LOCUS       sequence-269951-        6787 bp    DNA     circular SYN 15-APR-2025
DEFINITION  EIF3B (465-552) 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 6787)
  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 6787)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-465-552-Unstructured
FEATURES             Location/Qualifiers
     source          1..6787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        91..448
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        91..282
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(114..134)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      299..434
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          518..650
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        695..713
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      717..726
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             723..1658
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2006..2054
                     /note="synthetic polyadenylation signal"
     promoter        2063..2814
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2884..3016
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        3061..3079
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3088..3097
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3094..4746
                     /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             4053..4061
                     /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    4771..4892
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4782..4801)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4836..4855
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5025..5129
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5130..5198,5199..5990)
                     /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(5348..5367)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6161..6749
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6650..6669
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttgctggcct tttgctcaca tggctcgaca gatctgcgca gcaccatggc ctgaaataac
       61 ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt
      121 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
      181 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
      241 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
      301 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
      361 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
      421 cttttttgga ggcctaggct tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa
      481 cttaagctgc agaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca
      541 ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg
      601 ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag gtgtccactc
      661 ccagttcaat tacagctctt aaggctagag tacttaatac gactcactat aggctagcca
      721 ccatggcttc caaggtgtac gaccccgagc aacgcaaacg catgatcact gggcctcagt
      781 ggtgggctcg ctgcaagcaa atgaacgtgc tggactcctt catcaactac tatgattccg
      841 agaagcacgc cgagaacgcc gtgatttttc tgcatggtaa cgctgcctcc agctacctgt
      901 ggaggcacgt cgtgcctcac atcgagcccg tggctagatg catcatccct gatctgatcg
      961 gaatgggtaa gtccggcaag agcgggaatg gctcatatcg cctcctggat cactacaagt
     1021 acctcaccgc ttggttcgag ctgctgaacc ttccaaagaa aatcatcttt gtgggccacg
     1081 actggggggc ttgtctggcc tttcactact cctacgagca ccaagacaag atcaaggcca
     1141 tcgtccatgc tgagagtgtc gtggacgtga tcgagtcctg ggacgagtgg cctgacatcg
     1201 aggaggatat cgccctgatc aagagcgaag agggcgagaa aatggtgctt gagaataact
     1261 tcttcgtcga gaccatgctc ccaagcaaga tcatgcggaa actggagcct gaggagttcg
     1321 ctgcctacct ggagccattc aaggagaagg gcgaggttag acggcctacc ctctcctggc
     1381 ctcgcgagat ccctctcgtt aagggaggca agcccgacgt cgtccagatt gtccgcaact
     1441 acaacgccta ccttcgggcc agcgacgatc tgcctaagat gttcatcgag tccgaccctg
     1501 ggttcttttc caacgctatt gtcgagggag ctaagaagtt ccctaacacc gagttcgtga
     1561 aggtgaaggg cctccacttc agccaggagg acgctccaga tgaaatgggt aagtacatca
     1621 agagcttcgt ggagcgcgtg ctgaagaacg agcagtaatt ctaggcgatc gctcgaggca
     1681 ccgccatgcc ttgtaccccc accgtgcagg ttgtggccgg ttttctccgc aggttgaaca
     1741 tggaaataaa agcaaacttg tatgactcga gcccccccct accctcaacc tcaaaccccc
     1801 accaaaacca cacaacacaa accatcccac aaacacccaa aactaacccc caccaacccc
     1861 aaacaccatc agagctcact ccaccccccc cactacacac ctaatttctt cccactatct
     1921 ccaaccactc cctaaccata aaaccctccc cacaacctaa ccaaaccacc aaaactagtt
     1981 taaacctaga gcggccgctg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     2041 tggttttttg tgtgaggatc taaatgagtc ttcggacctc gcgggggccg cttaagcggt
     2101 ggttagggtt tgtctgacgc ggggggaggg ggaaggaacg aaacactctc attcggaggc
     2161 ggctcggggt ttggtcttgg tggccacggg cacgcagaag agcgccgcga tcctcttaag
     2221 cacccccccg ccctccgtgg aggcgggggt ttggtcggcg ggtggtaact ggcgggccgc
     2281 tgactcgggc gggtcgcgcg ccccagagtg tgaccttttc ggtctgctcg cagacccccg
     2341 ggcggcgccg ccgcggcggc gacgggctcg ctgggtccta ggctccatgg ggaccgtata
