LOCUS       sequence-243934-        8032 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  super Ecliptic pHluorin with C terminal mRuby2 fusion tag in yeast 
            expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8032)
  AUTHORS   Dodd BJT, Kralj JM
  TITLE     Live Cell Imaging Reveals pH Oscillations in Saccharomyces 
            cerevisiae During Metabolic Transitions.
  JOURNAL   Sci Rep. 2017 Oct 24;7(1):13922. doi: 10.1038/s41598-017-14382-0.
  PUBMED    29066766
REFERENCE   2  (bases 1 to 8032)
  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: "Sci Rep."; 
            date: "2017-10-24"; volume: "7(1):13922. doi"; pages: " 
            10.1038/s41598-017-14382-0"
COMMENT     Sequence Label: pKL06
FEATURES             Location/Qualifiers
     source          1..8032
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(23..41)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    83..586
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        623..727
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(728..796,797..1588)
                     /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     complement(946..965)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1759..2347
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2248..2267
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2501..2518
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2671..2676,2677..2694,2695..2701)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2709..2725
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2714..2736
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2733..2749
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2733..2749
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2768..2788
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2770..2788
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2806..3204
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      4649..4896
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4665..4683)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4914..4933)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4915..4933)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4943..4960)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4943..4959)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4952..4974)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5104..5559
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5191..5210)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5401..5422
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5859..6263
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6276..7370
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(7863..7882)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7982..8004
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tgncttacnn tnagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
       61 ataataatgg tttcttagga cggatcgctt gcctgtaact tacacgcgcc tcgtatcttt
      121 taatgatgga ataatttggg aatttactct gtgtttattt atttttatgt tttgtatttg
      181 gattttagaa agtaaataaa gaaggtagaa gagttacgga atgaagaaaa aaaaataaac
      241 aaaggtttaa aaaatttcaa caaaaagcgt actttacata tatatttatt agacaagaaa
      301 agcagattaa atagatatac attcgattaa cgataagtaa aatgtaaaat cacaggattt
      361 tcgtgtgtgg tcttctacac agacaagatg aaacaattcg gcattaatac ctgagagcag
      421 gaagagcaag ataaaaggta gtatttgttg gcgatccccc tagagtcttt tacatcttcg
      481 gaaaacaaaa actatttttt ctttaatttc tttttttact ttctattttt aatttatata
      541 tttatattaa aaaatttaaa ttataattat ttttatagca cgtgatgaaa aggacccagg
      601 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
      661 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
      721 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
      781 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
      841 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
      901 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
      961 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1021 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1081 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1141 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1201 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1261 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1321 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1381 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1441 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     1501 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     1561 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     1621 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1681 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1741 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1801 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1861 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     1921 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1981 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2041 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2101 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2161 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2221 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2281 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2341 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2401 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2461 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2521 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     2581 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     2641 gagttacctc actcattagg caccccaggc tttacacttt atgcttccgg ctcctatgtt
     2701 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     2761 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctcatag cttcaaaatg
     2821 tttctactcc ttttttactc ttccagattt tctcggactc cgcgcatcgc cgtaccactt
     2881 caaaacaccc aagcacagca tactaaattt cccctctttc ttcctctagg gtgtcgttaa
     2941 ttacccgtac taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt tttcttcgtc
     3001 gaaaaaggca ataaaaattt ttatcacgtt tctttttctt gaaaattttt tttttgattt
