LOCUS       sequence-62414-e        6759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6759)
  AUTHORS   Westermann B, Neupert W
  TITLE     Mitochondria-targeted green fluorescent proteins: convenient tools 
            for the study of organelle biogenesis in Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2000 Nov;16(15):1421-7.
  PUBMED    11054823
REFERENCE   2  (bases 1 to 6759)
  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: "Yeast."; 
            date: "2000-11"; volume: "16(15)"; pages: "1421-7"
COMMENT     Sequence Label: pYES-mtGFP
FEATURES             Location/Qualifiers
     source          1..6759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..444
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2..119
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     415..438
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     476..495
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        476..494
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             736..1449
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(766..794)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1389..1409
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1512..1759
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1528..1546)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1829..1846)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2000..2588)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2080..2099)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2759..3550,3551..3619))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLID*MEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(3715..4518)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4519..4739)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5338..6218
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(6287..6742)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     6636..6655
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 acggattaga agccgccgag cgggtgacag ccctccgaag gaagactctc ctccgtgcgt
       61 cctcgtcctc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga
      121 acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac
      181 ctggccccac aaaccttcaa atgaacgaat caaattaaca accataggat gataatgcga
      241 ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat
      301 taacagatat ataaatgcaa aaactgcatt aaccacttta actaatactt tcaacatttt
      361 cggtttgtat tacttcttat tcaaatgtaa taaaagtatc aacaaaaaat tgttaatata
      421 cctctatact ttaacgtcaa ggagaaaaaa ccccggatcg gactactagc agctgtaata
      481 cgactcacta tagggaatat taagcttatg gcctccactc gtgtcctcgc ctctcgcctg
      541 gcctcccgga tggctgcttc cgccaaggtt gcccgccctg ctgtccgcgt tgctcaggtc
      601 agcaagcgca ccatccagac tggctccccc ctccagaccc tcaagcgcac ccagatgacc
      661 tccatcgtca acgccaccac ccgccaggct ttccagaagc gcgcctactc ttccggatcc
      721 ccgggtacca gatctatgag taagggtgaa gaacttttca ctggagttgt cccaattctt
      781 gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt
      841 gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt
      901 ccatggccaa cacttgtcac tactttcact tatggtgttc aatgcttttc aagataccca
      961 gatcatatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa
     1021 agaactatat ttttcaaaga tgacgggaac tacaagacac gtgctgaagt caagtttgaa
     1081 ggtgataccc ttgttaatag aatcgagtta aaaggtattg attttaaaga agatggaaac
     1141 attcttggac acaaattgga atacaactat aactcacaca atgtatacat catggcagac
     1201 aaacaaaaga atggaatcaa agttaacttc aaaattagac acaacattga agatggaagc
     1261 gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta
     1321 ccagacaacc attacctgtc cacacaatct gccctttcga aagatcccaa cgaaaagaga
     1381 gaccacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg catggatgaa
     1441 ctatacaaat aagaattctg cagatatcca tcacactggc ggccgctcga gcatgcatct
     1501 agagggccgc atcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc
     1561 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     1621 tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta
     1681 cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg
     1741 ctcgaaggct ttaatttgcg gccctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1801 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1861 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1921 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaagcccagg aaccgtaaaa
     1981 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2041 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2101 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2161 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2221 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2281 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2341 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2401 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     2461 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2521 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2581 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2641 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2701 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2761 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2821 ttgcctgact ccccgtcgtg tagataacta cgatacggga gcgcttacca tctggcccca
     2881 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     2941 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccattcagt
     3001 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3061 ttgttggcat tgctacaggc atcgtggtgt cactctcgtc gtttggtatg gcttcattca
     3121 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3181 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3241 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3301 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3361 cttgcccggc gtcaatacgg gataatagtg tatcacatag cagaacttta aaagtgctca
     3421 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3481 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3541 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3601 ggaaatgttg aatactcata ctcttccttt ttcaatgggt aataactgat ataattaaat
     3661 tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag ttttttagtt
     3721 ttgctggccg catcttctca aatatgcttc ccagcctgct tttctgtaac gttcaccctc
     3781 taccttagca tcccttccct ttgcaaatag tcctcttcca acaataataa tgtcagatcc
     3841 tgtagagacc acatcatcca cggttctata ctgttgaccc aatgcgtctc ccttgtcatc
     3901 taaacccaca ccgggtgtca taatcaacca atcgtaacct tcatctcttc cacccatgtc
     3961 tctttgagca ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc
     4021 cttagtatat tctccagtag atagggagcc cttgcatgac aattctgcta acatcaaaag
     4081 gcctctaggt tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa caatacctgg
     4141 gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt atacacccgc
     4201 agagtactgc aatttgactg tattaccaat gtcagcaaat tttctgtctt cgaagagtaa
     4261 aaaattgtac ttggcggata atgcctttag cggcttaact gtgccctcca tggaaaaatc
     4321 agtcaagata tccacatgtg tttttagtaa acaaattttg ggacctaatg cttcaactaa
     4381 ctccagtaat tccttggtgg tacgaacatc caatgaagca cacaagtttg tttgcttttc
     4441 gtgcatgata ttaaatagct tggcagcaac aggactagga tgagtagcag cacgttcctt
     4501 atatgtagct ttcgacatga tttatcttcg tttcctgcag gtttttgttc tgtgcagttg
     4561 ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta tatataccaa
     4621 tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa
     4681 atttcaaaga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat tgaattgaaa
     4741 agctagctta tcgatgataa gctgtcaaag atgagaatta attccacgga ctatagacta
     4801 tactagatac tccgtctact gtacgataca cttccgctca ggtccttgtc ctttaacgag
     4861 gccttaccac tcttttgtta ctctattgat ccagctcagc aaaggcagtg tgatctaaga
     4921 ttctatcttc gcgatgtagt aaaactagct agaccgagaa agagactaga aatgcaaaag
     4981 gcacttctac aatggctgcc atcattatta tccgatgtga cgctgcagct tctcaatgat
     5041 attcgaatac gctttgagga gatacagcct aatatccgac aaactgtttt acagatttac
     5101 gatcgtactt gttacccatc attgaatttt gaacatccga acctgggagt tttccctgaa
     5161 acagatagta tatttgaacc tgtataataa tatatagtct agcgctttac ggaagacaat
     5221 gtatgtattt cggttcctgg agaaactatt gcatctattg cataggtaat cttgcacgtc
     5281 gcatccccgg ttcattttct gcgtttccat cttgcacttc aatagcatat ctttgttaac
     5341 gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa
     5401 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc gctattttac
     5461 caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgacg agagcgctaa
     5521 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc
     5581 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc
     5641 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg
     5701 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg
     5761 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta
     5821 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat
     5881 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag
     5941 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca
     6001 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa
     6061 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg
     6121 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga
     6181 tacttttgag caatgtttgt ggaagcggta ttcgcaatgg gaagctccac cccggttgat
     6241 aatcagaaaa gccccaaaaa caggaagatt gtataagcaa atatttaaat tgtaaacgtt
     6301 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taacgaatag
     6361 cccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     6421 gttccagttt ccaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     6481 aaaagggtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     6541 gggtcgaggt gccgtaaagc agtaaatcgg aagggtaaac ggatgccccc atttagagct
     6601 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggg
     6661 gctagggcgg tgggaagtgt aggggtcacg ctgggcgtaa ccaccacacc cgccgcgctt
     6721 aatggggcgc tacagggcgc gtggggatga tccactagt
//
