LOCUS       sequence-269959-        6964 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The Reverse Complement of EIF3B 3'UTR 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 6964)
  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 6964)
  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-Reverse-Complement
FEATURES             Location/Qualifiers
     source          1..6964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        66..423
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        66..257
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(89..109)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      274..409
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          493..625
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        670..688
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      692..701
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             698..1633
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2158..2206
                     /note="synthetic polyadenylation signal"
     promoter        2215..2966
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          3036..3168
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        3213..3231
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3240..3249
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3246..4898
                     /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             4205..4213
                     /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    4923..5044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4934..4953)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4988..5007
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5177..5281
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5282..5350,5351..6142)
                     /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(5500..5519)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6313..6901
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6802..6821
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgacagatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt
       61 accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
      121 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
      181 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
      241 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
      301 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
      361 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
      421 aaaaagcttg attcttctga cacaacagtc tcgaacttaa gctgcagaag ttggtcgtga
      481 ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg
      541 ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac
      601 atccactttg cctttctctc cacaggtgtc cactcccagt tcaattacag ctcttaaggc
      661 tagagtactt aatacgactc actataggct agccaccatg gcttccaagg tgtacgaccc
      721 cgagcaacgc aaacgcatga tcactgggcc tcagtggtgg gctcgctgca agcaaatgaa
      781 cgtgctggac tccttcatca actactatga ttccgagaag cacgccgaga acgccgtgat
      841 ttttctgcat ggtaacgctg cctccagcta cctgtggagg cacgtcgtgc ctcacatcga
      901 gcccgtggct agatgcatca tccctgatct gatcggaatg ggtaagtccg gcaagagcgg
      961 gaatggctca tatcgcctcc tggatcacta caagtacctc accgcttggt tcgagctgct
     1021 gaaccttcca aagaaaatca tctttgtggg ccacgactgg ggggcttgtc tggcctttca
     1081 ctactcctac gagcaccaag acaagatcaa ggccatcgtc catgctgaga gtgtcgtgga
     1141 cgtgatcgag tcctgggacg agtggcctga catcgaggag gatatcgccc tgatcaagag
     1201 cgaagagggc gagaaaatgg tgcttgagaa taacttcttc gtcgagacca tgctcccaag
     1261 caagatcatg cggaaactgg agcctgagga gttcgctgcc tacctggagc cattcaagga
     1321 gaagggcgag gttagacggc ctaccctctc ctggcctcgc gagatccctc tcgttaaggg
     1381 aggcaagccc gacgtcgtcc agattgtccg caactacaac gcctaccttc gggccagcga
     1441 cgatctgcct aagatgttca tcgagtccga ccctgggttc ttttccaacg ctattgtcga
     1501 gggagctaag aagttcccta acaccgagtt cgtgaaggtg aagggcctcc acttcagcca
     1561 ggaggacgct ccagatgaaa tgggtaagta catcaagagc ttcgtggagc gcgtgctgaa
     1621 gaacgagcag taattctagg cgatcgctcg agtcatacaa gtttgctttt atttccatgt
     1681 tcaacctgcg gagaaaaccg gccacaacct gcacggtggg ggtacaaggc atggcggcgc
     1741 cctcccggtg ggtctccggg acacgtccag cactgcccgc aatggggatg caagtccttc
     1801 aacggaaatg ctgctcggct gtgtacaggc accgtggaaa catcactggg cgcaggagaa
     1861 ggtgccgcca gtggcgtcgc taagtcttcg gagccaacgc cggcggttcc aacagaaaac
     1921 tccaaacaag aaagaagtgg aagcgggtgc cttaaatccc ccactgcaga caccaggggc
     1981 tgaaaggcac aaaaatgtgt cgcaatggca gaatccaggc gcagtcacag tccagagcca
     2041 gagagcacag ggaggaggcg ggagtcacgc ggcttcctgc aggacggcct cggctccaca
     2101 gcacacacgg ctgcgcacag tgctccaggg tttaaaccta gagcggccgc tggccgcaat
     2161 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgagga tctaaatgag
     2221 tcttcggacc tcgcgggggc cgcttaagcg gtggttaggg tttgtctgac gcggggggag
     2281 ggggaaggaa cgaaacactc tcattcggag gcggctcggg gtttggtctt ggtggccacg
     2341 ggcacgcaga agagcgccgc gatcctctta agcacccccc cgccctccgt ggaggcgggg
     2401 gtttggtcgg cgggtggtaa ctggcgggcc gctgactcgg gcgggtcgcg cgccccagag
