LOCUS       sequence-247762-        7340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  super Ecliptic pHluorin in yeast expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7340)
  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 7340)
  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: pKL05
FEATURES             Location/Qualifiers
     source          1..7340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..29)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    71..574
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        611..715
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(716..784,785..1576)
                     /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(934..953)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1747..2335
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2236..2255
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2489..2506
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2623..2644
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2659..2664,2665..2682,2683..2689)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2697..2713
                     /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     2702..2724
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2721..2737
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2721..2737
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2756..2776
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2758..2776
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2794..3192
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      3969..4216
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3985..4003)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4234..4253)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4235..4253)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4263..4280)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4263..4279)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4272..4294)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4420..4875
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4507..4526)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4717..4738
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5176..5583
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5584..6678
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(7172..7191)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7291..7313
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
       61 tcttaggacg gatcgcttgc ctgtaactta cacgcgcctc gtatctttta atgatggaat
      121 aatttgggaa tttactctgt gtttatttat ttttatgttt tgtatttgga ttttagaaag
      181 taaataaaga aggtagaaga gttacggaat gaagaaaaaa aaataaacaa aggtttaaaa
      241 aatttcaaca aaaagcgtac tttacatata tatttattag acaagaaaag cagattaaat
      301 agatatacat tcgattaacg ataagtaaaa tgtaaaatca caggattttc gtgtgtggtc
      361 ttctacacag acaagatgaa acaattcggc attaatacct gagagcagga agagcaagat
      421 aaaaggtagt atttgttggc gatcccccta gagtctttta catcttcgga aaacaaaaac
      481 tattttttct ttaatttctt tttttacttt ctatttttaa tttatatatt tatattaaaa
      541 aatttaaatt ataattattt ttatagcacg tgatgaaaag gacccaggtg gcacttttcg
      601 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
      661 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
      721 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
      781 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
      841 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
      901 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
      961 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     1021 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1081 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1141 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1201 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     1261 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     1321 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     1381 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     1441 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     1501 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     1561 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     1621 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     1681 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     1741 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     1801 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     1861 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     1921 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     1981 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2041 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2101 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2161 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2221 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2281 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2341 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     2401 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     2461 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     2521 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     2581 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     2641 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     2701 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
     2761 taaccctcac taaagggaac aaaagctgga gctcatagct tcaaaatgtt tctactcctt
