LOCUS       sequence-269425-        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        61..418
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        61..252
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(84..104)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      269..404
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          488..620
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        665..683
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      687..696
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             693..1628
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1687..1735
                     /note="synthetic polyadenylation signal"
     promoter        1744..2495
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2565..2697
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        2742..2760
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2769..2778
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2775..4427
                     /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             3734..3742
                     /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    4452..4573
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4463..4482)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4517..4536
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4706..4810
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4811..4879,4880..5671)
                     /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(5029..5048)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5842..6430
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6331..6350
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt
       61 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
      121 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
      181 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      241 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
      301 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
      361 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
      421 gcttgattct tctgacacaa cagtctcgaa cttaagctgc agaagttggt cgtgaggcac
      481 tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt
      541 gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca
      601 ctttgccttt ctctccacag gtgtccactc ccagttcaat tacagctctt aaggctagag
      661 tacttaatac gactcactat aggctagcca ccatggcttc caaggtgtac gaccccgagc
      721 aacgcaaacg catgatcact gggcctcagt ggtgggctcg ctgcaagcaa atgaacgtgc
      781 tggactcctt catcaactac tatgattccg agaagcacgc cgagaacgcc gtgatttttc
      841 tgcatggtaa cgctgcctcc agctacctgt ggaggcacgt cgtgcctcac atcgagcccg
      901 tggctagatg catcatccct gatctgatcg gaatgggtaa gtccggcaag agcgggaatg
      961 gctcatatcg cctcctggat cactacaagt acctcaccgc ttggttcgag ctgctgaacc
     1021 ttccaaagaa aatcatcttt gtgggccacg actggggggc ttgtctggcc tttcactact
     1081 cctacgagca ccaagacaag atcaaggcca tcgtccatgc tgagagtgtc gtggacgtga
     1141 tcgagtcctg ggacgagtgg cctgacatcg aggaggatat cgccctgatc aagagcgaag
     1201 agggcgagaa aatggtgctt gagaataact tcttcgtcga gaccatgctc ccaagcaaga
     1261 tcatgcggaa actggagcct gaggagttcg ctgcctacct ggagccattc aaggagaagg
     1321 gcgaggttag acggcctacc ctctcctggc ctcgcgagat ccctctcgtt aagggaggca
     1381 agcccgacgt cgtccagatt gtccgcaact acaacgccta ccttcgggcc agcgacgatc
     1441 tgcctaagat gttcatcgag tccgaccctg ggttcttttc caacgctatt gtcgagggag
     1501 ctaagaagtt ccctaacacc gagttcgtga aggtgaaggg cctccacttc agccaggagg
     1561 acgctccaga tgaaatgggt aagtacatca agagcttcgt ggagcgcgtg ctgaagaacg
     1621 agcagtaatt ctaggcgatc gctcgagccc gggaattcgt ttaaacctag agcggccgct
     1681 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaggat
     1741 ctaaatgagt cttcggacct cgcgggggcc gcttaagcgg tggttagggt ttgtctgacg
     1801 cggggggagg gggaaggaac gaaacactct cattcggagg cggctcgggg tttggtcttg
     1861 gtggccacgg gcacgcagaa gagcgccgcg atcctcttaa gcaccccccc gccctccgtg
     1921 gaggcggggg tttggtcggc gggtggtaac tggcgggccg ctgactcggg cgggtcgcgc
     1981 gccccagagt gtgacctttt cggtctgctc gcagaccccc gggcggcgcc gccgcggcgg
     2041 cgacgggctc gctgggtcct aggctccatg gggaccgtat acgtggacag gctctggagc
     2101 atccgcacga ctgcggtgat attaccggag accttctgcg ggacgagccg ggtcacgcgg
     2161 ctgacgcgga gcgtccgttg ggcgacaaac accaggacgg ggcacaggta cactatcttg
     2221 tcacccggag gcgcgaggga ctgcaggagc ttcagggagt ggcgcagctg cttcatcccc
     2281 gtggcccgtt gctcgcgttt gctggcggtg tccccggaag aaatatattt gcatgtcttt
     2341 agttctatga tgacacaaac cccgcccagc gtcttgtcat tggcgaattc gaacacgcag
