LOCUS       sequence-269955-        7253 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EIF3B 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 7253)
  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 7253)
  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
FEATURES             Location/Qualifiers
     source          1..7253
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          178..310
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        355..373
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      377..386
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             383..1318
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2132..2180
                     /note="synthetic polyadenylation signal"
     promoter        2189..2940
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          3010..3142
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        3187..3205
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3214..3223
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3220..4872
                     /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             4179..4187
                     /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    4897..5018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4908..4927)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4962..4981
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5151..5255
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5256..5324,5325..6116)
                     /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(5474..5493)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6287..6875
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6776..6795
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(7004..7253,1..108)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        7004..7195
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(7027..7047)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(7212..7253,1..94)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
       61 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttgattct
      121 tctgacacaa cagtctcgaa cttaagctgc agaagttggt cgtgaggcac tgggcaggta
      181 agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag
      241 agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt
      301 ctctccacag gtgtccactc ccagttcaat tacagctctt aaggctagag tacttaatac
      361 gactcactat aggctagcca ccatggcttc caaggtgtac gaccccgagc aacgcaaacg
      421 catgatcact gggcctcagt ggtgggctcg ctgcaagcaa atgaacgtgc tggactcctt
      481 catcaactac tatgattccg agaagcacgc cgagaacgcc gtgatttttc tgcatggtaa
      541 cgctgcctcc agctacctgt ggaggcacgt cgtgcctcac atcgagcccg tggctagatg
      601 catcatccct gatctgatcg gaatgggtaa gtccggcaag agcgggaatg gctcatatcg
      661 cctcctggat cactacaagt acctcaccgc ttggttcgag ctgctgaacc ttccaaagaa
      721 aatcatcttt gtgggccacg actggggggc ttgtctggcc tttcactact cctacgagca
      781 ccaagacaag atcaaggcca tcgtccatgc tgagagtgtc gtggacgtga tcgagtcctg
      841 ggacgagtgg cctgacatcg aggaggatat cgccctgatc aagagcgaag agggcgagaa
      901 aatggtgctt gagaataact tcttcgtcga gaccatgctc ccaagcaaga tcatgcggaa
      961 actggagcct gaggagttcg ctgcctacct ggagccattc aaggagaagg gcgaggttag
     1021 acggcctacc ctctcctggc ctcgcgagat ccctctcgtt aagggaggca agcccgacgt
     1081 cgtccagatt gtccgcaact acaacgccta ccttcgggcc agcgacgatc tgcctaagat
     1141 gttcatcgag tccgaccctg ggttcttttc caacgctatt gtcgagggag ctaagaagtt
     1201 ccctaacacc gagttcgtga aggtgaaggg cctccacttc agccaggagg acgctccaga
     1261 tgaaatgggt aagtacatca agagcttcgt ggagcgcgtg ctgaagaacg agcagtaatt
     1321 ctaggcgatc gctcgagccc ccccctaccc tcaacctcaa acccccacca aaaccacaca
     1381 acacaaacca tcccacaaac acccaaaact aacccccacc aaccccaaac accatcagag
     1441 ctcactccac cccccccact acacacctaa tttcttccca ctatctccaa ccactcccta
     1501 accataaaac cctccccaca acctaaccaa accaccaaaa ctagtttaaa ccctggagca
     1561 ctgtggggac ggactccgcc tgctgttccc gcgctgagct acaggactcc cgagtgtgag
     1621 ccgcggttcc tctgttgcag cgcagccgtg tgtgctgtgg agccgaggcc gtcctgcagg
     1681 aagccgcgtg actcccgact cctccctgtg ctctctggct ctggactgtg actgcgcctg
     1741 gattctgcca ttgcgacaca tttttgtgcc tttcagcccc tggtgtctgc agtgggggat
     1801 ttaaggcacc cgcttccact tctttcttgt ttggagtttt ctgttggaac cgccggcgtt
     1861 ggctccgaag acttagcgac gccactggcg gcaccttctc ctgcgcccag tgatgtttcc
     1921 acggtgcctg tacacagccg agcagcattt ccgttgaagg acttgcatcc ccattgcggg
     1981 cagtgctgga cgtgtcccgg agacccaccg ggagggcacc gccatgcctt gtacccccac
     2041 cgtgcaggtt gtggccggtt ttctccgcag gttgaacatg gaaataaaag caaacttgta
     2101 tgagtttaaa cctagagcgg ccgctggccg caataaaata tctttatttt cattacatct
     2161 gtgtgttggt tttttgtgtg aggatctaaa tgagtcttcg gacctcgcgg gggccgctta
     2221 agcggtggtt agggtttgtc tgacgcgggg ggagggggaa ggaacgaaac actctcattc
     2281 ggaggcggct cggggtttgg tcttggtggc cacgggcacg cagaagagcg ccgcgatcct
     2341 cttaagcacc cccccgccct ccgtggaggc gggggtttgg tcggcgggtg gtaactggcg
     2401 ggccgctgac tcgggcgggt cgcgcgcccc agagtgtgac cttttcggtc tgctcgcaga
     2461 cccccgggcg gcgccgccgc ggcggcgacg ggctcgctgg gtcctaggct ccatggggac
     2521 cgtatacgtg gacaggctct ggagcatccg cacgactgcg gtgatattac cggagacctt
     2581 ctgcgggacg agccgggtca cgcggctgac gcggagcgtc cgttgggcga caaacaccag
