LOCUS       sequence-488289-        6226 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  encodes a pH-sensitive ratiometric GFP (pH-GFP), which allows for 
            noninvasive measurements of intrabacterial pH in live cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6226)
  AUTHORS   Vandal OH, Pierini LM, Schnappinger D, Nathan CF, Ehrt S
  TITLE     A membrane protein preserves intrabacterial pH in intraphagosomal 
            Mycobacterium tuberculosis.
  JOURNAL   Nat Med. 2008 Aug;14(8):849-54. doi: 10.1038/nm.1795. Epub 2008 Jul 
            20.
  PUBMED    18641659
REFERENCE   2  (bases 1 to 6226)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nm.1795"; 
            journalName: "Nat Med"; date: "2008-08"; volume: "14"; issue: "8"; 
            pages: "849-54"
COMMENT     Sequence Label: pUV15-pHGFP
FEATURES             Location/Qualifiers
     source          1..6226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      20..47
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             404..1120
                     /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="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKDDGNILGHKLEYNYNEHLVYIMADKQKNGTKA
                     IFQVHHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLHTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(434..462)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1057..1077
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1181..1197)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1182..1198)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     rep_origin      5397..5985
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5886..5905
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6139..6156
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 actagttgat tagctaagca gaaggccatc ctgacggatg gcctttttgc gtttaatact
       61 gtttaaactc tagaaatatt ggatcgtcgg caccgtcacg gccgtgggag gcggcacgat
      121 ccgcgacgtg atgatcggcc gcatccccac ggtgctgcgc agtgagctct acgccatccc
      181 ggcgttgatc tgtgcgttcg cacgcacagg cccggtgtga gaagggtctc tgacgagcgg
      241 gagaacccac ccggggtggg cgagtttgtc ctgcgtgtgc tcggtcgagt aggctctggg
      301 agtacccgtg tgtacgacca gcacggcata catcatttcg acgccgagag attcgccgcc
      361 cgaaatgagc acgatccgca tgcttaatta aggaggcacc ggtatgagta aaggagaaga
      421 acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta atgggcacaa
      481 attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta cccttaaatt
      541 tatttgcact actggaaaac tacctgttcc atggccaaca cttgtcacta ctttctctta
      601 tggtgttcaa tgcttttcaa gatacccaga tcatatgaaa cggcatgact ttttcaagag
      661 tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg acgggaacta
      721 caagacacgt gctgaagtca agtttgaagg tgataccctt gttaatagaa tcgagttaaa
      781 aggtattgat tttaaagatg atggaaacat tcttggacac aaattggaat acaactataa
      841 cgagcacttg gtgtacatca tggcagacaa acaaaagaat ggtaccaaag ctatctttca
      901 agttcaccac aacattgaag atggaggcgt tcaactagca gaccattatc aacaaaatac
      961 tccaattggc gatggccctg tccttttacc agacaaccat tacctgcaca cacaatctgc
     1021 cctttcgaaa gatcccaacg aaaagagaga ccacatggtc cttcttgagt ttgtaacagc
     1081 tgctgggatt acacatggca tggatgaact atacaaataa ggtgatatca agctttagct
     1141 aattaattgg ggaccctaga ggtccccttt tttattttat cgataccgtc gacctcgagg
     1201 gggggccccc ggggatcatc gagccgagaa cgttatcgaa gttggtcatg tgtaatcccc
     1261 tcgtttgaac tttggattaa gcgtagatac acccttggac aagccagttg gattcggaga
     1321 caagcaaatt cagccttaaa aagggcgagg ccctgcggtg gtggaacacc gcagggcctc
     1381 taaccgctcg acgcgctgca ccaaccagcc cgcgaacggc tggcagccag cgtaaggcgc
     1441 ggctcatcgg gcggcgttcg ccacgatgtc ctgcacttcg agccaagcct cgaacacctg
     1501 ctggtgtgca cgactcaccc ggttgttgac accgcgcgcg gccgtgcggg ctcggtgggg
     1561 cggctctgtc gcccttgcca gcgtgagtag cgcgtacctc acctcgccca acaggtcgca
     1621 cacagccgat tcgtacgcca taaagccagg tgagcccacc agctccgtaa gttcgggcgc
     1681 tgtgtggctc gtacccgcgc attcaggcgg cagggggtct aacgggtcta aggcggcgtg
     1741 tacggccgcc acagcggctc tcagcggccc ggaaacgtcc tcgaaacgac gcatgtgttc
     1801 ctcctggttg gtacaggtgg ttgggggtgc tcggctgtcg ctggtgttcc accaccaggg
     1861 ctcgacggga gagcggggga gtgtgcagtt gtggggtggc ccctcagcga aatatctgac
     1921 ttggagctcg tgtcggacca tacaccggtg attaatcgtg gtctactacc aagcgtgagc
     1981 cacgtcgccg acgaatttga gcagctctgg ctgccgtact ggccgctggc aagcgacgat
     2041 ctgctcgagg ggatctaccg ccaaagccgc gcgtcggccc taggccgccg gtacatcgag
     2101 gcgaacccaa cagcgctggc aaacctgctg gtcgtggacg tagaccatcc agacgcagcg
     2161 ctccgagcgc tcagcgcccg ggggtcccat ccgctgccca acgcgatcgt gggcaatcgc
     2221 gccaacggcc acgcacacgc agtgtgggca ctcaacgccc ctgttccacg caccgaatac
