LOCUS       sequence-315875-        3115 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for: ratiometric pHluorin (mitchondrial).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3115)
  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 3115)
  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: pMOS018E: Entry vector for: ratiometric pHluor...
FEATURES             Location/Qualifiers
     source          1..3115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             495..569
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             603..1316
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Miesenbock et al., 1998)"
                     /label=ratiometric pHluorin
                     /note="responds to a drop in pH with a reversible decrease 
                     in excitation at 395 nm and a corresponding increase in 
                     excitation at 475 nm"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKDDGNILGHKLEYNYNEHLVYIMADKQKNGTKAI
                     FQVHHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLHTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(630..658)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1253..1273
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    complement(1344..1443)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1555..1574)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1566..2375
                     /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(1633..1652)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2465..3053
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2954..2973
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggctggc gccggaacca attcagtcga
      481 ctggatccgc caccatgtcc gtcctgacgc cgctgctgct gcggggcttg acaggctcgg
      541 cccggcggct cccagtgccg cgcgccaaga tccattcgtt gggtgacccg ccagtcgcta
      601 gcagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg
      661 atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa
      721 aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg ccaacacttg
      781 tcactacttt ctcttatggt gttcaatgct tttcaagata cccagatcat atgaaacggc
      841 atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca
      901 aagatgacgg gaactacaag acacgtgctg aagtcaagtt tgaaggtgat acccttgtta
      961 atagaatcga gttaaaaggt attgatttta aagatgatgg aaacattctt ggacacaaat
     1021 tggaatacaa ctataacgag cacttggtgt acatcatggc agacaaacaa aagaatggta
     1081 ccaaagctat ctttcaagtt caccacaaca ttgaagatgg aggcgttcaa ctagcagacc
     1141 attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc
     1201 tgcacacaca atctgccctt tcgaaagatc ccaacgaaaa gagagaccac atggtccttc
     1261 ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac aaatgaattc
     1321 gcggccgcac tcgagatatc tagacccagc tttcttgtac aaagttggca ttataagaaa
     1381 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     1441 ttgccatcca gctgcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     1501 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     1561 gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg
     1621 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     1681 atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     1741 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     1801 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     1861 tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     1921 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     1981 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     2041 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     2101 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     2161 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     2221 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     2281 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     2341 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac
     2401 attattcaga ttgggccccg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2461 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2521 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2581 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     2641 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2701 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2761 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2821 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2881 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2941 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3001 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3061 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