     2401 atgcagtcgg ggcggcgcgg tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc
     2461 tcgaacaccg agcgaccctg cagcgacccg cttaaaagct tgattcttct gacacaacag
     2521 tctcgaactt aagctgcaga agttggtcgt gaggcactgg gcaggtaagt atcaaggtta
     2581 caagacaggt ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc
     2641 gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg
     2701 tccactccca gttcaattac agctcttaag gctagagtac ttaatacgac tcactatagg
     2761 ctagcaaagc caccatggcc gatgctaaga acattaagaa gggccctgct cccttctacc
     2821 ctctggagga tggcaccgct ggcgagcagc tgcacaaggc catgaagagg tatgccctgg
     2881 tgcctggcac cattgccttc accgatgccc acattgaggt ggacatcacc tatgccgagt
     2941 acttcgagat gtctgtgcgc ctggccgagg ccatgaagag gtacggcctg aacaccaacc
     3001 accgcatcgt ggtgtgctct gagaactctc tgcagttctt catgccagtg ctgggcgccc
     3061 tgttcatcgg agtggccgtg gcccctgcta acgacattta caacgagcgc gagctgctga
     3121 acagcatggg catttctcag cctaccgtgg tgttcgtgtc taagaagggc ctgcagaaga
     3181 tcctgaacgt gcagaagaag ctgcctatca tccagaagat catcatcatg gactctaaga
     3241 ccgactacca gggcttccag agcatgtaca cattcgtgac atctcatctg cctcctggct
     3301 tcaacgagta cgacttcgtg ccagagtctt tcgacaggga caaaaccatt gccctgatca
     3361 tgaacagctc tgggtctacc ggcctgccta agggcgtggc cctgcctcat cgcaccgcct
     3421 gtgtgcgctt ctctcacgcc cgcgacccta ttttcggcaa ccagatcatc cccgacaccg
     3481 ctattctgag cgtggtgcca ttccaccacg gcttcggcat gttcaccacc ctgggctacc
     3541 tgatttgcgg ctttcgggtg gtgctgatgt accgcttcga ggaggagctg ttcctgcgca
     3601 gcctgcaaga ctacaaaatt cagtctgccc tgctggtgcc aaccctgttc agcttcttcg
     3661 ctaagagcac cctgatcgac aagtacgacc tgtctaacct gcacgagatt gcctctggcg
     3721 gcgccccact gtctaaggag gtgggcgaag ccgtggccaa gcgctttcat ctgccaggca
     3781 tccgccaggg ctacggcctg accgagacaa ccagcgccat tctgattacc ccagagggcg
     3841 acgacaagcc tggcgccgtg ggcaaggtgg tgccattctt cgaggccaag gtggtggacc
     3901 tggacaccgg caagaccctg ggagtgaacc agcgcggcga gctgtgtgtg cgcggcccta
     3961 tgattatgtc cggctacgtg aataaccctg aggccacaaa cgccctgatc gacaaggacg
     4021 gctggctgca ctctggcgac attgcctact gggacgagga cgagcacttc ttcatcgtgg
     4081 accgcctgaa gtctctgatc aagtacaagg gctaccaggt ggccccagcc gagctggagt
     4141 ctatcctgct gcagcaccct aacattttcg acgccggagt ggccggcctg cccgacgacg
     4201 atgccggcga gctgcctgcc gccgtcgtcg tgctggaaca cggcaagacc atgaccgaga
     4261 aggagatcgt ggactatgtg gccagccagg tgacaaccgc caagaagctg cgcggcggag
     4321 tggtgttcgt ggacgaggtg cccaagggcc tgaccggcaa gctggacgcc cgcaagatcc
     4381 gcgagatcct gatcaaggct aagaaaggcg gcaagatcgc cgtgtaatgc ggccgcttcg
     4441 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     4501 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     4561 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     4621 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc
     4681 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4741 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4801 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4861 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4921 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4981 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5041 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5101 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5161 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5221 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5281 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5341 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5401 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5461 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5521 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5581 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5641 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5701 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5761 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5821 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5881 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5941 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6001 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6061 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6121 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6181 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6241 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6301 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6361 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6421 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6481 tgctcacatg gctcgaca
//
