LOCUS       sequence-238908-        6010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expressed pHiorine2 for pH quan. in gram-negative 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6010)
  AUTHORS   Arce-Rodriguez A, Volke DC, Bense S, Haussler S, Nikel PI
  TITLE     Non-invasive, ratiometric determination of intracellular pH in 
            Pseudomonas species using a novel genetically encoded indicator.
  JOURNAL   Microb Biotechnol. 2019 Jun 4. doi: 10.1111/1751-7915.13439.
  PUBMED    31162835
REFERENCE   2  (bases 1 to 6010)
  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: "Microb 
            Biotechnol. 2019 Jun 4. doi: 10.1111/1751-7915.13439."
COMMENT     Sequence Label: pS2513·PHP
FEATURES             Location/Qualifiers
     source          1..6010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..58
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     misc_feature    95..151
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      893..987
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1108..1127)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1119..1934
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            2101..2209
                     /note="incP origin of transfer"
     CDS             complement(2220..3071)
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(3058..3897)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             complement(4408..5379)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     rep_origin      5486..5880
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     terminator      complement(5918..6004)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaattaatt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
       61 ggtgaggaac taaaccccta ggccgcggcc gcgcgaattc gagctcggta cccggggatc
      121 ctctagagtc gacctgcagg catgcaagct taggaggaaa aacatatgag caaaggtgaa
      181 gaattgttca ccggtgtggt gccaattttg gtcgaactcg atggggatgt gaacggccat
      241 aagttctcgg tgagcggcga aggggaaggg gatgccacct atggcaaact cacgctcaaa
      301 tttatttgca ccaccggcaa gttgcctgtc ccctggccaa cgctggtcac cacgttgtcg
      361 tatggtgtcc aatgcttttc ccggtatcct gatcacatga aacaacatga cttttttaaa
      421 agcgcgatgc cggaagggta tgtgcaggag cggacgattt tttttaagga cgacgggaat
      481 tacaagacgc gggcagaggt gaagtttgaa ggtgacacgt tggtgaaccg tattgagttg
      541 aagggtatcg acttcaagga agatgggaac atcctcggtc acaaactgga atataattac
      601 aacgagcacc tcgtctacat tatggcggac aaacaaaaaa atggcacgaa agcgattttc
      661 caagtccacc ataacattga agatgggtcc gtccaactcg ccgaccatta ccaacagaat
      721 accccaatcg gcgatgggcc ggtgctgctc ccagacaacc actatctcca tacgcaatcg
      781 gcgctgtcca aggacccgaa tgaaaaacgt gatcatatgg tgctgctgga gttcgtcacc
      841 gcggctggta tcacccacgg tatggatgaa ctctacaagt aaactagtct tggactcctg
      901 ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc
      961 cgccgggcgt tttttattgg tgagaatcca ggggtcccca ataattacga tttaaatttg
     1021 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     1081 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt cagcgtgaaa