     2401 cgtggacagg ctctggagca tccgcacgac tgcggtgata ttaccggaga ccttctgcgg
     2461 gacgagccgg gtcacgcggc tgacgcggag cgtccgttgg gcgacaaaca ccaggacggg
     2521 gcacaggtac actatcttgt cacccggagg cgcgagggac tgcaggagct tcagggagtg
     2581 gcgcagctgc ttcatccccg tggcccgttg ctcgcgtttg ctggcggtgt ccccggaaga
     2641 aatatatttg catgtcttta gttctatgat gacacaaacc ccgcccagcg tcttgtcatt
     2701 ggcgaattcg aacacgcaga tgcagtcggg gcggcgcggt cccaggtcca cttcgcatat
     2761 taaggtgacg cgtgtggcct cgaacaccga gcgaccctgc agcgacccgc ttaaaagctt
     2821 gattcttctg acacaacagt ctcgaactta agctgcagaa gttggtcgtg aggcactggg
     2881 caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact gggcttgtcg
     2941 agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga catccacttt
     3001 gcctttctct ccacaggtgt ccactcccag ttcaattaca gctcttaagg ctagagtact
     3061 taatacgact cactataggc tagcaaagcc accatggccg atgctaagaa cattaagaag
     3121 ggccctgctc ccttctaccc tctggaggat ggcaccgctg gcgagcagct gcacaaggcc
     3181 atgaagaggt atgccctggt gcctggcacc attgccttca ccgatgccca cattgaggtg
     3241 gacatcacct atgccgagta cttcgagatg tctgtgcgcc tggccgaggc catgaagagg
     3301 tacggcctga acaccaacca ccgcatcgtg gtgtgctctg agaactctct gcagttcttc
     3361 atgccagtgc tgggcgccct gttcatcgga gtggccgtgg cccctgctaa cgacatttac
     3421 aacgagcgcg agctgctgaa cagcatgggc atttctcagc ctaccgtggt gttcgtgtct
     3481 aagaagggcc tgcagaagat cctgaacgtg cagaagaagc tgcctatcat ccagaagatc
     3541 atcatcatgg actctaagac cgactaccag ggcttccaga gcatgtacac attcgtgaca
     3601 tctcatctgc ctcctggctt caacgagtac gacttcgtgc cagagtcttt cgacagggac
     3661 aaaaccattg ccctgatcat gaacagctct gggtctaccg gcctgcctaa gggcgtggcc
     3721 ctgcctcatc gcaccgcctg tgtgcgcttc tctcacgccc gcgaccctat tttcggcaac
     3781 cagatcatcc ccgacaccgc tattctgagc gtggtgccat tccaccacgg cttcggcatg
     3841 ttcaccaccc tgggctacct gatttgcggc tttcgggtgg tgctgatgta ccgcttcgag
     3901 gaggagctgt tcctgcgcag cctgcaagac tacaaaattc agtctgccct gctggtgcca
     3961 accctgttca gcttcttcgc taagagcacc ctgatcgaca agtacgacct gtctaacctg
     4021 cacgagattg cctctggcgg cgccccactg tctaaggagg tgggcgaagc cgtggccaag
     4081 cgctttcatc tgccaggcat ccgccagggc tacggcctga ccgagacaac cagcgccatt
     4141 ctgattaccc cagagggcga cgacaagcct ggcgccgtgg gcaaggtggt gccattcttc
     4201 gaggccaagg tggtggacct ggacaccggc aagaccctgg gagtgaacca gcgcggcgag
     4261 ctgtgtgtgc gcggccctat gattatgtcc ggctacgtga ataaccctga ggccacaaac
     4321 gccctgatcg acaaggacgg ctggctgcac tctggcgaca ttgcctactg ggacgaggac
     4381 gagcacttct tcatcgtgga ccgcctgaag tctctgatca agtacaaggg ctaccaggtg
     4441 gccccagccg agctggagtc tatcctgctg cagcacccta acattttcga cgccggagtg
     4501 gccggcctgc ccgacgacga tgccggcgag ctgcctgccg ccgtcgtcgt gctggaacac
     4561 ggcaagacca tgaccgagaa ggagatcgtg gactatgtgg ccagccaggt gacaaccgcc
     4621 aagaagctgc gcggcggagt ggtgttcgtg gacgaggtgc ccaagggcct gaccggcaag
     4681 ctggacgccc gcaagatccg cgagatcctg atcaaggcta agaaaggcgg caagatcgcc
     4741 gtgtaatgcg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     4801 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     4861 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     4921 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     4981 gtaaaatcga taaggatcca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5041 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5101 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5161 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5221 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5281 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5341 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5401 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5461 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5521 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5581 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5641 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5701 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5761 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5821 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5881 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5941 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6001 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6061 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6121 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6181 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6241 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6301 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6361 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6421 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6481 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6541 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6601 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6661 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6721 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6781 tggcctt
//
