)
                     /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(4956..4975)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5769..6357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6258..6277
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(6486..6804,1..39)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6486..6677
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6509..6529)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6694..6804,1..25)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttgattc ttctgacaca
       61 acagtctcga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag
      121 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
      181 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
      241 ggtgtccact cccagttcaa ttacagctct taaggctaga gtacttaata cgactcacta
      301 taggctagcc accatggctt ccaaggtgta cgaccccgag caacgcaaac gcatgatcac
      361 tgggcctcag tggtgggctc gctgcaagca aatgaacgtg ctggactcct tcatcaacta
      421 ctatgattcc gagaagcacg ccgagaacgc cgtgattttt ctgcatggta acgctgcctc
      481 cagctacctg tggaggcacg tcgtgcctca catcgagccc gtggctagat gcatcatccc
      541 tgatctgatc ggaatgggta agtccggcaa gagcgggaat ggctcatatc gcctcctgga
      601 tcactacaag tacctcaccg cttggttcga gctgctgaac cttccaaaga aaatcatctt
      661 tgtgggccac gactgggggg cttgtctggc ctttcactac tcctacgagc accaagacaa
      721 gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg atcgagtcct gggacgagtg
      781 gcctgacatc gaggaggata tcgccctgat caagagcgaa gagggcgaga aaatggtgct
      841 tgagaataac ttcttcgtcg agaccatgct cccaagcaag atcatgcgga aactggagcc
      901 tgaggagttc gctgcctacc tggagccatt caaggagaag ggcgaggtta gacggcctac
      961 cctctcctgg cctcgcgaga tccctctcgt taagggaggc aagcccgacg tcgtccagat
     1021 tgtccgcaac tacaacgcct accttcgggc cagcgacgat ctgcctaaga tgttcatcga
     1081 gtccgaccct gggttctttt ccaacgctat tgtcgaggga gctaagaagt tccctaacac
     1141 cgagttcgtg aaggtgaagg gcctccactt cagccaggag gacgctccag atgaaatggg
     1201 taagtacatc aagagcttcg tggagcgcgt gctgaagaac gagcagtaat tctaggcgat
     1261 cgctcgagct cctccctgtg ctctctggct ctggactgtg actgcgcctg gattctgcca
     1321 ttgcgacaca tttttgtgcc tttcagcccc tggtgtctgc agtgggggat ttaaggcacc
     1381 cgcttccact tctttcttgt ttggagtttt ctgttggaac cgccggcgtt ggctccgaag
     1441 acttagcgac gccactggcg gcaccttctc ctgcgcccag tgatgtttcc acggtgcctg
     1501 tacacagccg agcagcattt ccgttgaagg acttgcatcc ccattgcggg cagtgctgga
     1561 cgtgtcccgg agacccaccg ggagggttta aacctagagc ggccgctggc cgcaataaaa
     1621 tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaggatcta aatgagtctt
     1681 cggacctcgc gggggccgct taagcggtgg ttagggtttg tctgacgcgg ggggaggggg
     1741 aaggaacgaa acactctcat tcggaggcgg ctcggggttt ggtcttggtg gccacgggca
     1801 cgcagaagag cgccgcgatc ctcttaagca cccccccgcc ctccgtggag gcgggggttt
     1861 ggtcggcggg tggtaactgg cgggccgctg actcgggcgg gtcgcgcgcc ccagagtgtg
     1921 accttttcgg tctgctcgca gacccccggg cggcgccgcc gcggcggcga cgggctcgct
     1981 gggtcctagg ctccatgggg accgtatacg tggacaggct ctggagcatc cgcacgactg
     2041 cggtgatatt accggagacc ttctgcggga cgagccgggt cacgcggctg acgcggagcg
     2101 tccgttgggc gacaaacacc aggacggggc acaggtacac tatcttgtca cccggaggcg
     2161 cgagggactg caggagcttc agggagtggc gcagctgctt catccccgtg gcccgttgct
     2221 cgcgtttgct ggcggtgtcc ccggaagaaa tatatttgca tgtctttagt tctatgatga
     2281 cacaaacccc gcccagcgtc ttgtcattgg cgaattcgaa cacgcagatg cagtcggggc
     2341 ggcgcggtcc caggtccact tcgcatatta aggtgacgcg tgtggcctcg aacaccgagc
     2401 gaccctgcag cgacccgctt aaaagcttga ttcttctgac acaacagtct cgaacttaag
     2461 ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta
     2521 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc
     2581 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccagtt
     2641 caattacagc tcttaaggct agagtactta atacgactca ctataggcta gcaaagccac
     2701 catggccgat gctaagaaca ttaagaaggg ccctgctccc ttctaccctc tggaggatgg
     2761 caccgctggc gagcagctgc acaaggccat gaagaggtat gccctggtgc ctggcaccat
     2821 tgccttcacc gatgcccaca ttgaggtgga catcacctat gccgagtact tcgagatgtc
     2881 tgtgcgcctg gccgaggcca tgaagaggta cggcctgaac accaaccacc gcatcgtggt
     2941 gtgctctgag aactctctgc agttcttcat gccagtgctg ggcgccctgt tcatcggagt