     1141 cgagctgtag ccgtccgcgt ctgaacagca acatggatgc ggatctgtat ggctataaat
     1201 gggcgcgtga taacgtgggt cagagcggcg cgaccattta tcgtctgtat ggcaaaccgg
     1261 atgcgccgga actgtttctg aaacatggca aaggcagcgt ggcgaacgat gtgaccgatg
     1321 aaatggtgcg tctgaactgg ctgaccgaat ttatgccgct gccgaccatt aaacatttta
     1381 ttcgcacccc ggatgatgcg tggctgctga ccaccgcgat tccgggcaaa accgcgtttc
     1441 aggtgctgga agaatatccg gatagcggcg aaaacattgt ggatgcgctg gccgtgtttc
     1501 tgcgtcgtct gcatagcatt ccggtgtgca actgcccgtt taacagcgat cgtgtgtttc
     1561 gtctggccca ggcgcagagc cgtatgaaca acggcctggt ggatgcgagc gattttgatg
     1621 atgaacgtaa cggctggccg gtggaacagg tgtggaaaga aatgcataaa ctgctgccgt
     1681 ttagcccgga tagcgtggtg acccacggcg attttagcct ggataacctg attttcgatg
     1741 aaggcaaact gattggctgc attgatgtgg gccgtgtggg cattgcggat cgttatcagg
     1801 atctggccat tctgtggaac tgcctgggcg aatttagccc gagcctgcaa aaacgtctgt
     1861 ttcagaaata tggcattgat aatccggata tgaacaaact gcaatttcat ctgatgctgg
     1921 atgaattttt ctaataatta attggaccgc ggtccgcgcg ttgtcctttt ccgctgcata
     1981 accctgcttc ggggtcatta tagcgatttt ttcggtatat ccatcctttt tcgcacgata
     2041 tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc
     2101 gggcaggata ggtgaagtag gcccacccgc gagcgggtgt tccttcttca ctgtccctta
     2161 ttcgcacctg gcggtgctca acgggaatcc tgctctgcga ggctggccgt aggccggcct
     2221 cagcctgccg ccttgggccg ggtgatgtcg tacttgcccg ccgcgaactc ggttaccgtc
     2281 cagcccagcg cgaccagctc cggcaacgcc tcgcgcaccc gctggcggcg cttgcgcatg
     2341 gtcgaaccac tggcctctga cggccagaca tagccgcaca aggtatctat ggaagccttg
     2401 ccggttttgc cggggtcgat ccagccacac agccgctggt gcagcaggcg ggcggtttcg
     2461 ctgtccagcg cccgcacctc gtccatgctg atgcgcacat gctggccgcc acccatgacg
     2521 gcctgcgcga tcaaggggtt cagggccacg tacaggcgcc cgtccgcctc gtcgctggcg
     2581 tactccgaca gcagccgaaa cccctgccgc ttgcggccat tctgggcgat gatggatacc
     2641 ttccaaaggc gctcgatgca gtcctgtatg tgcttgagcg ccccaccact atcgacctct
     2701 gccccgattt cctttgccag cgcccgatag ctacctttga ccaccatggc atcagcggtg
     2761 acggcctccc acttgggttc caggaacagc cggagctgcc gtccgccttc ggtcttgggt
     2821 tccgggccaa gcactaggcc attaggccca gccatggcca ccagcccttg caggatgcgc
     2881 agatcatcag cgcccagcgg ctccgggccg ctgaactcga tccgcttgcc gtcgccgtag
     2941 tcatacgtca cgtccagctt gctgcgcttg cgctcgcccc gcttgagggc acggaacagg
     3001 ccgggggcca gacagtgcgc cgggtcgtgc cggacgtggc tgaggctgtg cttgttctta
     3061 ggcttcacca cggggcaccc ccttgctctt gcgctgcctc tccagcacgg cgggcttgag
     3121 caccccgccg tcatgccgcc tgaaccaccg atcagcgaac ggtgcgccat agttggcctt
     3181 gctcacaccg aagcggacga agaaccggcg ctggtcgtcg tccacacccc attcctcggc
     3241 ctcggcgctg gtcatgctcg acaggtagga ctgccagcgg atgttatcga ccagtaccga
     3301 gctgccccgg ctggcctgct gctggtcgcc tgcgcccatc atggccgcgc ccttgctggc
     3361 atggtgcagg aacacgatag agcacccggt atcggcggcg atggcctcca tgcgaccgat
     3421 gacctgggcc atggggccgc tggcgttttc ttcctcgatg tggaaccggc gcagcgtgtc
     3481 cagcaccatc aggcggcggc cctcggcggc gcgcttgagg ccgtcgaacc actccggggc
     3541 catgatgttg ggcaggctgc cgatcagcgg ctggatcagc aggccgtcag ccacggcttg
