LOCUS       sequence-352620-        6908 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase reporter fused to a mitochondria signal sequence 
            for mitochondrial targeting with an HA tag and the eleventh 
            beta-strand of GFP added to the C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6908)
  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 6908)
  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-HA-GFP11
FEATURES             Location/Qualifiers
     source          1..6908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..34
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             133..1170
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             1149..1157
                     /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    1194..1242
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(1398..1415)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1569..2157)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1649..1668)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2357..3148,3149..3217))
                     /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    3322..3370
                     /note="synthetic polyadenylation signal"
     misc_feature    3384..3475
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3424..3443
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     enhancer        3626..4005
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4006..4209
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4160..4180
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4206..4230
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4495..6141
                     /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"
     CDS             6178..6204
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6244..6291
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     polyA_signal    6338..6459
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6349..6368)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6403..6422
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        join(6653..6908,1..102)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6653..6844
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6676..6696)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6861..6908,1..88)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
       61 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctcga ttcttctgac
      121 actagcgcca ccatgaagaa gcccgaactc accgctacca gcgttgaaaa atttctcatc
      181 gagaagttcg acagtgtgag cgacctgatg cagttgtcgg agggcgaaga gagccgagcc
      241 ttcagcttcg atgtcggcgg acgcggctat gtactgcggg tgaatagctg cgctgatggc
      301 ttctacaaag accgctacgt gtaccgccac ttcgccagcg ctgcactacc catccccgaa
      361 gtgttggaca tcggcgagtt cagcgagagc ctgacatact gcatcagtag acgcgcccaa
      421 ggcgttactc tccaagacct ccccgaaaca gagctgcctg ctgtgttaca gcctgtcgcc
      481 gaagctatgg atgctattgc cgccgccgac ctcagtcaaa ccagcggctt cggcccattc
      541 gggccccaag gcatcggcca gtacacaacc tggcgggatt tcatttgcgc cattgctgat
      601 ccccatgtct accactggca gaccgtgatg gacgacaccg tgtccgccag cgtagctcaa
      661 gccctggacg aactgatgct gtgggccgaa gactgtcccg aggtgcgcca cctcgtccat
      721 gccgacttcg gcagcaacaa cgtcctgacc gacaacggcc gcatcaccgc cgtaatcgac
      781 tggtccgaag ctatgttcgg ggacagtcag tacgaggtgg ccaacatctt cttctggcgg
      841 ccctggctgg cttgcatgga gcagcagact cgctacttcg agcgccggca tcccgagctg
      901 gccggcagcc ctcgtctgcg agcctacatg ctgcgcatcg gcctggatca gctctaccag
      961 agcctcgtgg acggcaactt cgacgatgct gcctgggctc aaggccgctg cgatgccatc
     1021 gtccgcagcg gggccggcac cgtcggtcgc acacaaatcg ctcgccggag cgcagccgta
     1081 tggaccgacg gctgcgtcga ggtgctggcc gacagcggca accgccggcc cagtacacga
     1141 ccgcgcgcta aggaggtagg tcgagtttaa actctagaac cggtcatggc cgcaataaaa
     1201 tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgttcgaact agatgctgtc
     1261 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     1321 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     1381 cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1441 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1501 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1561 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1621 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1681 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1741 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1801 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1861 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     1921 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     1981 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2041 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2101 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2161 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2221 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2281 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagcgg ccgcaaatgc
     2341 taaaccactg cagtggttac cagtgcttga tcagtgaggc accgatctca gcgatctgcc
     2401 tatttcgttc gtccatagtg gcctgactcc ccgtcgtgta gatcactacg attcgtgagg
     2461 gcttaccatc aggccccagc gcagcaatga tgccgcgaga gccgcgttca ccggcccccg
     2521 atttgtcagc aatgaaccag ccagcaggga gggccgagcg aagaagtggt cctgctactt