     3001 ggccgtggcc cctgctaacg acatttacaa cgagcgcgag ctgctgaaca gcatgggcat
     3061 ttctcagcct accgtggtgt tcgtgtctaa gaagggcctg cagaagatcc tgaacgtgca
     3121 gaagaagctg cctatcatcc agaagatcat catcatggac tctaagaccg actaccaggg
     3181 cttccagagc atgtacacat tcgtgacatc tcatctgcct cctggcttca acgagtacga
     3241 cttcgtgcca gagtctttcg acagggacaa aaccattgcc ctgatcatga acagctctgg
     3301 gtctaccggc ctgcctaagg gcgtggccct gcctcatcgc accgcctgtg tgcgcttctc
     3361 tcacgcccgc gaccctattt tcggcaacca gatcatcccc gacaccgcta ttctgagcgt
     3421 ggtgccattc caccacggct tcggcatgtt caccaccctg ggctacctga tttgcggctt
     3481 tcgggtggtg ctgatgtacc gcttcgagga ggagctgttc ctgcgcagcc tgcaagacta
     3541 caaaattcag tctgccctgc tggtgccaac cctgttcagc ttcttcgcta agagcaccct
     3601 gatcgacaag tacgacctgt ctaacctgca cgagattgcc tctggcggcg ccccactgtc
     3661 taaggaggtg ggcgaagccg tggccaagcg ctttcatctg ccaggcatcc gccagggcta
     3721 cggcctgacc gagacaacca gcgccattct gattacccca gagggcgacg acaagcctgg
     3781 cgccgtgggc aaggtggtgc cattcttcga ggccaaggtg gtggacctgg acaccggcaa
     3841 gaccctggga gtgaaccagc gcggcgagct gtgtgtgcgc ggccctatga ttatgtccgg
     3901 ctacgtgaat aaccctgagg ccacaaacgc cctgatcgac aaggacggct ggctgcactc
     3961 tggcgacatt gcctactggg acgaggacga gcacttcttc atcgtggacc gcctgaagtc
     4021 tctgatcaag tacaagggct accaggtggc cccagccgag ctggagtcta tcctgctgca
     4081 gcaccctaac attttcgacg ccggagtggc cggcctgccc gacgacgatg ccggcgagct
     4141 gcctgccgcc gtcgtcgtgc tggaacacgg caagaccatg accgagaagg agatcgtgga
     4201 ctatgtggcc agccaggtga caaccgccaa gaagctgcgc ggcggagtgg tgttcgtgga
     4261 cgaggtgccc aagggcctga ccggcaagct ggacgcccgc aagatccgcg agatcctgat
     4321 caaggctaag aaaggcggca agatcgccgt gtaatgcggc cgcttcgagc agacatgata
     4381 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     4441 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     4501 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     4561 taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccagg tggcactttt
     4621 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4681 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4741 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4801 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4861 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4921 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4981 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5041 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5101 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5161 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5221 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5281 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5341 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5401 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5461 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5521 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5581 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5641 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5701 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5761 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5821 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5881 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5941 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6001 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6061 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6121 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6181 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6241 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6301 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6361 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatggct
     6421 cgacagatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt
     6481 accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     6541 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     6601 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     6661 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     6721 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     6781 tcggcctctg agctattcca gaag
//
