LOCUS       sequence-464900-        5506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Rluc-Fluc reporter gene containing 39xY.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5506)
  AUTHORS   Matsuura-Suzuki E, Shimazu T, Takahashi M, Kotoshiba K, Suzuki T, 
            Kashiwagi K, Sohtome Y, Akakabe M, Sodeoka M, Dohmae N, Ito T, 
            Shinkai Y, Iwasaki S
  TITLE     METTL18-mediated histidine methylation of RPL3 modulates translation
            elongation for proteostasis maintenance.
  JOURNAL   Elife. 2022 Jun 8;11. pii: 72780. doi: 10.7554/eLife.72780.
  PUBMED    35674491
REFERENCE   2  (bases 1 to 5506)
  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"; journalName: "Elife."; 
            date: "2022-06-8"; volume: "11. pii: 72780. doi"; pages: " 
            10.7554/eLife.72780"
COMMENT     Sequence Label: psiCHECK2-1_Rluc-Y39x-Fluc
FEATURES             Location/Qualifiers
     source          1..5506
                     /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"
     primer_bind     604..623
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             706..1638
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             1765..3417
                     /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             2724..2732
                     /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    3461..3582
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3472..3491)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3526..3545
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3715..3819
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3820..3888,3889..4680)
                     /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(4038..4057)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4851..5439
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5340..5359
                     /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 taggtttcct tcctcttttg gcaacatggc ttccaaggtg
      721 tacgaccccg agcaacgcaa acgcatgatc actgggcctc agtggtgggc tcgctgcaag
      781 caaatgaacg tgctggactc cttcatcaac tactatgatt ccgagaagca cgccgagaac
      841 gccgtgattt ttctgcatgg taacgctgcc tccagctacc tgtggaggca cgtcgtgcct
      901 cacatcgagc ccgtggctag atgcatcatc cctgatctga tcggaatggg taagtccggc
      961 aagagcggga atggctcata tcgcctcctg gatcactaca agtacctcac cgcttggttc
     1021 gagctgctga accttccaaa gaaaatcatc tttgtgggcc acgactgggg ggcttgtctg
     1081 gcctttcact actcctacga gcaccaagac aagatcaagg ccatcgtcca tgctgagagt
     1141 gtcgtggacg tgatcgagtc ctgggacgag tggcctgaca tcgaggagga tatcgccctg
     1201 atcaagagcg aagagggcga gaaaatggtg cttgagaata acttcttcgt cgagaccatg
     1261 ctcccaagca agatcatgcg gaaactggag cctgaggagt tcgctgccta cctggagcca
     1321 ttcaaggaga agggcgaggt tagacggcct accctctcct ggcctcgcga gatccctctc
     1381 gttaagggag gcaagcccga cgtcgtccag attgtccgca actacaacgc ctaccttcgg
     1441 gccagcgacg atctgcctaa gatgttcatc gagtccgacc ctgggttctt ttccaacgct
     1501 attgtcgagg gagctaagaa gttccctaac accgagttcg tgaaggtgaa gggcctccac
     1561 ttcagccagg aggacgctcc agatgaaatg ggtaagtaca tcaagagctt cgtggagcgc
     1621 gtgctgaaga acgagcagat gtcgacgtac tactactact actactacta ctactactac
     1681 tactactact actactacta ctactactac tactactact actactacta ctactactac
     1741 tactactact actactacta ctacatggcc gatgctaaga acattaagaa gggccctgct
     1801 cccttctacc ctctggagga tggcaccgct ggcgagcagc tgcacaaggc catgaagagg
     1861 tatgccctgg tgcctggcac cattgccttc accgatgccc acattgaggt ggacatcacc
     1921 tatgccgagt acttcgagat gtctgtgcgc ctggccgagg ccatgaagag gtacggcctg
     1981 aacaccaacc accgcatcgt ggtgtgctct gagaactctc tgcagttctt catgccagtg
     2041 ctgggcgccc tgttcatcgg agtggccgtg gcccctgcta acgacattta caacgagcgc
     2101 gagctgctga acagcatggg catttctcag cctaccgtgg tgttcgtgtc taagaagggc
     2161 ctgcagaaga tcctgaacgt gcagaagaag ctgcctatca tccagaagat catcatcatg
     2221 gactctaaga ccgactacca gggcttccag agcatgtaca cattcgtgac atctcatctg
     2281 cctcctggct tcaacgagta cgacttcgtg ccagagtctt tcgacaggga caaaaccatt
     2341 gccctgatca tgaacagctc tgggtctacc ggcctgccta agggcgtggc cctgcctcat
     2401 cgcaccgcct gtgtgcgctt ctctcacgcc cgcgacccta ttttcggcaa ccagatcatc
     2461 cccgacaccg ctattctgag cgtggtgcca ttccaccacg gcttcggcat gttcaccacc
     2521 ctgggctacc tgatttgcgg ctttcgggtg gtgctgatgt accgcttcga ggaggagctg
     2581 ttcctgcgca gcctgcaaga ctacaaaatt cagtctgccc tgctggtgcc aaccctgttc
     2641 agcttcttcg ctaagagcac cctgatcgac aagtacgacc tgtctaacct gcacgagatt
     2701 gcctctggcg gcgccccact gtctaaggag gtgggcgaag ccgtggccaa gcgctttcat
     2761 ctgccaggca tccgccaggg ctacggcctg accgagacaa ccagcgccat tctgattacc
     2821 ccagagggcg acgacaagcc tggcgccgtg ggcaaggtgg tgccattctt cgaggccaag
     2881 gtggtggacc tggacaccgg caagaccctg ggagtgaacc agcgcggcga gctgtgtgtg
     2941 cgcggcccta tgattatgtc cggctacgtg aataaccctg aggccacaaa cgccctgatc
     3001 gacaaggacg gctggctgca ctctggcgac attgcctact gggacgagga cgagcacttc
     3061 ttcatcgtgg accgcctgaa gtctctgatc aagtacaagg gctaccaggt ggccccagcc
     3121 gagctggagt ctatcctgct gcagcaccct aacattttcg acgccggagt ggccggcctg
     3181 cccgacgacg atgccggcga gctgcctgcc gccgtcgtcg tgctggaaca cggcaagacc
     3241 atgaccgaga aggagatcgt ggactatgtg gccagccagg tgacaaccgc caagaagctg
     3301 cgcggcggag tggtgttcgt ggacgaggtg cccaagggcc tgaccggcaa gctggacgcc
     3361 cgcaagatcc gcgagatcct gatcaaggct aagaaaggcg gcaagatcgc cgtgtaataa
     3421 ttctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt
     3481 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     3541 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     3601 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     3661 tacaaatgtg gtaaaatcga taaggatcca ggtggcactt ttcggggaaa tgtgcgcgga
     3721 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3781 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3841 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3901 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3961 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4021 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4081 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4141 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4201 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4261 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4321 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4381 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4441 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4501 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4561 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4621 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4681 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4741 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4801 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4861 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4921 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4981 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5041 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5101 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5161 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5221 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5281 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5341 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5401 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5461 ttacggttcc tggccttttg ctggcctttt gctcacatgg ctcgac
//