     2581 tgtccgcctc catccagtct atgagctgct gtcgtgatgc tagagtaaga agttcgccag
     2641 tgagtagttt ccgaagagtt gtggccattg ctactggcat cgtggtatca cgctcgtcgt
     2701 tcggtatggc ttcgttcaac tctggttccc agcggtcaag ccgggtcaca tgatcaccca
     2761 tattatgaag aaatgcagtc agctccttag ggcctccgat cgttgtcaga agtaagttgg
     2821 ccgcggtgtt gtcgctcatg gtaatggcag cactacacaa ttctcttacc gtcatgccat
     2881 ccgtaagatg cttttccgtg accggcgagt actcaaccaa gtcgttttgt gagtagtgta
     2941 tacggcgacc aagctgctct tgcccggcgt ctatacggga caacaccgcg ccacatagca
     3001 gtactttgaa agtgctcatc atcgggaatc gttcttcggg gcggaaagac tcaaggatct
     3061 tgccgctatt gagatccagt tcgatatagc ccactcttgc acccagttga tcttcagcat
     3121 cttttacttt caccagcgtt tcggggtgtg caaaaacagg caagcaaaat gccgcaaaga
     3181 agggaatgag tgcgacacga aaatgttgga tgctcatact cgtccttttt caatattatt
     3241 gaagcattta tcagggttac tagtacgtct ctcaaggata agtaagtaat attaaggtac
     3301 gggaggtatt ggacaggccg caataaaata tctttatttt cattacatct gtgtgttggt
     3361 tttttgtgtg aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac
     3421 aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctggcct
     3481 aactggcctc aatattggcc attagccata ttattcattg gttatatagc ataaatcaat
     3541 attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc
     3601 tcatgtccaa tatgaccgcc atgttggcat tgattattga ctagttatta atagtaatca
     3661 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     3721 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     3781 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     3841 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac
     3901 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt
     3961 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     4021 cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     4081 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     4141 aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     4201 agcagagctc gtttagtgaa ccgtcagatc actagaagct taaccatggc ctccactcgt
     4261 gtcctcgcct ctcgcctggc ctcccggatg gctgcttccg ccaaggttgc ccgccctgct
     4321 gtccgcgttg ctcaggtcag caagcgcacc atccagactg gctcccccct ccagaccctc
     4381 aagcgcaccc agatgacctc catcgtcaac gccaccaccc gccaggcttt ccagaagcgc
     4441 gcctactctt cctggaattc agcttggcaa tccggtactg ttggtaaagc caccgaagat
     4501 gccaaaaaca ttaagaaggg cccagcgcca ttctacccac tcgaagacgg gaccgccggc
     4561 gagcagctgc acaaagccat gaagcgctac gccctggtgc ccggcaccat cgcctttacc
     4621 gacgcacata tcgaggtgga cattacctac gccgagtact tcgagatgag cgttcggctg
     4681 gcagaagcta tgaagcgcta tgggctgaat acaaaccatc ggatcgtggt gtgcagcgag
     4741 aatagcttgc agttcttcat gcccgtgttg ggtgccctgt tcatcggtgt ggctgtggcc
     4801 ccagctaacg acatctacaa cgagcgcgag ctgctgaaca gcatgggcat cagccagccc
     4861 accgtcgtat tcgtgagcaa gaaagggctg caaaagatcc tcaacgtgca aaagaagcta
     4921 ccgatcatac aaaagatcat catcatggat agcaagaccg actaccaggg cttccaaagc
     4981 atgtacacct tcgtgacttc ccatttgcca cccggcttca acgagtacga cttcgtgccc
     5041 gagagcttcg accgggacaa aaccatcgcc ctgatcatga acagtagtgg cagtaccgga
     5101 ttgcccaagg gcgtagccct accgcaccgc accgcttgtg tccgattcag tcatgcccgc
     5161 gaccccatct tcggcaacca gatcatcccc gacaccgcta tcctcagcgt ggtgccattt
     5221 caccacggct tcggcatgtt caccacgctg ggctacttga tctgcggctt tcgggtcgtg
     5281 ctcatgtacc gcttcgagga ggagctattc ttgcgcagct tgcaagacta taagattcaa
     5341 tctgccctgc tggtgcccac actatttagc ttcttcgcta agagcactct catcgacaag
     5401 tacgacctaa gcaacttgca cgagatcgcc agcggcgggg cgccgctcag caaggaggta
     5461 ggtgaggccg tggccaaacg cttccaccta ccaggcatcc gccagggcta cggcctgaca
     5521 gaaacaacca gcgccattct gatcaccccc gaaggggacg acaagcctgg cgcagtaggc
     5581 aaggtggtgc ccttcttcga ggctaaggtg gtggacttgg acaccggtaa gacactgggt
     5641 gtgaaccagc gcggcgagct gtgcgtccgt ggccccatga tcatgagcgg ctacgttaac
     5701 aaccccgagg ctacaaacgc tctcatcgac aaggacggct ggctgcacag cggcgacatc
     5761 gcctactggg acgaggacga gcacttcttc atcgtggacc ggctgaagag cctgatcaaa
     5821 tacaagggct accaggtagc cccagccgaa ctggagagca tcctgctgca acaccccaac
     5881 atcttcgacg ccggggtcgc cggcctgccc gacgacgatg ccggcgagct gcccgccgca
     5941 gtcgtcgtgc tggaacacgg taaaaccatg accgagaagg agatcgtgga ctatgtggcc
     6001 agccaggtta caaccgccaa gaagctgcgc ggtggtgttg tgttcgtgga cgaggtgcct
     6061 aaaggactga ccggcaagtt ggacgcccgc aagatccgcg agattctcat taaggccaag
     6121 aagggcggca agatcgccgt gatccccggg ggatcgggtg acggtccggt cgccacctac
     6181 ccatacgatg ttccagatta cgctggtcga cggatcggtg gtggttctgg aggcggttct
     6241 atgagagatc atatggtttt gcatgaatat gttaatgctg ctggtattac ttaataattc
     6301 tagagtcggg gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     6361 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     6421 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     6481 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     6541 aaatgtggta aaatcgataa ggatccgttt gcgtattggg cgctcttccg ctgatctgcg
     6601 cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttagctacc ttctgaggcg
     6661 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     6721 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     6781 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     6841 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     6901 cccatggc
//