     2641 gacggggcac aggtacacta tcttgtcacc cggaggcgcg agggactgca ggagcttcag
     2701 ggagtggcgc agctgcttca tccccgtggc ccgttgctcg cgtttgctgg cggtgtcccc
     2761 ggaagaaata tatttgcatg tctttagttc tatgatgaca caaaccccgc ccagcgtctt
     2821 gtcattggcg aattcgaaca cgcagatgca gtcggggcgg cgcggtccca ggtccacttc
     2881 gcatattaag gtgacgcgtg tggcctcgaa caccgagcga ccctgcagcg acccgcttaa
     2941 aagcttgatt cttctgacac aacagtctcg aacttaagct gcagaagttg gtcgtgaggc
     3001 actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc
     3061 ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc
     3121 cactttgcct ttctctccac aggtgtccac tcccagttca attacagctc ttaaggctag
     3181 agtacttaat acgactcact ataggctagc aaagccacca tggccgatgc taagaacatt
     3241 aagaagggcc ctgctccctt ctaccctctg gaggatggca ccgctggcga gcagctgcac
     3301 aaggccatga agaggtatgc cctggtgcct ggcaccattg ccttcaccga tgcccacatt
     3361 gaggtggaca tcacctatgc cgagtacttc gagatgtctg tgcgcctggc cgaggccatg
     3421 aagaggtacg gcctgaacac caaccaccgc atcgtggtgt gctctgagaa ctctctgcag
     3481 ttcttcatgc cagtgctggg cgccctgttc atcggagtgg ccgtggcccc tgctaacgac
     3541 atttacaacg agcgcgagct gctgaacagc atgggcattt ctcagcctac cgtggtgttc
     3601 gtgtctaaga agggcctgca gaagatcctg aacgtgcaga agaagctgcc tatcatccag
     3661 aagatcatca tcatggactc taagaccgac taccagggct tccagagcat gtacacattc
     3721 gtgacatctc atctgcctcc tggcttcaac gagtacgact tcgtgccaga gtctttcgac
     3781 agggacaaaa ccattgccct gatcatgaac agctctgggt ctaccggcct gcctaagggc
     3841 gtggccctgc ctcatcgcac cgcctgtgtg cgcttctctc acgcccgcga ccctattttc
     3901 ggcaaccaga tcatccccga caccgctatt ctgagcgtgg tgccattcca ccacggcttc
     3961 ggcatgttca ccaccctggg ctacctgatt tgcggctttc gggtggtgct gatgtaccgc
     4021 ttcgaggagg agctgttcct gcgcagcctg caagactaca aaattcagtc tgccctgctg
     4081 gtgccaaccc tgttcagctt cttcgctaag agcaccctga tcgacaagta cgacctgtct
     4141 aacctgcacg agattgcctc tggcggcgcc ccactgtcta aggaggtggg cgaagccgtg
     4201 gccaagcgct ttcatctgcc aggcatccgc cagggctacg gcctgaccga gacaaccagc
     4261 gccattctga ttaccccaga gggcgacgac aagcctggcg ccgtgggcaa ggtggtgcca
     4321 ttcttcgagg ccaaggtggt ggacctggac accggcaaga ccctgggagt gaaccagcgc
     4381 ggcgagctgt gtgtgcgcgg ccctatgatt atgtccggct acgtgaataa ccctgaggcc
     4441 acaaacgccc tgatcgacaa ggacggctgg ctgcactctg gcgacattgc ctactgggac
     4501 gaggacgagc acttcttcat cgtggaccgc ctgaagtctc tgatcaagta caagggctac
     4561 caggtggccc cagccgagct ggagtctatc ctgctgcagc accctaacat tttcgacgcc
     4621 ggagtggccg gcctgcccga cgacgatgcc ggcgagctgc ctgccgccgt cgtcgtgctg
     4681 gaacacggca agaccatgac cgagaaggag atcgtggact atgtggccag ccaggtgaca
     4741 accgccaaga agctgcgcgg cggagtggtg ttcgtggacg aggtgcccaa gggcctgacc
     4801 ggcaagctgg acgcccgcaa gatccgcgag atcctgatca aggctaagaa aggcggcaag
     4861 atcgccgtgt aatgcggccg cttcgagcag acatgataag atacattgat gagtttggac
     4921 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     4981 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     5041 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     5101 aatgtggtaa aatcgataag gatccaggtg gcacttttcg gggaaatgtg cgcggaaccc
     5161 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     5221 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     5281 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     5341 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     5401 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     5461 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     5521 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     5581 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     5641 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     5701 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     5761 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     5821 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     5881 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5941 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     6001 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     6061 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     6121 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     6181 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6241 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6301 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6361 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6421 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6481 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6541 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6601 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6661 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6721 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6781 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6841 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6901 ggttcctggc cttttgctgg ccttttgctc acatggctcg acagatctgc gcagcaccat
     6961 ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc
     7021 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     7081 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     7141 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     7201 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccc
//