     2821 ttttactctt ccagattttc tcggactccg cgcatcgccg taccacttca aaacacccaa
     2881 gcacagcata ctaaatttcc cctctttctt cctctagggt gtcgttaatt acccgtacta
     2941 aaggtttgga aaagaaaaaa gagaccgcct cgtttctttt tcttcgtcga aaaaggcaat
     3001 aaaaattttt atcacgtttc tttttcttga aaattttttt tttgattttt ttctctttcg
     3061 atgacctccc attgatattt aagttaataa acggtcttca atttctcaag tttcagtttc
     3121 atttttcttg ttctattaca acttttttta cttcttgctc attagaaaga aagcatagca
     3181 atctaatcta agttttctag agtaaaacac caagaactta gtttcgatgt ctaagggcga
     3241 ggaactgttc actggtgtgg ttccaatcct ggttgagctg gacggcgatg taaacggtca
     3301 caagttcagc gttagcggcg aaggtgaggg cgacgctacc tacggtaaac tgactctgaa
     3361 gttcatctgc accaccggta aactgcctgt tccgtggcca actctggtca ccaccttcag
     3421 ctacggtgtt cagtgtctga cccgctaccc tgatcacatg aaacgccatg acttcttcaa
     3481 aagcgcgatg ccggaaggct atgttcaaga acgtaccatc tttttcaagg atgacggcaa
     3541 ttataaaact cgtgccgaag taaagttcga aggcgacacc ctggttaacc gtattgaact
     3601 gaaaggtatt gatttcaaag aagatggtaa cattctgggt cacaaactgg aatataacta
     3661 caacgaccac caggtttaca tcatggcgga taaacagaaa aacggcatca aagctaactt
     3721 caaaatccgt cataacatcg aagacggcgg tgtgcagctg gctgaccact accagcagaa
     3781 caccccgatc ggtgatggtc cggttctgct gccggataat cactacctgt tcaccacgtc
     3841 tactctgtcc aaagacccga acgaaaaacg tgatcacatg gtcctgctgg aattcgttac
     3901 tgcagcaggt atcacccacg gcatggatga actgtacaag tgaagcttat cgataccgtc
     3961 gactcgagtc atgtaattag ttatgtcacg cttacattca cgccctcccc ccacatccgc
     4021 tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat
     4081 agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag
     4141 acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc
     4201 gaaggcttta atttgcggcc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc
     4261 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     4321 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     4381 cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat
     4441 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4501 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4561 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4621 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4681 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4741 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4801 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4861 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4921 ccgcatatcg accggtcgag gagaacttct agtatatcta catacctaat attattgcct
     4981 tattaaaaat ggaatcccaa caattacatc aaaatccaca ttctcttcaa aatcaattgt
     5041 cctgtacttc cttgttcatg tgtgttcaaa aacgttatat ttataggata attatactct
     5101 atttctcaac aagtaattgg ttgtttggcc gagcggtcta aggcgcctga ttcaagaaat
     5161 atcttgaccg cagttaactg tgggaatact caggtatcgt aagatgcaag agttcgaatc
     5221 tcttagcaac cattattttt ttcctcaaca taacgagaac acacaggggc gctatcgcac
     5281 agaatcaaat tcgatgactg gaaatttttt gttaatttca gaggtcgcct gacgcatata
     5341 cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga gagcgccgga accggctttt
     5401 catatagaat agagaagcgt tcatgactaa atgcttgcat cacaatactt gaagttgaca
     5461 atattattta aggacctatt gttttttcca ataggtggtt agcaatcgtc ttactttcta
     5521 acttttctta ccttttacat ttcagcaata tatatatata tatttcaagg atataccatt
     5581 ctaatgtctg cccctaagaa gatcgtcgtt ttgccaggtg accacgttgg tcaagaaatc
     5641 acagccgaag ccattaaggt tcttaaagct atttctgatg ttcgttccaa tgtcaagttc
     5701 gatttcgaaa atcatttaat tggtggtgct gctatcgatg ctacaggtgt tccacttcca
     5761 gatgaggcgc tggaagcctc caagaaggct gatgccgttt tgttaggtgc tgtgggtggt
     5821 cctaaatggg gtaccggtag tgttagacct gaacaaggtt tactaaaaat ccgtaaagaa
     5881 cttcaattgt acgccaactt aagaccatgt aactttgcat ccgactctct tttagactta
     5941 tctccaatca agccacaatt tgctaaaggt actgacttcg ttgttgtcag agaattagtg
     6001 ggaggtattt actttggtaa gagaaaggaa gacgatggtg atggtgtcgc ttgggatagt
     6061 gaacaataca ccgttccaga agtgcaaaga atcacaagaa tggccgcttt catggcccta
     6121 caacatgagc caccattgcc tatttggtcc ttggataaag ctaatgtttt ggcctcttca
     6181 agattatgga gaaaaactgt ggaggaaacc atcaagaacg aattccctac attgaaggtt
     6241 caacatcaat tgattgattc tgccgccatg atcctagtta agaacccaac ccacctaaat
     6301 ggtattataa tcaccagcaa catgtttggt gatatcatct ccgatgaagc ctccgttatc
     6361 ccaggttcct tgggtttgtt gccatctgcg tccttggcct ctttgccaga caagaacacc
     6421 gcatttggtt tgtacgaacc atgccacggt tctgctccag atttgccaaa gaataaggtc
     6481 aaccctatcg ccactatctt gtctgctgca atgatgttga aattgtcatt gaacttgcct
     6541 gaagaaggta aggccattga agatgcagtt aaaaaggttt tggatgcagg tatcagaact
     6601 ggtgatttag gtggttccaa cagtaccacc gaagtcggtg atgctgtcgc cgaagaagtt
     6661 aagaaaatcc ttgcttaaaa agattctctt tttttatgat atttgtacat aaactttata
     6721 aatgaaattc ataatagaaa cgacacgaaa ttacaaaatg gaatatgttc atagggtaga
     6781 cgaaactata tacgcaatct acatacattt atcaagaagg agaaaaagga ggatgtaaag
     6841 gaatacaggt aagcaaattg atactaatgg ctcaacgtga taaggaaaaa gaattgcact
     6901 ttaacattaa tattgacaag gaggagggca ccacacaaaa agttaggtgt aacagaaaat
     6961 catgaaacta tgattcctaa tttatatatt ggaggatttt ctctaaaaaa aaaaaaatac
     7021 aacaaataaa aaacactcaa tgacctgacc atttgatgga gtttaagtca ataccttctt
     7081 gaaccatttc ccataatggt gaaagttccc tcaagaattt tactctgtca gaaacggcct
     7141 taacgacgta gtcgatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     7201 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     7261 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     7321 cgtcatcacc gaaacgcgcg
//