     2281 gcgcggcgta agccgctcgc atacatggcg gcgtgcgccg aaggccttcg gcgcgccgtc
     2341 gacggcgacc gcagttactc aggcctcatg accaaaaacc ccggccacat cgcctgggaa
     2401 acggaatggc tccactcaga tctctacaca ctcagccaca tcgaggccga gctcggcgcg
     2461 aacatgccac cgccgcgctg gcgtcagcag accacgtaca aagcggctcc gacgccgcta
     2521 gggcggaatt gcgcactgtt cgattccgtc aggttgtggg cctatcgtcc cgccctcatg
     2581 cggatctacc tgccgacccg gaacgtggac ggactcggcc gcgcgatcta tgccgagtgc
     2641 cacgcgcgaa acgccgaatt cccgtgcaac gacgtgtgtc ccggaccgct accggacagc
     2701 gaggtccgcg ccatcgccaa cagcatttgg cgttggatca caaccaagtc gcgcatttgg
     2761 gcggacggga tcgtggtcta cgaggccaca ctcagtgcgc gccagtcggc catctcgcgg
     2821 aagggcgcag cagcgcgcac ggcggcgagc acagttgcgc ggcgcgcaaa gtccgcgtca
     2881 gccatggagg cattgctatg agcgacggct acagcgacgg ctacaaccgg cagccgactg
     2941 tccgcaaaaa gcggcgcgtg accgccgccg aaggcgctcg aatcaccgga ctatccgaac
     3001 gccacgtcgt ccggctcgtg gcgcaggaac gcagcgagtg gctcgccgag caggctgcac
     3061 gccgcgaacg catccgcgcc tatcacgacg acgagggcca ctcttggccg caaacggcca
     3121 aacatttcgg gctgcatctg gacaccgtta agcgactcgg ctatcgggcg aggaaagagc
     3181 gtgcggcaga acaggaagcg gctcaaaagg cccacaacga agccgacaat ccaccgctgt
     3241 tctaacgcaa ttggggagcg ggtgtcgcgg gggttccgtg gggggttccg ttgcaacggg
     3301 tcggacaggt aaaagtcctg gtagacgcta gttttctggt ttgggccatg cctgtctcgt
     3361 tgcgtgtttc gttgcgcccg ttttgaatac cagccagacg agacggggtt ctacgaatct
     3421 tggtcgatac caagccattt ccgctgaata tcggggagct caccgccaga atcggtggtt
     3481 gtggtgatgt acgtggcgaa ctccgttgta gtgcctgtgg tggcatccgt ggccactctc
     3541 gttgcacggt tcgttgtgcc gttacaggcc ccgttgacag ctcaccgaac gtagttaaaa
     3601 catgctggtc aaactaggtt taccaacgat acgagtcagc tcatctaggg ccagttctag
     3661 gcgttgttcg ttgcgcggtt cgttgcgcat gtttcgtgtg gttgctagat ggctccgcaa
     3721 ccacacgctt cgaggttgag tgcttccagc acgggcgcga tccagaagaa cttcgtcgtg
     3781 cgactgtcct cgttgatcct tgccgagctg ggatggaagc ccggccgacc caccctggag
     3841 gagatgatcg aggatgccag ggcctttcac gcccgccgct gctgagcgtc cgccgccggg
     3901 cccgcaccgc cgtcggccgg cccgctccgg gctcgcagca gcgggcttcg gcgcgggccc
     3961 ggggctcccg ggccgccggg cggggctccg cccggcggcc gccgggggcc gggggcggcg
     4021 ccgggcggcc cggggcgtca ggcgccgggg gcggtgtccg gcggccccca gaggaactgc
     4081 gccagttcct ccggatcggt gaagccggag agatccagcg gggtctcctc gaacacctcg
     4141 aagtcgtgca ggaaggtgaa ggcgagcagt tcgcgggcga agtcctcggt ccgcttccac
     4201 tgcgccccgt cgagcagcgc ggccaggatc tcgcggtcgc cccggaaggc gttgagatgc
     4261 agttgcacca ggctgtagcg ggagtctccc gcatagacgt cggtgaagtc gacgatcccg
     4321 gtgacctcgg tcgcggccag gtccacgaag atgttggtcc cgtgcaggtc gccgtggacg
     4381 aaccggggtt cgcggccggc cagcagcgtg tccacgtccg gcagccagtc ctccaggcgg
     4441 tccagcagcc ggggcgagag gtagccccac ccgcggtggt cctcgacggt cgccgcgcgg
     4501 cgttcccgca gcagttccgg gaagacctcg gaatgggggg tgagcacggt gttcccggtc
     4561 agcggcaccc tgtgcagccg gccgagcacc cggccgagtt cgcgggccag ggcgagcagc
     4621 gcgttccggt cggtcgtgcc gtccatcgcg gaccgccagg tggtgccggt catccggctc
     4681 atcaccaggt agggccacgg ccaggctccg gtgccgggcc gcagctcgcc gcggccgagg
     4741 aggcggggca ccggcaccgg ggcgtccgcc aggaccgcgt acgcctccga ctccgacgcg
     4801 aggctctccg gaccgcacca gtgctcgccg aacagcttga tcaccgggcc gggctcgccg
     4861 accagtacgg ggttggtgct ctcgccgggc acccgcagca ccggcggcac cggcagcccg
     4921 agctcctcca gggctcggcg ggccagcggc tcccagaatt cctggtcgtt ccgcaggctc
     4981 gcgtaggaat catccgaatc aatacggtcg agaagtaaca gggattcttg tgtcacagcg
     5041 gacctctatt cacagggtac gggccggctt aattccgcac ggccggtcgc gacacggcct
     5101 gtccgcaccg cggatcaggc gttgacgatg acgggctggt cggccacgtc ggggagtcag
     5161 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5221 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5281 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5341 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5401 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5461 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5521 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5581 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5641 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5701 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5761 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5821 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5881 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5941 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6001 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6061 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6121 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     6181 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctaga
//
