LOCUS       sequence-351958-        4164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for genome integration in X2 site expressing sfpHluorin 
            (intracellular pH biosensor).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4164)
  AUTHORS   Torello Pianale L, Rugbjerg P, Olsson L
  TITLE     Real-Time Monitoring of the Yeast Intracellular State During 
            Bioprocesses With a Toolbox of Biosensors.
  JOURNAL   Front Microbiol. 2022 Jan 7;12:802169. doi: 
            10.3389/fmicb.2021.802169. eCollection 2021.
  PUBMED    35069506
REFERENCE   2  (bases 1 to 4164)
  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: "Front 
            Microbiol."; date: "2022-01-7"; volume: "12:802169. doi"; pages: " 
            10.3389/fmicb.2021.802169. eCollection 2021"
COMMENT     Sequence Label: LT1_34_sfpHluorin
FEATURES             Location/Qualifiers
     source          1..4164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        472..885
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             905..1618
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNEHLVYITADKQKNGTKA
                     IFQVHHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLHTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(935..963)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1558..1578
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1636..1652)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1637..1653)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1671..1689)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(join(2297..3088,3089..3157))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2920..2939
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3158..3262)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3365..3953
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3854..3873
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgaattc gcatctagat ggtagagcca caaacagccg gtacaagcaa cgatctccag
       61 gaccatctga atcatgcgcg gatgacacga actcacgacg gcgatcacag acattaaccc
      121 acagtacaga cactgcgaca acgtggcaat tcgtcgcaat accgtctcac tgaactggcc
      181 gataattgca gacgaacgcc ttgccaacag ggagttcttc agagacatgg aggctcaaaa
      241 cgaaattatt gacagcctag acatcaatag tcatacaaca gaaagcgacc acccaacttt
      301 ggctgataat agcgtataaa caatgcatac tttgtacgtt caaaatacaa tgcagtagat
      361 atatttatgc atattacata taatacatat cacataggaa gcaacaggcg cgttggactt
      421 ttaattttcg aggaccgcga atccttacat cacacccaat cccccacaag tgatccccca
      481 cacaccatag cttcaaaatg tttctactcc ttttttactc ttccagattt tctcggactc
      541 cgcgcatcgc cgtaccactt caaaacaccc aagcacagca tactaaattt cccctctttc
      601 ttcctctagg gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa aagacaccgc
      661 ctcgtttctt tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt tctttttctt
      721 gaaaattttt ttttttgatt tttttctctt tcgatgacct cccattgata tttaagttaa
      781 taaacggtca tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt
      841 ttacttcttg ctcattagaa agaaagcata gcaatctaat ctaagtttta attacaaaag
      901 atctatgagc aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga
      961 tggtgatgtt aatgggcaca aattttctgt ccgtggagag ggtgaaggtg atgctacaaa
     1021 cggaaaactc acccttaaat ttatttgcac tactggaaaa ctacctgttc cgtggccaac
     1081 acttgtcact actctgtctt atggtgttca atgcttttcc cgttatccgg atcacatgaa
     1141 acggcatgac tttttcaaga gtgccatgcc cgaaggttat gtacaggaac gcactatatc
     1201 tttcaaagat gacgggacct acaagacgcg tgctgaagtc aagtttgaag gtgataccct
     1261 tgttaatcgt atcgagttaa agggtattga ttttaaagaa gatggaaaca ttcttggaca
     1321 caaactcgag tacaacttta acgagcactt ggtatacatc acggcagaca aacaaaagaa
     1381 tggaaccaaa gctatcttcc aagttcacca caacgttgaa gatggttccg ttcaactagc
     1441 agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca
     1501 ttacctgcac acacaatctg tcctttcgaa agatcccaac gaaaagcgtg accacatggt
     1561 ccttcttgag tttgtaactg ctgctgggat tacacatggc atggatgagc tctacaaagg
     1621 atcctaactc gagatcgata ccgtcgacct cgagtcatgt aattagttat gtcacgctta
     1681 cattcacgcc ctcctcccac atccgctcta accgaaaagg aaggagttag acaacctgaa
     1741 gtctaggtcc ctatttattt tttttaatag ttatgttagt attaagaacg ttatttatat
     1801 ttcaaatttt tctttttttt ctgtacaaac gcgtgtacgc atgtaacatt atactgaaaa
     1861 ccttgcttga gaaggttttg ggacgctcgg ctgccaatga gacgacgggg tcatcacggc
     1921 tcatcatgcg ccaaacaaat gtgtgcaata cacgctcgga tgactgcatg atgaccgcac
     1981 tgactgggga cagcagatcc acctaagcct gtgagagaag cagacacccg acagatcaag
     2041 gcagttaact agtgcactgc agtacataac ccctaggtct tcctgagagt gacgaaggct
     2101 aaggtcactt cttcgtttcc tttattgggg tttccgtgta gccttcccct gaatagtgtg
     2161 ggacgtttta tgagaagccg taagaaatag ccgagcggcc gcgattatca aaaaggatct
     2221 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     2281 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     2341 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     2401 gcttaccatc tggccccagt gctgcaatga taccgcggga cccacgctca ccggctccag
     2461 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     2521 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     2581 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     2641 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     2701 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     2761 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     2821 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     2881 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     2941 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     3001 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     3061 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3121 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     3181 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     3241 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgtc atgaccaaaa
     3301 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3361 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3421 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3481 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3541 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3601 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3661 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3721 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3781 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3841 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3901 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3961 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4021 ctgcgttatc ccctgattct gtggataacc gtgcggccgc caatgtcaaa agatcctctc
     4081 ataccatatt aagtaaattg cctccatttc tttttcctcg ggcagagaaa ctcgcaggca
     4141 acttgctctc gaagtggtca cgtg
//
