LOCUS       sequence-465633-        5419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Rluc-Fluc reporter gene containing no Y.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5419)
  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 5419)
  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_Rluc-Y0x-Fluc
FEATURES             Location/Qualifiers
     source          1..5419
                     /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="MASKVYYPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             1678..3330
                     /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             2637..2645
                     /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    3374..3495
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3385..3404)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3439..3458
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3628..3732
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3733..3801,3802..4593)
                     /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(3951..3970)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4764..5352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5253..5272
                     /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 tactaccccg 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 gtcgacgggg gcccctagga gatcccccgg ggagctcatg
     1681 gccgatgcta agaacattaa gaagggccct gctcccttct accctctgga ggatggcacc
     1741 gctggcgagc agctgcacaa ggccatgaag aggtatgccc tggtgcctgg caccattgcc
     1801 ttcaccgatg cccacattga ggtggacatc acctatgccg agtacttcga gatgtctgtg
     1861 cgcctggccg aggccatgaa gaggtacggc ctgaacacca accaccgcat cgtggtgtgc
     1921 tctgagaact ctctgcagtt cttcatgcca gtgctgggcg ccctgttcat cggagtggcc
     1981 gtggcccctg ctaacgacat ttacaacgag cgcgagctgc tgaacagcat gggcatttct
     2041 cagcctaccg tggtgttcgt gtctaagaag ggcctgcaga agatcctgaa cgtgcagaag
     2101 aagctgccta tcatccagaa gatcatcatc atggactcta agaccgacta ccagggcttc
     2161 cagagcatgt acacattcgt gacatctcat ctgcctcctg gcttcaacga gtacgacttc
     2221 gtgccagagt ctttcgacag ggacaaaacc attgccctga tcatgaacag ctctgggtct
     2281 accggcctgc ctaagggcgt ggccctgcct catcgcaccg cctgtgtgcg cttctctcac
     2341 gcccgcgacc ctattttcgg caaccagatc atccccgaca ccgctattct gagcgtggtg
     2401 ccattccacc acggcttcgg catgttcacc accctgggct acctgatttg cggctttcgg
     2461 gtggtgctga tgtaccgctt cgaggaggag ctgttcctgc gcagcctgca agactacaaa
     2521 attcagtctg ccctgctggt gccaaccctg ttcagcttct tcgctaagag caccctgatc
     2581 gacaagtacg acctgtctaa cctgcacgag attgcctctg gcggcgcccc actgtctaag
     2641 gaggtgggcg aagccgtggc caagcgcttt catctgccag gcatccgcca gggctacggc
     2701 ctgaccgaga caaccagcgc cattctgatt accccagagg gcgacgacaa gcctggcgcc
     2761 gtgggcaagg tggtgccatt cttcgaggcc aaggtggtgg acctggacac cggcaagacc
     2821 ctgggagtga accagcgcgg cgagctgtgt gtgcgcggcc ctatgattat gtccggctac
     2881 gtgaataacc ctgaggccac aaacgccctg atcgacaagg acggctggct gcactctggc
     2941 gacattgcct actgggacga ggacgagcac ttcttcatcg tggaccgcct gaagtctctg
     3001 atcaagtaca agggctacca ggtggcccca gccgagctgg agtctatcct gctgcagcac
     3061 cctaacattt tcgacgccgg agtggccggc ctgcccgacg acgatgccgg cgagctgcct
     3121 gccgccgtcg tcgtgctgga acacggcaag accatgaccg agaaggagat cgtggactat
     3181 gtggccagcc aggtgacaac cgccaagaag ctgcgcggcg gagtggtgtt cgtggacgag
     3241 gtgcccaagg gcctgaccgg caagctggac gcccgcaaga tccgcgagat cctgatcaag
     3301 gctaagaaag gcggcaagat cgccgtgtaa taattctaga gtcggggcgg ccggccgctt
     3361 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     3421 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     3481 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     3541 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat
     3601 ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3661 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3721 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3781 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3841 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3901 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3961 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4021 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4081 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4141 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4201 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4261 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4321 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4381 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4441 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4501 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4561 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4621 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4681 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4741 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4801 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4861 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4921 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4981 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5041 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5101 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5161 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5221 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5281 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5341 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5401 tttgctcaca tggctcgac
//