     2461 tgtgaccttt tcggtctgct cgcagacccc cgggcggcgc cgccgcggcg gcgacgggct
     2521 cgctgggtcc taggctccat ggggaccgta tacgtggaca ggctctggag catccgcacg
     2581 actgcggtga tattaccgga gaccttctgc gggacgagcc gggtcacgcg gctgacgcgg
     2641 agcgtccgtt gggcgacaaa caccaggacg gggcacaggt acactatctt gtcacccgga
     2701 ggcgcgaggg actgcaggag cttcagggag tggcgcagct gcttcatccc cgtggcccgt
     2761 tgctcgcgtt tgctggcggt gtccccggaa gaaatatatt tgcatgtctt tagttctatg
     2821 atgacacaaa ccccgcccag cgtcttgtca ttggcgaatt cgaacacgca gatgcagtcg
     2881 gggcggcgcg gtcccaggtc cacttcgcat attaaggtga cgcgtgtggc ctcgaacacc
     2941 gagcgaccct gcagcgaccc gcttaaaagc ttgattcttc tgacacaaca gtctcgaact
     3001 taagctgcag aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt acaagacagg
     3061 tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat
     3121 aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt gtccactccc
     3181 agttcaatta cagctcttaa ggctagagta cttaatacga ctcactatag gctagcaaag
     3241 ccaccatggc cgatgctaag aacattaaga agggccctgc tcccttctac cctctggagg
     3301 atggcaccgc tggcgagcag ctgcacaagg ccatgaagag gtatgccctg gtgcctggca
     3361 ccattgcctt caccgatgcc cacattgagg tggacatcac ctatgccgag tacttcgaga
     3421 tgtctgtgcg cctggccgag gccatgaaga ggtacggcct gaacaccaac caccgcatcg
     3481 tggtgtgctc tgagaactct ctgcagttct tcatgccagt gctgggcgcc ctgttcatcg
     3541 gagtggccgt ggcccctgct aacgacattt acaacgagcg cgagctgctg aacagcatgg
     3601 gcatttctca gcctaccgtg gtgttcgtgt ctaagaaggg cctgcagaag atcctgaacg
     3661 tgcagaagaa gctgcctatc atccagaaga tcatcatcat ggactctaag accgactacc
     3721 agggcttcca gagcatgtac acattcgtga catctcatct gcctcctggc ttcaacgagt
     3781 acgacttcgt gccagagtct ttcgacaggg acaaaaccat tgccctgatc atgaacagct
     3841 ctgggtctac cggcctgcct aagggcgtgg ccctgcctca tcgcaccgcc tgtgtgcgct
     3901 tctctcacgc ccgcgaccct attttcggca accagatcat ccccgacacc gctattctga
     3961 gcgtggtgcc attccaccac ggcttcggca tgttcaccac cctgggctac ctgatttgcg
     4021 gctttcgggt ggtgctgatg taccgcttcg aggaggagct gttcctgcgc agcctgcaag
     4081 actacaaaat tcagtctgcc ctgctggtgc caaccctgtt cagcttcttc gctaagagca
     4141 ccctgatcga caagtacgac ctgtctaacc tgcacgagat tgcctctggc ggcgccccac
     4201 tgtctaagga ggtgggcgaa gccgtggcca agcgctttca tctgccaggc atccgccagg
     4261 gctacggcct gaccgagaca accagcgcca ttctgattac cccagagggc gacgacaagc
     4321 ctggcgccgt gggcaaggtg gtgccattct tcgaggccaa ggtggtggac ctggacaccg
     4381 gcaagaccct gggagtgaac cagcgcggcg agctgtgtgt gcgcggccct atgattatgt
     4441 ccggctacgt gaataaccct gaggccacaa acgccctgat cgacaaggac ggctggctgc
     4501 actctggcga cattgcctac tgggacgagg acgagcactt cttcatcgtg gaccgcctga
     4561 agtctctgat caagtacaag ggctaccagg tggccccagc cgagctggag tctatcctgc
     4621 tgcagcaccc taacattttc gacgccggag tggccggcct gcccgacgac gatgccggcg
     4681 agctgcctgc cgccgtcgtc gtgctggaac acggcaagac catgaccgag aaggagatcg
     4741 tggactatgt ggccagccag gtgacaaccg ccaagaagct gcgcggcgga gtggtgttcg
     4801 tggacgaggt gcccaagggc ctgaccggca agctggacgc ccgcaagatc cgcgagatcc
     4861 tgatcaaggc taagaaaggc ggcaagatcg ccgtgtaatg cggccgcttc gagcagacat
     4921 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     4981 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     5041 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     5101 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc caggtggcac
     5161 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5221 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5281 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5341 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5401 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5461 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5521 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5581 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5641 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5701 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5761 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5821 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5881 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5941 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6001 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6061 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6121 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6181 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6241 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6301 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6361 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6421 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6481 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6541 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6601 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6661 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6721 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6781 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6841 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6901 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6961 ggct
//
