LOCUS       sequence-270191-        7251 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EIF3B 3'UTR with the Artificial Unstructured 3'UTR fused downstream 
            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 7251)
  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 7251)
  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-Unstructured
FEATURES             Location/Qualifiers
     source          1..7251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        13..370
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        13..204
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(36..56)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      221..356
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          440..572
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        617..635
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      639..648
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             645..1580
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2392..2440
                     /note="synthetic polyadenylation signal"
     promoter        2449..3200
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          3270..3402
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        3447..3465
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3474..3483
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3480..5132
                     /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             4439..4447
                     /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    5157..5278
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5168..5187)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5222..5241
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5411..5515
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5516..5584,5585..6376)
                     /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(5734..5753)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6547..7135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7036..7055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
       61 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      121 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
      181 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
      241 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
      301 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
      361 cttttgcaaa aagcttgatt cttctgacac aacagtctcg aacttaagct gcagaagttg
      421 gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata
      481 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct
      541 tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca attacagctc
      601 ttaaggctag agtacttaat acgactcact ataggctagc caccatggct tccaaggtgt
      661 acgaccccga gcaacgcaaa cgcatgatca ctgggcctca gtggtgggct cgctgcaagc
      721 aaatgaacgt gctggactcc ttcatcaact actatgattc cgagaagcac gccgagaacg
      781 ccgtgatttt tctgcatggt aacgctgcct ccagctacct gtggaggcac gtcgtgcctc
      841 acatcgagcc cgtggctaga tgcatcatcc ctgatctgat cggaatgggt aagtccggca
      901 agagcgggaa tggctcatat cgcctcctgg atcactacaa gtacctcacc gcttggttcg
      961 agctgctgaa ccttccaaag aaaatcatct ttgtgggcca cgactggggg gcttgtctgg
     1021 cctttcacta ctcctacgag caccaagaca agatcaaggc catcgtccat gctgagagtg
     1081 tcgtggacgt gatcgagtcc tgggacgagt ggcctgacat cgaggaggat atcgccctga
     1141 tcaagagcga agagggcgag aaaatggtgc ttgagaataa cttcttcgtc gagaccatgc
     1201 tcccaagcaa gatcatgcgg aaactggagc ctgaggagtt cgctgcctac ctggagccat
     1261 tcaaggagaa gggcgaggtt agacggccta ccctctcctg gcctcgcgag atccctctcg
     1321 ttaagggagg caagcccgac gtcgtccaga ttgtccgcaa ctacaacgcc taccttcggg
     1381 ccagcgacga tctgcctaag atgttcatcg agtccgaccc tgggttcttt tccaacgcta
     1441 ttgtcgaggg agctaagaag ttccctaaca ccgagttcgt gaaggtgaag ggcctccact
     1501 tcagccagga ggacgctcca gatgaaatgg gtaagtacat caagagcttc gtggagcgcg
     1561 tgctgaagaa cgagcagtaa ttctaggcga tcgctcgagc ctggagcact gtggggacgg
     1621 actccgcctg ctgttcccgc gctgagctac aggactcccg agtgtgagcc gcggttcctc
     1681 tgttgcagcg cagccgtgtg tgctgtggag ccgaggccgt cctgcaggaa gccgcgtgac
     1741 tcccgactcc tccctgtgct ctctggctct ggactgtgac tgcgcctgga ttctgccatt
     1801 gcgacacatt tttgtgcctt tcagcccctg gtgtctgcag tgggggattt aaggcacccg
     1861 cttccacttc tttcttgttt ggagttttct gttggaaccg ccggcgttgg ctccgaagac
     1921 ttagcgacgc cactggcggc accttctcct gcgcccagtg atgtttccac ggtgcctgta
     1981 cacagccgag cagcatttcc gttgaaggac ttgcatcccc attgcgggca gtgctggacg
     2041 tgtcccggag acccaccggg agggcaccgc catgccttgt acccccaccg tgcaggttgt
     2101 ggccggtttt ctccgcaggt tgaacatgga aataaaagca aacttgtatg actcgagccc
     2161 ccccctaccc tcaacctcaa acccccacca aaaccacaca acacaaacca tcccacaaac
     2221 acccaaaact aacccccacc aaccccaaac accatcagag ctcactccac cccccccact
     2281 acacacctaa tttcttccca ctatctccaa ccactcccta accataaaac cctccccaca
     2341 acctaaccaa accaccaaaa ctagtttaaa cctagagcgg ccgctggccg caataaaata
     2401 tctttatttt cattacatct gtgtgttggt tttttgtgtg aggatctaaa tgagtcttcg
     2461 gacctcgcgg gggccgctta agcggtggtt agggtttgtc tgacgcgggg ggagggggaa
     2521 ggaacgaaac actctcattc ggaggcggct cggggtttgg tcttggtggc cacgggcacg
     2581 cagaagagcg ccgcgatcct cttaagcacc cccccgccct ccgtggaggc gggggtttgg
