LOCUS       sequence-352607-        6758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase reporter fused to a mitochondria signal sequence 
            for mitochondrial targeting.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6758)
  AUTHORS   Cortes Sanchon A, Santhosh Kumar H, Mantovani M, Osinnii I, Mateos 
            JM, Kaech A, Shcherbakov D, Akbergenov R, Bottger EC
  TITLE     ER-misfolded proteins become sequestered with mitochondria and 
            impair mitochondrial function.
  JOURNAL   Commun Biol. 2021 Dec 2;4(1):1350. doi: 10.1038/s42003-021-02873-w.
  PUBMED    34857875
REFERENCE   2  (bases 1 to 6758)
  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: "2021-12-2"; volume: "4(1):1350. doi"; pages: " 
            10.1038/s42003-021-02873-w"
COMMENT     Sequence Label: pMTS-hFluc
FEATURES             Location/Qualifiers
     source          1..6758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             394..402
                     /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    439..487
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(643..660)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(814..1402)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(894..913)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1602..2393,2394..2462))
                     /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    2567..2615
                     /note="synthetic polyadenylation signal"
     misc_feature    2629..2720
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2669..2688
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     enhancer        2871..3250
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3251..3454
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3405..3425
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3451..3475
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3740..5389
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="EDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAH
                     IEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA
                     NDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM
                     YTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHAR
                     DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKI
                     QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG
                     LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG
                     YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL
                     QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF
                     VDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    5433..5554
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5444..5463)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5498..5517
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5748..6105
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5748..5939
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5771..5791)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5956..6091
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6018..6037
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             join(6136..6758,1..415)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
ORIGIN
        1 cggccgcatc accgccgtaa tcgactggtc cgaagctatg ttcggggaca gtcagtacga
       61 ggtggccaac atcttcttct ggcggccctg gctggcttgc atggagcagc agactcgcta
      121 cttcgagcgc cggcatcccg agctggccgg cagccctcgt ctgcgagcct acatgctgcg
      181 catcggcctg gatcagctct accagagcct cgtggacggc aacttcgacg atgctgcctg
      241 ggctcaaggc cgctgcgatg ccatcgtccg cagcggggcc ggcaccgtcg gtcgcacaca
      301 aatcgctcgc cggagcgcag ccgtatggac cgacggctgc gtcgaggtgc tggccgacag
      361 cggcaaccgc cggcccagta cacgaccgcg cgctaaggag gtaggtcgag tttaaactct
      421 agaaccggtc atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt
      481 ttgtgtgttc gaactagatg ctgtcgaccg atgcccttga gagccttcaa cccagtcagc
      541 tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc
      601 atgcaactcg taggacaggt gccggcagcg ctcttccgct tcctcgctca ctgactcgct
      661 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
      721 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
      781 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
      841 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
      901 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      961 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     1021 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     1081 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     1141 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     1201 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     1261 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     1321 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     1381 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     1441 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     1501 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     1561 ttggtctgac agcggccgca aatgctaaac cactgcagtg gttaccagtg cttgatcagt
     1621 gaggcaccga tctcagcgat ctgcctattt cgttcgtcca tagtggcctg actccccgtc
     1681 gtgtagatca ctacgattcg tgagggctta ccatcaggcc ccagcgcagc aatgatgccg
     1741 cgagagccgc gttcaccggc ccccgatttg tcagcaatga accagccagc agggagggcc
     1801 gagcgaagaa gtggtcctgc tactttgtcc gcctccatcc agtctatgag ctgctgtcgt
     1861 gatgctagag taagaagttc gccagtgagt agtttccgaa gagttgtggc cattgctact
     1921 ggcatcgtgg tatcacgctc gtcgttcggt atggcttcgt tcaactctgg ttcccagcgg
     1981 tcaagccggg tcacatgatc acccatatta tgaagaaatg cagtcagctc cttagggcct
