LOCUS       sequence-263061-        6498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  psi-CHECK2 plasmid modified so both Renilla and Firefly luciferase 
            contain introns.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6498)
  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 6498)
  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: psi-CHECK3
FEATURES             Location/Qualifiers
     source          1..6498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        62..253
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(85..105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      270..405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          489..621
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             694..1629
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1688..1736
                     /note="synthetic polyadenylation signal"
     promoter        1745..2496
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2566..2698
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        2743..2761
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2770..2779
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2776..4428
                     /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             3735..3743
                     /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    4453..4574
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4464..4483)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4518..4537
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4707..4811
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4812..4880,4881..5672)
                     /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(5030..5049)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5843..6431
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6332..6351
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
       61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
      481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
      661 gtacttaata cgactcacta taggctagcc accatggctt ccaaggtgta cgaccccgag
      721 caacgcaaac gcatgatcac tgggcctcag tggtgggctc gctgcaagca aatgaacgtg
      781 ctggactcct tcatcaacta ctatgattcc gagaagcacg ccgagaacgc cgtgattttt
      841 ctgcatggta acgctgcctc cagctacctg tggaggcacg tcgtgcctca catcgagccc
      901 gtggctagat gcatcatccc tgatctgatc ggaatgggta agtccggcaa gagcgggaat
      961 ggctcatatc gcctcctgga tcactacaag tacctcaccg cttggttcga gctgctgaac
     1021 cttccaaaga aaatcatctt tgtgggccac gactgggggg cttgtctggc ctttcactac
     1081 tcctacgagc accaagacaa gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg
     1141 atcgagtcct gggacgagtg gcctgacatc gaggaggata tcgccctgat caagagcgaa
     1201 gagggcgaga aaatggtgct tgagaataac ttcttcgtcg agaccatgct cccaagcaag
     1261 atcatgcgga aactggagcc tgaggagttc gctgcctacc tggagccatt caaggagaag
     1321 ggcgaggtta gacggcctac cctctcctgg cctcgcgaga tccctctcgt taagggaggc
     1381 aagcccgacg tcgtccagat tgtccgcaac tacaacgcct accttcgggc cagcgacgat
     1441 ctgcctaaga tgttcatcga gtccgaccct gggttctttt ccaacgctat tgtcgaggga
     1501 gctaagaagt tccctaacac cgagttcgtg aaggtgaagg gcctccactt cagccaggag
     1561 gacgctccag atgaaatggg taagtacatc aagagcttcg tggagcgcgt gctgaagaac
     1621 gagcagtaat tctaggcgat cgctcgagcc cgggaattcg tttaaaccta gagcggccgc
     1681 tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgagga
     1741 tctaaatgag tcttcggacc tcgcgggggc cgcttaagcg gtggttaggg tttgtctgac
     1801 gcggggggag ggggaaggaa cgaaacactc tcattcggag gcggctcggg gtttggtctt
     1861 ggtggccacg ggcacgcaga agagcgccgc gatcctctta agcacccccc cgccctccgt
     1921 ggaggcgggg gtttggtcgg cgggtggtaa ctggcgggcc gctgactcgg gcgggtcgcg
     1981 cgccccagag tgtgaccttt tcggtctgct cgcagacccc cgggcggcgc cgccgcggcg
     2041 gcgacgggct cgctgggtcc taggctccat ggggaccgta tacgtggaca ggctctggag
     2101 catccgcacg actgcggtga tattaccgga gaccttctgc gggacgagcc gggtcacgcg
     2161 gctgacgcgg agcgtccgtt gggcgacaaa caccaggacg gggcacaggt acactatctt
     2221 gtcacccgga ggcgcgaggg actgcaggag cttcagggag tggcgcagct gcttcatccc
     2281 cgtggcccgt tgctcgcgtt tgctggcggt gtccccggaa gaaatatatt tgcatgtctt
     2341 tagttctatg atgacacaaa ccccgcccag cgtcttgtca ttggcgaatt cgaacacgca
     2401 gatgcagtcg gggcggcgcg gtcccaggtc cacttcgcat attaaggtga cgcgtgtggc
     2461 ctcgaacacc gagcgaccct gcagcgaccc gcttaaaagc ttgattcttc tgacacaaca