     2641 tcggcgggtg gtaactggcg ggccgctgac tcgggcgggt cgcgcgcccc agagtgtgac
     2701 cttttcggtc tgctcgcaga cccccgggcg gcgccgccgc ggcggcgacg ggctcgctgg
     2761 gtcctaggct ccatggggac cgtatacgtg gacaggctct ggagcatccg cacgactgcg
     2821 gtgatattac cggagacctt ctgcgggacg agccgggtca cgcggctgac gcggagcgtc
     2881 cgttgggcga caaacaccag gacggggcac aggtacacta tcttgtcacc cggaggcgcg
     2941 agggactgca ggagcttcag ggagtggcgc agctgcttca tccccgtggc ccgttgctcg
     3001 cgtttgctgg cggtgtcccc ggaagaaata tatttgcatg tctttagttc tatgatgaca
     3061 caaaccccgc ccagcgtctt gtcattggcg aattcgaaca cgcagatgca gtcggggcgg
     3121 cgcggtccca ggtccacttc gcatattaag gtgacgcgtg tggcctcgaa caccgagcga
     3181 ccctgcagcg acccgcttaa aagcttgatt cttctgacac aacagtctcg aacttaagct
     3241 gcagaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag
     3301 gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac
     3361 ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca
     3421 attacagctc ttaaggctag agtacttaat acgactcact ataggctagc aaagccacca
     3481 tggccgatgc taagaacatt aagaagggcc ctgctccctt ctaccctctg gaggatggca
     3541 ccgctggcga gcagctgcac aaggccatga agaggtatgc cctggtgcct ggcaccattg
     3601 ccttcaccga tgcccacatt gaggtggaca tcacctatgc cgagtacttc gagatgtctg
     3661 tgcgcctggc cgaggccatg aagaggtacg gcctgaacac caaccaccgc atcgtggtgt
     3721 gctctgagaa ctctctgcag ttcttcatgc cagtgctggg cgccctgttc atcggagtgg
     3781 ccgtggcccc tgctaacgac atttacaacg agcgcgagct gctgaacagc atgggcattt
     3841 ctcagcctac cgtggtgttc gtgtctaaga agggcctgca gaagatcctg aacgtgcaga
     3901 agaagctgcc tatcatccag aagatcatca tcatggactc taagaccgac taccagggct
     3961 tccagagcat gtacacattc gtgacatctc atctgcctcc tggcttcaac gagtacgact
     4021 tcgtgccaga gtctttcgac agggacaaaa ccattgccct gatcatgaac agctctgggt
     4081 ctaccggcct gcctaagggc gtggccctgc ctcatcgcac cgcctgtgtg cgcttctctc
     4141 acgcccgcga ccctattttc ggcaaccaga tcatccccga caccgctatt ctgagcgtgg
     4201 tgccattcca ccacggcttc ggcatgttca ccaccctggg ctacctgatt tgcggctttc
     4261 gggtggtgct gatgtaccgc ttcgaggagg agctgttcct gcgcagcctg caagactaca
     4321 aaattcagtc tgccctgctg gtgccaaccc tgttcagctt cttcgctaag agcaccctga
     4381 tcgacaagta cgacctgtct aacctgcacg agattgcctc tggcggcgcc ccactgtcta
     4441 aggaggtggg cgaagccgtg gccaagcgct ttcatctgcc aggcatccgc cagggctacg
     4501 gcctgaccga gacaaccagc gccattctga ttaccccaga gggcgacgac aagcctggcg
     4561 ccgtgggcaa ggtggtgcca ttcttcgagg ccaaggtggt ggacctggac accggcaaga
     4621 ccctgggagt gaaccagcgc ggcgagctgt gtgtgcgcgg ccctatgatt atgtccggct
     4681 acgtgaataa ccctgaggcc acaaacgccc tgatcgacaa ggacggctgg ctgcactctg
     4741 gcgacattgc ctactgggac gaggacgagc acttcttcat cgtggaccgc ctgaagtctc
     4801 tgatcaagta caagggctac caggtggccc cagccgagct ggagtctatc ctgctgcagc
     4861 accctaacat tttcgacgcc ggagtggccg gcctgcccga cgacgatgcc ggcgagctgc
     4921 ctgccgccgt cgtcgtgctg gaacacggca agaccatgac cgagaaggag atcgtggact
     4981 atgtggccag ccaggtgaca accgccaaga agctgcgcgg cggagtggtg ttcgtggacg
     5041 aggtgcccaa gggcctgacc ggcaagctgg acgcccgcaa gatccgcgag atcctgatca
     5101 aggctaagaa aggcggcaag atcgccgtgt aatgcggccg cttcgagcag acatgataag
     5161 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     5221 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     5281 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta
     5341 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccaggtg gcacttttcg
     5401 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5461 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5521 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5581 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5641 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5701 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5761 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5821 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5881 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5941 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6001 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6061 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6121 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6181 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6241 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6301 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6361 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6421 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6481 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6541 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6601 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6661 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6721 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6781 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6841 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6901 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6961 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7021 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7081 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7141 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatggctcg
     7201 acagatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt g
//
