LOCUS       sequence-269667-        6575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EIF3B (465-552) 3'UTR with the main hairpin restored 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 6575)
  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 6575)
  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-88nt-Restored-Hairpin
FEATURES             Location/Qualifiers
     source          1..6575
                     /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    1769..1817
                     /note="synthetic polyadenylation signal"
     promoter        1826..2577
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2647..2779
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        2824..2842
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2851..2860
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2857..4509
                     /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             3816..3824
                     /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    4534..4655
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4545..4564)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4599..4618
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4788..4892
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4893..4961,4962..5753)
                     /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(5111..5130)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5924..6512
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6413..6432
                     /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 aggctggcgg tacccttgta cccccaccgt
     1681 gcaggtacac cggccatttc tccgcaggtt gaacatggaa ataaaagcaa acttgtatga
     1741 gtttaaacct agagcggccg ctggccgcaa taaaatatct ttattttcat tacatctgtg
     1801 tgttggtttt ttgtgtgagg atctaaatga gtcttcggac ctcgcggggg ccgcttaagc
     1861 ggtggttagg gtttgtctga cgcgggggga gggggaagga acgaaacact ctcattcgga
     1921 ggcggctcgg ggtttggtct tggtggccac gggcacgcag aagagcgccg cgatcctctt
     1981 aagcaccccc ccgccctccg tggaggcggg ggtttggtcg gcgggtggta actggcgggc
     2041 cgctgactcg ggcgggtcgc gcgccccaga gtgtgacctt ttcggtctgc tcgcagaccc
     2101 ccgggcggcg ccgccgcggc ggcgacgggc tcgctgggtc ctaggctcca tggggaccgt
     2161 atacgtggac aggctctgga gcatccgcac gactgcggtg atattaccgg agaccttctg
     2221 cgggacgagc cgggtcacgc ggctgacgcg gagcgtccgt tgggcgacaa acaccaggac
     2281 ggggcacagg tacactatct tgtcacccgg aggcgcgagg gactgcagga gcttcaggga
     2341 gtggcgcagc tgcttcatcc ccgtggcccg ttgctcgcgt ttgctggcgg tgtccccgga
     2401 agaaatatat ttgcatgtct ttagttctat gatgacacaa accccgccca gcgtcttgtc
     2461 attggcgaat tcgaacacgc agatgcagtc ggggcggcgc ggtcccaggt ccacttcgca
     2521 tattaaggtg acgcgtgtgg cctcgaacac cgagcgaccc tgcagcgacc cgcttaaaag
     2581 cttgattctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc gtgaggcact
     2641 gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg
     2701 tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac
     2761 tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta aggctagagt
     2821 acttaatacg actcactata ggctagcaaa gccaccatgg ccgatgctaa gaacattaag
     2881 aagggccctg ctcccttcta ccctctggag gatggcaccg ctggcgagca gctgcacaag
     2941 gccatgaaga ggtatgccct ggtgcctggc accattgcct tcaccgatgc ccacattgag
     3001 gtggacatca cctatgccga gtacttcgag atgtctgtgc gcctggccga ggccatgaag
     3061 aggtacggcc tgaacaccaa ccaccgcatc gtggtgtgct ctgagaactc tctgcagttc
     3121 ttcatgccag tgctgggcgc cctgttcatc ggagtggccg tggcccctgc taacgacatt
     3181 tacaacgagc gcgagctgct gaacagcatg ggcatttctc agcctaccgt ggtgttcgtg
     3241 tctaagaagg gcctgcagaa gatcctgaac gtgcagaaga agctgcctat catccagaag
     3301 atcatcatca tggactctaa gaccgactac cagggcttcc agagcatgta cacattcgtg
     3361 acatctcatc tgcctcctgg cttcaacgag tacgacttcg tgccagagtc tttcgacagg
     3421 gacaaaacca ttgccctgat catgaacagc tctgggtcta ccggcctgcc taagggcgtg
     3481 gccctgcctc atcgcaccgc ctgtgtgcgc ttctctcacg cccgcgaccc tattttcggc
     3541 aaccagatca tccccgacac cgctattctg agcgtggtgc cattccacca cggcttcggc
     3601 atgttcacca ccctgggcta cctgatttgc ggctttcggg tggtgctgat gtaccgcttc
     3661 gaggaggagc tgttcctgcg cagcctgcaa gactacaaaa ttcagtctgc cctgctggtg
     3721 ccaaccctgt tcagcttctt cgctaagagc accctgatcg acaagtacga cctgtctaac
     3781 ctgcacgaga ttgcctctgg cggcgcccca ctgtctaagg aggtgggcga agccgtggcc
     3841 aagcgctttc atctgccagg catccgccag ggctacggcc tgaccgagac aaccagcgcc
     3901 attctgatta ccccagaggg cgacgacaag cctggcgccg tgggcaaggt ggtgccattc
     3961 ttcgaggcca aggtggtgga cctggacacc ggcaagaccc tgggagtgaa ccagcgcggc
     4021 gagctgtgtg tgcgcggccc tatgattatg tccggctacg tgaataaccc tgaggccaca
     4081 aacgccctga tcgacaagga cggctggctg cactctggcg acattgccta ctgggacgag
     4141 gacgagcact tcttcatcgt ggaccgcctg aagtctctga tcaagtacaa gggctaccag
     4201 gtggccccag ccgagctgga gtctatcctg ctgcagcacc ctaacatttt cgacgccgga
     4261 gtggccggcc tgcccgacga cgatgccggc gagctgcctg ccgccgtcgt cgtgctggaa
     4321 cacggcaaga ccatgaccga gaaggagatc gtggactatg tggccagcca ggtgacaacc
     4381 gccaagaagc tgcgcggcgg agtggtgttc gtggacgagg tgcccaaggg cctgaccggc
     4441 aagctggacg cccgcaagat ccgcgagatc ctgatcaagg ctaagaaagg cggcaagatc
     4501 gccgtgtaat gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     4561 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     4621 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     4681 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     4741 gtggtaaaat cgataaggat ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4801 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4861 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4921 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4981 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5041 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5101 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5161 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5221 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5281 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5341 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5401 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5461 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5521 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5581 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5641 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5701 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5761 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5821 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5881 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5941 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6001 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6061 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6121 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6181 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6241 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6301 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6361 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6421 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6481 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6541 tcctggcctt ttgctggcct tttgctcaca tggct
//