     2041 ccgatcgttg tcagaagtaa gttggccgcg gtgttgtcgc tcatggtaat ggcagcacta
     2101 cacaattctc ttaccgtcat gccatccgta agatgctttt ccgtgaccgg cgagtactca
     2161 accaagtcgt tttgtgagta gtgtatacgg cgaccaagct gctcttgccc ggcgtctata
     2221 cgggacaaca ccgcgccaca tagcagtact ttgaaagtgc tcatcatcgg gaatcgttct
     2281 tcggggcgga aagactcaag gatcttgccg ctattgagat ccagttcgat atagcccact
     2341 cttgcaccca gttgatcttc agcatctttt actttcacca gcgtttcggg gtgtgcaaaa
     2401 acaggcaagc aaaatgccgc aaagaaggga atgagtgcga cacgaaaatg ttggatgctc
     2461 atactcgtcc tttttcaata ttattgaagc atttatcagg gttactagta cgtctctcaa
     2521 ggataagtaa gtaatattaa ggtacgggag gtattggaca ggccgcaata aaatatcttt
     2581 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct
     2641 ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc
     2701 aggtgccaga acatttctct ggcctaactg gcctcaatat tggccattag ccatattatt
     2761 cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata
     2821 tcataatatg tacatttata ttggctcatg tccaatatga ccgccatgtt ggcattgatt
     2881 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     2941 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     3001 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     3061 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     3121 tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     3181 ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     3241 tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc
     3301 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     3361 tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag
     3421 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcactag
     3481 aagcttaacc atggcctcca ctcgtgtcct cgcctctcgc ctggcctccc ggatggctgc
     3541 ttccgccaag gttgcccgcc ctgctgtccg cgttgctcag gtcagcaagc gcaccatcca
     3601 gactggctcc cccctccaga ccctcaagcg cacccagatg acctccatcg tcaacgccac
     3661 cacccgccag gctttccaga agcgcgccta ctcttcctgg aattcagctt ggcaatccgg
     3721 tactgttggt aaagccaccg aagatgccaa aaacattaag aagggcccag cgccattcta
     3781 cccactcgaa gacgggaccg ccggcgagca gctgcacaaa gccatgaagc gctacgccct
     3841 ggtgcccggc accatcgcct ttaccgacgc acatatcgag gtggacatta cctacgccga
     3901 gtacttcgag atgagcgttc ggctggcaga agctatgaag cgctatgggc tgaatacaaa
     3961 ccatcggatc gtggtgtgca gcgagaatag cttgcagttc ttcatgcccg tgttgggtgc
     4021 cctgttcatc ggtgtggctg tggccccagc taacgacatc tacaacgagc gcgagctgct
     4081 gaacagcatg ggcatcagcc agcccaccgt cgtattcgtg agcaagaaag ggctgcaaaa
     4141 gatcctcaac gtgcaaaaga agctaccgat catacaaaag atcatcatca tggatagcaa
     4201 gaccgactac cagggcttcc aaagcatgta caccttcgtg acttcccatt tgccacccgg
     4261 cttcaacgag tacgacttcg tgcccgagag cttcgaccgg gacaaaacca tcgccctgat
     4321 catgaacagt agtggcagta ccggattgcc caagggcgta gccctaccgc accgcaccgc
     4381 ttgtgtccga ttcagtcatg cccgcgaccc catcttcggc aaccagatca tccccgacac
     4441 cgctatcctc agcgtggtgc catttcacca cggcttcggc atgttcacca cgctgggcta
     4501 cttgatctgc ggctttcggg tcgtgctcat gtaccgcttc gaggaggagc tattcttgcg
     4561 cagcttgcaa gactataaga ttcaatctgc cctgctggtg cccacactat ttagcttctt
     4621 cgctaagagc actctcatcg acaagtacga cctaagcaac ttgcacgaga tcgccagcgg
     4681 cggggcgccg ctcagcaagg aggtaggtga ggccgtggcc aaacgcttcc acctaccagg
     4741 catccgccag ggctacggcc tgacagaaac aaccagcgcc attctgatca cccccgaagg
     4801 ggacgacaag cctggcgcag taggcaaggt ggtgcccttc ttcgaggcta aggtggtgga
     4861 cttggacacc ggtaagacac tgggtgtgaa ccagcgcggc gagctgtgcg tccgtggccc
     4921 catgatcatg agcggctacg ttaacaaccc cgaggctaca aacgctctca tcgacaagga
     4981 cggctggctg cacagcggcg acatcgccta ctgggacgag gacgagcact tcttcatcgt
     5041 ggaccggctg aagagcctga tcaaatacaa gggctaccag gtagccccag ccgaactgga
     5101 gagcatcctg ctgcaacacc ccaacatctt cgacgccggg gtcgccggcc tgcccgacga
     5161 cgatgccggc gagctgcccg ccgcagtcgt cgtgctggaa cacggtaaaa ccatgaccga
     5221 gaaggagatc gtggactatg tggccagcca ggttacaacc gccaagaagc tgcgcggtgg
     5281 tgttgtgttc gtggacgagg tgcctaaagg actgaccggc aagttggacg cccgcaagat
     5341 ccgcgagatt ctcattaagg ccaagaaggg cggcaagatc gccgtgtaat aattctagag
     5401 tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     5461 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     5521 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     5581 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     5641 tggtaaaatc gataaggatc cgtttgcgta ttgggcgctc ttccgctgat ctgcgcagca
     5701 ccatggcctg aaataacctc tgaaagagga acttggttag ctaccttctg aggcggaaag
     5761 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     5821 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     5881 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     5941 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     6001 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     6061 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cgattcttct
     6121 gacactagcg ccaccatgaa gaagcccgaa ctcaccgcta ccagcgttga aaaatttctc
     6181 atcgagaagt tcgacagtgt gagcgacctg atgcagttgt cggagggcga agagagccga
     6241 gccttcagct tcgatgtcgg cggacgcggc tatgtactgc gggtgaatag ctgcgctgat
     6301 ggcttctaca aagaccgcta cgtgtaccgc cacttcgcca gcgctgcact acccatcccc
     6361 gaagtgttgg acatcggcga gttcagcgag agcctgacat actgcatcag tagacgcgcc
     6421 caaggcgtta ctctccaaga cctccccgaa acagagctgc ctgctgtgtt acagcctgtc
     6481 gccgaagcta tggatgctat tgccgccgcc gacctcagtc aaaccagcgg cttcggccca
     6541 ttcgggcccc aaggcatcgg ccagtacaca acctggcggg atttcatttg cgccattgct
     6601 gatccccatg tctaccactg gcagaccgtg atggacgaca ccgtgtccgc cagcgtagct
     6661 caagccctgg acgaactgat gctgtgggcc gaagactgtc ccgaggtgcg ccacctcgtc
     6721 catgccgact tcggcagcaa caacgtcctg accgacaa
//
