LOCUS       sequence-269576-        7039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EIF3B 3'UTR (NM_001037283.1) 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 7039)
  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 7039)
  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
FEATURES             Location/Qualifiers
     source          1..7039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          79..211
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        256..274
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      278..287
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             284..1219
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1819..1867
                     /note="synthetic polyadenylation signal"
     promoter        1876..2627
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2697..2829
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        2874..2892
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2901..2910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2907..4559
                     /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             3866..3874
                     /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    4584..4705
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4595..4614)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4649..4668
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4838..4942
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4943..5011,5012..5803)
                     /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(5161..5180)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5974..6562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6463..6482
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(6691..7039,1..9)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6691..6882
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6714..6734)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6899..7034
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ttttgcaaaa agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg
       61 tcgtgaggca ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag
      121 aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt
      181 actgacatcc actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct
      241 taaggctaga gtacttaata cgactcacta taggctagcc accatggctt ccaaggtgta
      301 cgaccccgag caacgcaaac gcatgatcac tgggcctcag tggtgggctc gctgcaagca
      361 aatgaacgtg ctggactcct tcatcaacta ctatgattcc gagaagcacg ccgagaacgc
      421 cgtgattttt ctgcatggta acgctgcctc cagctacctg tggaggcacg tcgtgcctca
      481 catcgagccc gtggctagat gcatcatccc tgatctgatc ggaatgggta agtccggcaa
      541 gagcgggaat ggctcatatc gcctcctgga tcactacaag tacctcaccg cttggttcga
      601 gctgctgaac cttccaaaga aaatcatctt tgtgggccac gactgggggg cttgtctggc
      661 ctttcactac tcctacgagc accaagacaa gatcaaggcc atcgtccatg ctgagagtgt
      721 cgtggacgtg atcgagtcct gggacgagtg gcctgacatc gaggaggata tcgccctgat
      781 caagagcgaa gagggcgaga aaatggtgct tgagaataac ttcttcgtcg agaccatgct
      841 cccaagcaag atcatgcgga aactggagcc tgaggagttc gctgcctacc tggagccatt
      901 caaggagaag ggcgaggtta gacggcctac cctctcctgg cctcgcgaga tccctctcgt
      961 taagggaggc aagcccgacg tcgtccagat tgtccgcaac tacaacgcct accttcgggc
     1021 cagcgacgat ctgcctaaga tgttcatcga gtccgaccct gggttctttt ccaacgctat
     1081 tgtcgaggga gctaagaagt tccctaacac cgagttcgtg aaggtgaagg gcctccactt
     1141 cagccaggag gacgctccag atgaaatggg taagtacatc aagagcttcg tggagcgcgt
     1201 gctgaagaac gagcagtaat tctaggcgat cgctcgagcc tggagcactg tggggacgga
     1261 ctccgcctgc tgttcccgcg ctgagctaca ggactcccga gtgtgagccg cggttcctct
     1321 gttgcagcgc agccgtgtgt gctgtggagc cgaggccgtc ctgcaggaag ccgcgtgact
     1381 cccgactcct ccctgtgctc tctggctctg gactgtgact gcgcctggat tctgccattg
     1441 cgacacattt ttgtgccttt cagcccctgg tgtctgcagt gggggattta aggcacccgc
     1501 ttccacttct ttcttgtttg gagttttctg ttggaaccgc cggcgttggc tccgaagact
     1561 tagcgacgcc actggcggca ccttctcctg cgcccagtga tgtttccacg gtgcctgtac
     1621 acagccgagc agcatttccg ttgaaggact tgcatcccca ttgcgggcag tgctggacgt
     1681 gtcccggaga cccaccggga gggcaccgcc atgccttgta cccccaccgt gcaggttgtg
     1741 gccggttttc tccgcaggtt gaacatggaa ataaaagcaa acttgtatga gtttaaacct
     1801 agagcggccg ctggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt
     1861 ttgtgtgagg atctaaatga gtcttcggac ctcgcggggg ccgcttaagc ggtggttagg
     1921 gtttgtctga cgcgggggga gggggaagga acgaaacact ctcattcgga ggcggctcgg
     1981 ggtttggtct tggtggccac gggcacgcag aagagcgccg cgatcctctt aagcaccccc
     2041 ccgccctccg tggaggcggg ggtttggtcg gcgggtggta actggcgggc cgctgactcg
     2101 ggcgggtcgc gcgccccaga gtgtgacctt ttcggtctgc tcgcagaccc ccgggcggcg
     2161 ccgccgcggc ggcgacgggc tcgctgggtc ctaggctcca tggggaccgt atacgtggac
     2221 aggctctgga gcatccgcac gactgcggtg atattaccgg agaccttctg cgggacgagc
     2281 cgggtcacgc ggctgacgcg gagcgtccgt tgggcgacaa acaccaggac ggggcacagg
     2341 tacactatct tgtcacccgg aggcgcgagg gactgcagga gcttcaggga gtggcgcagc
     2401 tgcttcatcc ccgtggcccg ttgctcgcgt ttgctggcgg tgtccccgga agaaatatat