     3601 ccgttcctcg gcgctgaggt gcgccccaag ggcgtgcagg cggtgatgaa tggcggtggg
     3661 cgggtcttcg gcgggcaggt agatcaccgg gccggtgggc agttcgccca cctccagcag
     3721 atccggcccg cctgcaatct gtgcggccag ttgcagggcc agcatggatt taccggcacc
     3781 accgggcgac accagcgccc cgaccgtacc ggccaccatg ttgggcaaaa cgtagtccag
     3841 cggtggcggc gctgctgcga acgcctccag aatattgata ggcttatggg tagccattga
     3901 ttgcctcctt tgcaggcagt tggtggttag gcgctggcgg ggtcactacc cccgccctgc
     3961 gccgctctga gttcttccag gcactcgcgc agcgcctcgt attcgtcgtc ggtcagccag
     4021 aacttgcgct gacgcatccc tttggccttc atgcgctcgg catatcgcgc ttggcgtaca
     4081 gcgtcagggc tggccagcag gtcgccggtc tgcttgtcct tttggtcttt catatcagtc
     4141 accgagaaac ttgccggggc cgaaaggctt gtcttcgcgg aacaaggaca aggtgcagcc
     4201 gtcaaggtta aggctggcca tatcagcgac tgaaaagcgg ccagcctcgg ccttgtttga
     4261 cgtataacca aagccaccgg gcaaccaata gcccttgtca cttttgatca ggtagaccga
     4321 ccctgaagcg cttttttcgt attccataaa acccccttct gtgcgtgagt actcatagta
     4381 taacaggcgt gagtaccaac gcaagcacta catgctgaaa tctggcccgc ccctgtccat
     4441 gcctcgctgg cggggtgccg gtgcccgtgc cagctcggcc cgcgcaagct ggacgctggg
     4501 cagacccatg accttgctga cggtgcgctc gatgtaatcc gcttcgtggc cgggcttgcg
     4561 ctctgccagc gctgggctgg cctcggccat ggccttgccg atttcctcgg cactgcggcc
     4621 ccggctggcc agcttctgcg cggcgataaa gtcgcacttg ctgaggtcat caccgaagcg
     4681 cttgaccagc ccggccatct cgctgcggta ctcgtccagc gccgtgcgcc ggtggcggct
     4741 aagctgccgc tcgggcagtt cgaggctggc cagcctgcgg gccttctcct gctgccgctg
     4801 ggcctgctcg atctgctggc cagcctgctg caccagcgcc gggccagcgg tggcggtctt
     4861 gcccttggat tcacgcagca gcacccacgg ctgataaccg gcgcgggtgg tgtgcttgtc
     4921 cttgcggttg gtgaagcccg ccaagcggcc atagtggcgg ctgtcggcgc tggccgggtc
     4981 ggcgtcgtac tcgctggcca gcgtccgggc aatctgcccc cgaagttcac cgcctgcggc
     5041 gtcggccacc ttgacccatg cctgatagtt cttcgggctg gtttccacta ccagggcagg
     5101 ctcccggccc tcggctttca tgtcatccag gtcaaactcg ctgaggtcgt ccaccagcac
     5161 cagaccatgc cgctcctgct cggcgggcct gatatacacg tcattgccct gggcattcat
     5221 ccgcttgagc catggcgtgt tctggagcac ttcggcggct gaccattccc ggttcatcat
     5281 ctggccggtg gtggcgtccc tgacgccgat atcgaagcgc tcacagccca tggccttgag
     5341 ctgtcggcct atggcctgca aagtcctgtc gttcttcatc gggccaccaa gcgattccca
     5401 cacattatac gagccggaag cataaagtgt agctagatcc gaaggatgag ccgggctgaa
     5461 tgatcgaccg agacaggccc tgcggggctg cacacgcgcc cccacccttc gggtaggggg
     5521 aaaggccgct aaagcggcta aaagcgctcc agcgtatttc tgcggggttt ggtgtggggt
     5581 ttagcgggct ttgcccgcct ttccccctgc cgcgcagcgg tggggcggtg tgtagcctag
     5641 cgcagcgaat agaccagcta tccggcctct ggccgggcat attgggcaag ggcagcagcg
     5701 ccccacaagg gcgctgataa ccgcgcctag tggattattc ttagataatc atggatggat
     5761 ttttccaaca ccccgccagc ccccgcccct gctgggtttg caggtttggg ggcgtgacag
     5821 ttattgcagg ggttcgtgac agttattgca ggggggcgtg acagttattg caggggttcg
     5881 tgacagttag ggcgcgccca gctgtctagg gcggcggatt tgtcctactc aggagagcgt
     5941 tcaccgacaa acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta
     6001 tttgatgcct
//
