LOCUS       sequence-352606-        6600 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cytosolic firefly luciferase reporter previously described in 
            Shcherbakov et al. 2019.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6600)
  AUTHORS   Shcherbakov D, Teo Y, Boukari H, Cortes-Sanchon A, Mantovani M, 
            Osinnii I, Moore J, Juskeviciene R, Brilkova M, Duscha S, Kumar HS, 
            Laczko E, Rehrauer H, Westhof E, Akbergenov R, Bottger EC
  TITLE     Ribosomal mistranslation leads to silencing of the unfolded protein 
            response and increased mitochondrial biogenesis.
  JOURNAL   Commun Biol. 2019 Oct 17;2:381. doi: 10.1038/s42003-019-0626-9. 
            eCollection 2019.
  PUBMED    31637312
REFERENCE   2  (bases 1 to 6600)
  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: "Commun 
            Biol."; date: "2019-10-17"; volume: "2:381. doi"; pages: " 
            10.1038/s42003-019-0626-9. eCollection 2019"
COMMENT     Sequence Label: phFluc-WT
FEATURES             Location/Qualifiers
     source          1..6600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..43
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             142..1179
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             1158..1166
                     /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    1203..1251
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(1407..1424)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1578..2166)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1658..1677)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2366..3157,3158..3226))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    3331..3379
                     /note="synthetic polyadenylation signal"
     misc_feature    3393..3484
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3433..3452
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     enhancer        3635..4014
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4015..4218
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4169..4189
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4215..4239
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4337..4346
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4343..5995
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    6039..6160
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6050..6069)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6104..6123
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        join(6354..6600,1..111)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6354..6545
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6377..6397)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6562..6600,1..97)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
       61 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcgat
      121 tcttctgaca ctagcgccac catgaagaag cccgaactca ccgctaccag cgttgaaaaa
      181 tttctcatcg agaagttcga cagtgtgagc gacctgatgc agttgtcgga gggcgaagag
      241 agccgagcct tcagcttcga tgtcggcgga cgcggctatg tactgcgggt gaatagctgc
      301 gctgatggct tctacaaaga ccgctacgtg taccgccact tcgccagcgc tgcactaccc
      361 atccccgaag tgttggacat cggcgagttc agcgagagcc tgacatactg catcagtaga
      421 cgcgcccaag gcgttactct ccaagacctc cccgaaacag agctgcctgc tgtgttacag
      481 cctgtcgccg aagctatgga tgctattgcc gccgccgacc tcagtcaaac cagcggcttc
      541 ggcccattcg ggccccaagg catcggccag tacacaacct ggcgggattt catttgcgcc
      601 attgctgatc cccatgtcta ccactggcag accgtgatgg acgacaccgt gtccgccagc
      661 gtagctcaag ccctggacga actgatgctg tgggccgaag actgtcccga ggtgcgccac
      721 ctcgtccatg ccgacttcgg cagcaacaac gtcctgaccg acaacggccg catcaccgcc
      781 gtaatcgact ggtccgaagc tatgttcggg gacagtcagt acgaggtggc caacatcttc
      841 ttctggcggc cctggctggc ttgcatggag cagcagactc gctacttcga gcgccggcat
      901 cccgagctgg ccggcagccc tcgtctgcga gcctacatgc tgcgcatcgg cctggatcag
      961 ctctaccaga gcctcgtgga cggcaacttc gacgatgctg cctgggctca aggccgctgc
     1021 gatgccatcg tccgcagcgg ggccggcacc gtcggtcgca cacaaatcgc tcgccggagc
     1081 gcagccgtat ggaccgacgg ctgcgtcgag gtgctggccg acagcggcaa ccgccggccc
     1141 agtacacgac cgcgcgctaa ggaggtaggt cgagtttaaa ctctagaacc ggtcatggcc
     1201 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gttcgaacta
     1261 gatgctgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc
     1321 ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac
     1381 aggtgccggc agcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1441 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     1501 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     1561 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     1621 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     1681 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     1741 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     1801 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     1861 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     1921 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     1981 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2041 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2101 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2161 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2221 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2281 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagcggc
     2341 cgcaaatgct aaaccactgc agtggttacc agtgcttgat cagtgaggca ccgatctcag
     2401 cgatctgcct atttcgttcg tccatagtgg cctgactccc cgtcgtgtag atcactacga
     2461 ttcgtgaggg cttaccatca ggccccagcg cagcaatgat gccgcgagag ccgcgttcac
     2521 cggcccccga tttgtcagca atgaaccagc cagcagggag ggccgagcga agaagtggtc
     2581 ctgctacttt gtccgcctcc atccagtcta tgagctgctg tcgtgatgct agagtaagaa
     2641 gttcgccagt gagtagtttc cgaagagttg tggccattgc tactggcatc gtggtatcac
     2701 gctcgtcgtt cggtatggct tcgttcaact ctggttccca gcggtcaagc cgggtcacat
     2761 gatcacccat attatgaaga aatgcagtca gctccttagg gcctccgatc gttgtcagaa
     2821 gtaagttggc cgcggtgttg tcgctcatgg taatggcagc actacacaat tctcttaccg
     2881 tcatgccatc cgtaagatgc ttttccgtga ccggcgagta ctcaaccaag tcgttttgtg
     2941 agtagtgtat acggcgacca agctgctctt gcccggcgtc tatacgggac aacaccgcgc
     3001 cacatagcag tactttgaaa gtgctcatca tcgggaatcg ttcttcgggg cggaaagact
     3061 caaggatctt gccgctattg agatccagtt cgatatagcc cactcttgca cccagttgat
     3121 cttcagcatc ttttactttc accagcgttt cggggtgtgc aaaaacaggc aagcaaaatg
     3181 ccgcaaagaa gggaatgagt gcgacacgaa aatgttggat gctcatactc gtcctttttc
     3241 aatattattg aagcatttat cagggttact agtacgtctc tcaaggataa gtaagtaata
     3301 ttaaggtacg ggaggtattg gacaggccgc aataaaatat ctttattttc attacatctg
     3361 tgtgttggtt ttttgtgtga atcgatagta ctaacatacg ctctccatca aaacaaaacg
     3421 aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt
     3481 ctctggccta actggcctca atattggcca ttagccatat tattcattgg ttatatagca
     3541 taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt
     3601 tatattggct catgtccaat atgaccgcca tgttggcatt gattattgac tagttattaa
     3661 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     3721 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     3781 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     3841 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc
     3901 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     3961 cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     4021 cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     4081 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     4141 aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt acggtgggag
     4201 gtctatataa gcagagctcg tttagtgaac cgtcagatca ctagaagctt tattgcggta
     4261 gtttatcaca gttaaattgc taacgcagtc agtgggcctc ggcggccaag cttggcaatc
     4321 cggtactgtt ggtaaagcca ccatggaaga tgccaaaaac attaagaagg gcccagcgcc
     4381 attctaccca ctcgaagacg ggaccgccgg cgagcagctg cacaaagcca tgaagcgcta
     4441 cgccctggtg cccggcacca tcgcctttac cgacgcacat atcgaggtgg acattaccta
     4501 cgccgagtac ttcgagatga gcgttcggct ggcagaagct atgaagcgct atgggctgaa
     4561 tacaaaccat cggatcgtgg tgtgcagcga gaatagcttg cagttcttca tgcccgtgtt
     4621 gggtgccctg ttcatcggtg tggctgtggc cccagctaac gacatctaca acgagcgcga
     4681 gctgctgaac agcatgggca tcagccagcc caccgtcgta ttcgtgagca agaaagggct
     4741 gcaaaagatc ctcaacgtgc aaaagaagct accgatcata caaaagatca tcatcatgga
     4801 tagcaagacc gactaccagg gcttccaaag catgtacacc ttcgtgactt cccatttgcc
     4861 acccggcttc aacgagtacg acttcgtgcc cgagagcttc gaccgggaca aaaccatcgc
     4921 cctgatcatg aacagtagtg gcagtaccgg attgcccaag ggcgtagccc taccgcaccg
     4981 caccgcttgt gtccgattca gtcatgcccg cgaccccatc ttcggcaacc agatcatccc
     5041 cgacaccgct atcctcagcg tggtgccatt tcaccacggc ttcggcatgt tcaccacgct
     5101 gggctacttg atctgcggct ttcgggtcgt gctcatgtac cgcttcgagg aggagctatt
     5161 cttgcgcagc ttgcaagact ataagattca atctgccctg ctggtgccca cactatttag
     5221 cttcttcgct aagagcactc tcatcgacaa gtacgaccta agcaacttgc acgagatcgc
     5281 cagcggcggg gcgccgctca gcaaggaggt aggtgaggcc gtggccaaac gcttccacct
     5341 accaggcatc cgccagggct acggcctgac agaaacaacc agcgccattc tgatcacccc
     5401 cgaaggggac gacaagcctg gcgcagtagg caaggtggtg cccttcttcg aggctaaggt
     5461 ggtggacttg gacaccggta agacactggg tgtgaaccag cgcggcgagc tgtgcgtccg
     5521 tggccccatg atcatgagcg gctacgttaa caaccccgag gctacaaacg ctctcatcga
     5581 caaggacggc tggctgcaca gcggcgacat cgcctactgg gacgaggacg agcacttctt
     5641 catcgtggac cggctgaaga gcctgatcaa atacaagggc taccaggtag ccccagccga
     5701 actggagagc atcctgctgc aacaccccaa catcttcgac gccggggtcg ccggcctgcc
     5761 cgacgacgat gccggcgagc tgcccgccgc agtcgtcgtg ctggaacacg gtaaaaccat
     5821 gaccgagaag gagatcgtgg actatgtggc cagccaggtt acaaccgcca agaagctgcg
     5881 cggtggtgtt gtgttcgtgg acgaggtgcc taaaggactg accggcaagt tggacgcccg
     5941 caagatccgc gagattctca ttaaggccaa gaagggcggc aagatcgccg tgtaataatt
     6001 ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg
     6061 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     6121 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     6181 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     6241 caaatgtggt aaaatcgata aggatccgtt tgcgtattgg gcgctcttcc gctgatctgc
     6301 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttagctac cttctgaggc
     6361 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     6421 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     6481 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     6541 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
//