     2461 ttgcatgtct ttagttctat gatgacacaa accccgccca gcgtcttgtc attggcgaat
     2521 tcgaacacgc agatgcagtc ggggcggcgc ggtcccaggt ccacttcgca tattaaggtg
     2581 acgcgtgtgg cctcgaacac cgagcgaccc tgcagcgacc cgcttaaaag cttgattctt
     2641 ctgacacaac agtctcgaac ttaagctgca gaagttggtc gtgaggcact gggcaggtaa
     2701 gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg tcgagacaga
     2761 gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac tttgcctttc
     2821 tctccacagg tgtccactcc cagttcaatt acagctctta aggctagagt acttaatacg
     2881 actcactata ggctagcaaa gccaccatgg ccgatgctaa gaacattaag aagggccctg
     2941 ctcccttcta ccctctggag gatggcaccg ctggcgagca gctgcacaag gccatgaaga
     3001 ggtatgccct ggtgcctggc accattgcct tcaccgatgc ccacattgag gtggacatca
     3061 cctatgccga gtacttcgag atgtctgtgc gcctggccga ggccatgaag aggtacggcc
     3121 tgaacaccaa ccaccgcatc gtggtgtgct ctgagaactc tctgcagttc ttcatgccag
     3181 tgctgggcgc cctgttcatc ggagtggccg tggcccctgc taacgacatt tacaacgagc
     3241 gcgagctgct gaacagcatg ggcatttctc agcctaccgt ggtgttcgtg tctaagaagg
     3301 gcctgcagaa gatcctgaac gtgcagaaga agctgcctat catccagaag atcatcatca
     3361 tggactctaa gaccgactac cagggcttcc agagcatgta cacattcgtg acatctcatc
     3421 tgcctcctgg cttcaacgag tacgacttcg tgccagagtc tttcgacagg gacaaaacca
     3481 ttgccctgat catgaacagc tctgggtcta ccggcctgcc taagggcgtg gccctgcctc
     3541 atcgcaccgc ctgtgtgcgc ttctctcacg cccgcgaccc tattttcggc aaccagatca
     3601 tccccgacac cgctattctg agcgtggtgc cattccacca cggcttcggc atgttcacca
     3661 ccctgggcta cctgatttgc ggctttcggg tggtgctgat gtaccgcttc gaggaggagc
     3721 tgttcctgcg cagcctgcaa gactacaaaa ttcagtctgc cctgctggtg ccaaccctgt
     3781 tcagcttctt cgctaagagc accctgatcg acaagtacga cctgtctaac ctgcacgaga
     3841 ttgcctctgg cggcgcccca ctgtctaagg aggtgggcga agccgtggcc aagcgctttc
     3901 atctgccagg catccgccag ggctacggcc tgaccgagac aaccagcgcc attctgatta
     3961 ccccagaggg cgacgacaag cctggcgccg tgggcaaggt ggtgccattc ttcgaggcca
     4021 aggtggtgga cctggacacc ggcaagaccc tgggagtgaa ccagcgcggc gagctgtgtg
     4081 tgcgcggccc tatgattatg tccggctacg tgaataaccc tgaggccaca aacgccctga
     4141 tcgacaagga cggctggctg cactctggcg acattgccta ctgggacgag gacgagcact
     4201 tcttcatcgt ggaccgcctg aagtctctga tcaagtacaa gggctaccag gtggccccag
     4261 ccgagctgga gtctatcctg ctgcagcacc ctaacatttt cgacgccgga gtggccggcc
     4321 tgcccgacga cgatgccggc gagctgcctg ccgccgtcgt cgtgctggaa cacggcaaga
     4381 ccatgaccga gaaggagatc gtggactatg tggccagcca ggtgacaacc gccaagaagc
     4441 tgcgcggcgg agtggtgttc gtggacgagg tgcccaaggg cctgaccggc aagctggacg
     4501 cccgcaagat ccgcgagatc ctgatcaagg ctaagaaagg cggcaagatc gccgtgtaat
     4561 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     4621 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     4681 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     4741 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat
     4801 cgataaggat ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4861 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4921 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4981 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5041 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5101 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5161 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5221 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5281 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5341 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5401 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5461 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5521 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5581 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5641 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5701 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5761 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5821 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5881 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5941 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6001 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6061 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6121 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6181 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6241 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6301 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6361 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6421 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6481 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6541 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6601 ttgctggcct tttgctcaca tggctcgaca gatctgcgca gcaccatggc ctgaaataac
     6661 ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt
     6721 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     6781 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     6841 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     6901 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6961 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     7021 cttttttgga ggcctaggc
//