     3061 ttttctcttt cgatgacctc ccattgatat ttaagttaat aaacggtctt caatttctca
     3121 agtttcagtt tcatttttct tgttctatta caactttttt tacttcttgc tcattagaaa
     3181 gaaagcatag caatctaatc taagttttct agagatgtct aagggcgagg aactgttcac
     3241 tggtgtggtt ccaatcctgg ttgagctgga cggcgatgta aacggtcaca agttcagcgt
     3301 tagcggcgaa ggtgagggcg acgctaccta cggtaaactg actctgaagt tcatctgcac
     3361 caccggtaaa ctgcctgttc cgtggccaac tctggtcacc accttcagct acggtgttca
     3421 gtgtctgacc cgctaccctg atcacatgaa acgccatgac ttcttcaaaa gcgcgatgcc
     3481 ggaaggctat gttcaagaac gtaccatctt tttcaaggat gacggcaatt ataaaactcg
     3541 tgccgaagta aagttcgaag gcgacaccct ggttaaccgt attgaactga aaggtattga
     3601 tttcaaagaa gatggtaaca ttctgggtca caaactggaa tataactaca acgaccacca
     3661 ggtttacatc atggcggata aacagaaaaa cggcatcaaa gctaacttca aaatccgtca
     3721 taacatcgaa gacggcggtg tgcagctggc tgaccactac cagcagaaca ccccgatcgg
     3781 tgatggtccg gttctgctgc cggataatca ctacctgttc accacgtcta ctctgtccaa
     3841 agacccgaac gaaaaacgtg atcacatggt cctgctggaa ttcgttactg cagcaggtat
     3901 cacccacggc atggatgaac tgtacaagat ggtttcaaag ggtgaagaac ttattaaaga
     3961 aaacatgaga atgaaggtag ttatggaagg aagcgttaat ggccatcaat tcaagtgtac
     4021 gggagaagga gagggtaggc cttacgaagg gacgcaaact atgaggatta aggttattga
     4081 gggtggtcca ttgccattcg cattcgacat cttagctaca tcttttatgt atggaagtcg
     4141 tacgttcata aagtatcctg ctgatattcc agacttcttt aaacagagct tccctgaggg
     4201 ctttacatgg gaaagagtga ccagatatga agatggtggt gttgttactg ttactcaaga
     4261 cacttcttta gaagacggtg aactagttta taatgtaaaa gttaggggtg ttaacttccc
     4321 gtctaacggg ccagtcatgc aaaagaagac aaaaggatgg gaacctaaca cggagatgat
     4381 gtatcccgct gacggtggtt tgagaggcta tacggatatt gcgttgaaag tcgacggagg
     4441 cggtcatttg cattgcaatt ttgttacaac ttacaggagt aagaagacag taggtaatat
     4501 caagatgccc ggcgttcacg cagttgacca tcgtttggaa cgtattgagg aatcagataa
     4561 tgagacatac gtggtacaga gggaggttgc cgtagctaag tacagtaact tggggggtgg
     4621 gatggacgaa ttatataaat aactcgagtc atgtaattag ttatgtcacg cttacattca
     4681 cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     4741 gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt
     4801 tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt
     4861 gagaaggttt tgggacgctc gaaggcttta atttgcggcc ggtacccaat tcgccctata
     4921 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     4981 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     5041 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     5101 gcgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     5161 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     5221 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     5281 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     5341 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     5401 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     5461 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     5521 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc ctgatgcggt attttctcct
     5581 tacgcatctg tgcggtattt cacaccgcat atcgacggtc gaggagaact tctagtatat
     5641 ccacatacct aatattattg ccttattaaa aatggaatcc caacaattac atcaaaatcc
     5701 acattctctt caaaatcaat tgtcctgtac ttccttgttc atgtgtgttc aaaaacgtta
     5761 tatttatagg ataattatac tctatttctc aacaagtaat tggttgtttg gccgagcggt
     5821 ctaaggcgcc tgattcaaga aatatcttga ccgcagttaa ctgtgggaat actcaggtat
     5881 cgtaagatgc aagagttcga atctcttagc aaccattatt tttttcctca acataacgag
     5941 aacacacagg ggcgctatcg cacagaatca aattcgatga ctggaaattt tttgttaatt
     6001 tcagaggtcg cctgacgcat ataccttttt caactgaaaa attgggagaa aaaggaaagg
     6061 tgagaggccg gaaccggctt ttcatataga atagagaagc gttcatgact aaatgcttgc
     6121 atcacaatac ttgaagttga caatattatt taaggaccta ttgttttttc caataggtgg
     6181 ttagcaatcg tcttactttc taacttttct taccttttac atttcagcaa tatatatata
     6241 tatttcaagg atataccatt ctaatgtctg cccctatgtc tgcccctaag aagatcgtcg
     6301 ttttgccagg tgaccacgtt ggtcaagaaa tcacagccga agccattaag gttcttaaag
     6361 ctatttctga tgttcgttcc aatgtcaagt tcgatttcga aaatcattta attggtggtg
     6421 ctgctatcga tgctacaggt gtcccacttc cagatgaggc gctggaagcc tccaagaagg
     6481 ttgatgccgt tttgttaggt gctgtgggtg gtcctaaatg gggtaccggt agtgttagac
     6541 ctgaacaagg tttactaaaa atccgtaaag aacttcaatt gtacgccaac ttaagaccat
     6601 gtaactttgc atccgactct cttttagact tatctccaat caagccacaa tttgctaaag
     6661 gtactgactt cgttgttgtc agagaattag tgggaggtat ttactttggt aagagaaagg
     6721 aagacgatgg tgatggtgtc gcttgggata gtgaacaata caccgttcca gaagtgcaaa
     6781 gaatcacaag aatggccgct ttcatggccc tacaacatga gccaccattg cctatttggt
     6841 ccttggataa agctaatgtt ttggcctctt caagattatg gagaaaaact gtggaggaaa
     6901 ccatcaagaa cgaattccct acattgaagg ttcaacatca attgattgat tctgccgcca
     6961 tgatcctagt taagaaccca acccacctaa atggtattat aatcaccagc aacatgtttg
     7021 gtgatatcat ctccgatgaa gcctccgtta tcccaggttc cttgggtttg ttgccatctg
     7081 cgtccttggc ctctttgcca gacaagaaca ccgcatttgg tttgtacgaa ccatgccacg
     7141 gttctgctcc agatttgcca aagaataagg ttgaccctat cgccactatc ttgtctgctg
     7201 caatgatgtt gaaattgtca ttgaacttgc ctgaagaagg taaggccatt gaagatgcag
     7261 ttaaaaaggt tttggatgca ggtatcagaa ctggtgattt aggtggttcc aacagtacca
     7321 ccgaagtcgg tgatgctgtc gccgaagaag ttaagaaaat ccttgcttaa aaagattctc
     7381 tttttttatg atatttgtac ataaacttta taaatgaaat tcataataga aacgacacga
     7441 aattacaaaa tggaatatgt tcatagggta gacgaaacta tatacgcaat ctacatacat
     7501 ttatcaagaa ggagaaaaag gaggatagta aaggaataca ggtaagcaaa ttgatactaa
     7561 tggctcaacg tgataaggaa aaagaattgc actttaacat taatattgac aaggaggagg
     7621 gcaccacaca aaaagttagg tgtaacagaa aatcatgaaa ctacgattcc taatttgata
     7681 ttggaggatt ttctctaaaa aaaaaaaaat acaacaaata aaaaacactc aatgacctga
     7741 ccatttgatg gagtttaagt caataccttc ttgaaccatt tcccataatg gtgaaagttc
     7801 cctcaagaat tttactctgt cagaaacggc cttacgacgt agtcgatatg gtgcactctc
     7861 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     7921 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     7981 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc ga
//