     2521 gtctcgaact taagctgcag aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt
     2581 acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg
     2641 cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt
     2701 gtccactccc agttcaatta cagctcttaa ggctagagta cttaatacga ctcactatag
     2761 gctagcaaag ccaccatggc cgatgctaag aacattaaga agggccctgc tcccttctac
     2821 cctctggagg atggcaccgc tggcgagcag ctgcacaagg ccatgaagag gtatgccctg
     2881 gtgcctggca ccattgcctt caccgatgcc cacattgagg tggacatcac ctatgccgag
     2941 tacttcgaga tgtctgtgcg cctggccgag gccatgaaga ggtacggcct gaacaccaac
     3001 caccgcatcg tggtgtgctc tgagaactct ctgcagttct tcatgccagt gctgggcgcc
     3061 ctgttcatcg gagtggccgt ggcccctgct aacgacattt acaacgagcg cgagctgctg
     3121 aacagcatgg gcatttctca gcctaccgtg gtgttcgtgt ctaagaaggg cctgcagaag
     3181 atcctgaacg tgcagaagaa gctgcctatc atccagaaga tcatcatcat ggactctaag
     3241 accgactacc agggcttcca gagcatgtac acattcgtga catctcatct gcctcctggc
     3301 ttcaacgagt acgacttcgt gccagagtct ttcgacaggg acaaaaccat tgccctgatc
     3361 atgaacagct ctgggtctac cggcctgcct aagggcgtgg ccctgcctca tcgcaccgcc
     3421 tgtgtgcgct tctctcacgc ccgcgaccct attttcggca accagatcat ccccgacacc
     3481 gctattctga gcgtggtgcc attccaccac ggcttcggca tgttcaccac cctgggctac
     3541 ctgatttgcg gctttcgggt ggtgctgatg taccgcttcg aggaggagct gttcctgcgc
     3601 agcctgcaag actacaaaat tcagtctgcc ctgctggtgc caaccctgtt cagcttcttc
     3661 gctaagagca ccctgatcga caagtacgac ctgtctaacc tgcacgagat tgcctctggc
     3721 ggcgccccac tgtctaagga ggtgggcgaa gccgtggcca agcgctttca tctgccaggc
     3781 atccgccagg gctacggcct gaccgagaca accagcgcca ttctgattac cccagagggc
     3841 gacgacaagc ctggcgccgt gggcaaggtg gtgccattct tcgaggccaa ggtggtggac
     3901 ctggacaccg gcaagaccct gggagtgaac cagcgcggcg agctgtgtgt gcgcggccct
     3961 atgattatgt ccggctacgt gaataaccct gaggccacaa acgccctgat cgacaaggac
     4021 ggctggctgc actctggcga cattgcctac tgggacgagg acgagcactt cttcatcgtg
     4081 gaccgcctga agtctctgat caagtacaag ggctaccagg tggccccagc cgagctggag
     4141 tctatcctgc tgcagcaccc taacattttc gacgccggag tggccggcct gcccgacgac
     4201 gatgccggcg agctgcctgc cgccgtcgtc gtgctggaac acggcaagac catgaccgag
     4261 aaggagatcg tggactatgt ggccagccag gtgacaaccg ccaagaagct gcgcggcgga
     4321 gtggtgttcg tggacgaggt gcccaagggc ctgaccggca agctggacgc ccgcaagatc
     4381 cgcgagatcc tgatcaaggc taagaaaggc ggcaagatcg ccgtgtaatg cggccgcttc
     4441 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     4501 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     4561 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     4621 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc
     4681 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4741 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4801 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4861 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4921 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4981 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5041 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5101 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5161 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5221 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5281 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5341 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5401 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5461 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5521 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5581 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5641 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5701 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5761 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5821 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5881 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5941 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6001 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6061 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6121 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6181 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6241 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6301 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6361 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6421 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6481 ttgctcacat ggctcgac
//
