LOCUS       sequence-325109-        3997 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for: mito-roGFP2-Orp1 H2O2 oxidation sensor 
            (mitochondrial).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3997)
  AUTHORS   Werley CA, Boccardo S, Rigamonti A, Hansson EM, Cohen AE
  TITLE     Multiplexed Optical Sensors in Arrayed Islands of Cells for 
            multimodal recordings of cellular physiology.
  JOURNAL   Nat Commun. 2020 Aug 4;11(1):3881. doi: 10.1038/s41467-020-17607-5.
  PUBMED    32753572
REFERENCE   2  (bases 1 to 3997)
  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: "Nat 
            Commun."; date: "2020-08-4"; volume: "11(1):3881. doi"; pages: " 
            10.1038/s41467-020-17607-5"
COMMENT     Sequence Label: pMOS015E: Entry vector for: mito-roGFP2-Orp1 H...
FEATURES             Location/Qualifiers
     source          1..3997
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             707..781
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             join(806..808,809..1519)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(848..869)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1109..1128)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1456..1477
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2122..2221)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2238..2257)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2239..2257)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2262..2278)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2262..2278)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2380..2399)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2391..3200
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2458..2477)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3347..3935
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3836..3855
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcttt aaaggaacca attcagtcga ctggatctgc ggccgcatgt ccgtcctgac
      721 gccgctgctg ctgcggggct tgacaggctc ggcccggcgg ctcccagtgc cgcgcgccaa
      781 gatccattcg ttgccgccgg gatccgtgag caagggcgag gagctgttca ccggggtggt
      841 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
      901 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctcca ccaccggcaa
      961 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     1021 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     1081 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     1141 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1201 ggacggcaac atcctggggc acaagctgga gtacaactac aactgccaca acgtctatat
     1261 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     1321 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1381 cgtgctgctg cccgacaacc actacctgag cacctgctcc gccctgagca aagaccccaa
     1441 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1501 catggacgag ctgtacaaga ctagtggtgg ttcaggtggt ggtggttcag gtggtggtgg
     1561 ttcaggtgga ggaggatcag gaggaggagg atcaggagga ggaggatcag gaggagaatt
     1621 cgatatctca gaattctata agctagcacc tgttgacaag aaaggccaac cattcccctt
     1681 cgaccaatta aagggaaaag tggtgcttat cgttaatgtt gcctccaaat gtggattcac
     1741 tcctcaatac aaagaactag aggccttgta caaacgttat aaggacgaag gatttaccat
     1801 catcgggttc ccatgcaacc agtttggcca ccaagaacct ggctctgatg aagaaattgc
     1861 ccagttctgc caactgaact atggcgtgac tttccccatt atgaaaaaaa ttgacgttaa
     1921 tggtggcaat gaggaccctg tttacaagtt tttgaagagc caaaaatccg gtatgttggg
     1981 cttgagaggt atcaaatgga attttgaaaa attcttagtc gataaaaagg gtaaagtgta
     2041 cgaaagatac tcttcactaa ccaaaccttc ttcgttgtcc gaaaccatcg aagaactttt
     2101 gaaagaggtg gaatagtcta gacccagctt tcttgtacaa agttggcatt ataagaaagc
     2161 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
     2221 gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc
     2281 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     2341 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
     2401 ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     2461 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga
     2521 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     2581 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     2641 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag
     2701 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     2761 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     2821 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     2881 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
     2941 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     3001 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     3061 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     3121 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     3181 tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta
     3241 cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt
     3301 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3361 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3421 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3481 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3541 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3601 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3661 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3721 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3781 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3841 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3901 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3961 ggttcctggc cttttgctgg ccttttgctc acatgtt
//
