LOCUS       sequence-210937-        8711 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 8711)
  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 8711)
  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: pYX142-mtGFP
FEATURES             Location/Qualifiers
     source          1..8711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             229..942
                     /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(259..287)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     882..902
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             967..993
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1121..1140)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1122..1140)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1147..1164)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1147..1163)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1156..1178)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1304..1759
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1391..1410)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1601..1622
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2060..2467
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2468..3562
                     /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
                     LIDSATMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(4056..4075)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4175..4197
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4295..4798
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4835..4939
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4940..5008,5009..5800)
                     /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"
     rep_origin      5971..6559
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6460..6479
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6713..6730
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6847..6868
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6883..6888,6889..6906,6907..6913)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6921..6937
                     /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     6926..6948
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6945..6961
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6945..6961
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6982..7000
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(7581..7685)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        8195..8703
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
ORIGIN
        1 atggcctcca ctcgtgtcct cgcctctcgc ctggcctccc ggatggctgc ttccgccaag
       61 gttgcccgcc ctgctgtccg cgttgctcag gtcagcaagc gcaccatcca gactggctcc
      121 cccctccaga ccctcaagcg cacccagatg acctccatcg tcaacgccac cacccgccag
      181 gctttccaga agcgcgccta ctcttccgga tccccgggta ccagatctat gagtaagggt
      241 gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg
      301 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt
      361 aaatttattt gcactactgg aaaactacct gttccatggc caacacttgt cactactttc
      421 acttatggtg ttcaatgctt ttcaagatac ccagatcata tgaaacggca tgactttttc
      481 aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg
      541 aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag
      601 ttaaaaggta ttgattttaa agaagatgga aacattcttg gacacaaatt ggaatacaac
      661 tataactcac acaatgtata catcatggca gacaaacaaa agaatggaat caaagttaac
      721 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa
      781 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
      841 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta
      901 acagctgctg ggattacaca tggcatggat gaactataca aataaaagct tacgcgtcga
      961 cccgggtatc cgtatgatgt gcctgactac gcatgatatc tcgagctcag ctagctaact
     1021 gaataaggaa caatgaacgt ttttcctttc tcttgttcct agtattaatg actgaccgat
     1081 acatcccttt ttttttttgt ctttgtctag ctccaattcg ccctatagtg agtcgtatta
     1141 caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     1201 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     1261 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc
     1321 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1381 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1441 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     1501 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1561 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1621 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     1681 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     1741 atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     1801 cacaccgcat atcgaccggt cgaggagaac ttctagtata tctacatacc taatattatt
     1861 gccttattaa aaatggaatc ccaacaatta catcaaaatc cacattctct tcaaaatcaa
     1921 ttgtcctgta cttccttgtt catgtgtgtt caaaaacgtt atatttatag gataattata
     1981 ctctatttct caacaagtaa ttggttgttt ggccgagcgg tctaaggcgc ctgattcaag
     2041 aaatatcttg accgcagtta actgtgggaa tactcaggta tcgtaagatg caagagttcg
     2101 aatctcttag caaccattat ttttttcctc aacataacga gaacacacag gggcgctatc
     2161 gcacagaatc aaattcgatg actggaaatt ttttgttaat ttcagaggtc gcctgacgca
     2221 tatacctttt tcaactgaaa aattgggaga aaaaggaaag gtgagagcgc cggaaccggc
     2281 ttttcatata gaatagagaa gcgttcatga ctaaatgctt gcatcacaat acttgaagtt
     2341 gacaatatta tttaaggacc tattgttttt tccaataggt ggttagcaat cgtcttactt
     2401 tctaactttt cttacctttt acatttcagc aatatatata tatatatttc aaggatatac
     2461 cattctaatg tctgccccta agaagatcgt cgttttgcca ggtgaccacg ttggtcaaga
     2521 aatcacagcc gaagccatta aggttcttaa agctatttct gatgttcgtt ccaatgtcaa
     2581 gttcgatttc gaaaatcatt taattggtgg tgctgctatc gatgctacag gtgttccact
     2641 tccagatgag gcgctggaag cctccaagaa ggctgatgcc gttttgttag gtgctgtggg
     2701 tggtcctaaa tggggtaccg gtagtgttag acctgaacaa ggtttactaa aaatccgtaa
     2761 agaacttcaa ttgtacgcca acttaagacc atgtaacttt gcatccgact ctcttttaga
     2821 cttatctcca atcaagccac aatttgctaa aggtactgac ttcgttgttg tcagagaatt
     2881 agtgggaggt atttactttg gtaagagaaa ggaagacgat ggtgatggtg tcgcttggga
     2941 tagtgaacaa tacaccgttc cagaagtgca aagaatcaca agaatggccg ctttcatggc
     3001 cctacaacat gagccaccat tgcctatttg gtccttggat aaagctaatg ttttggcctc
     3061 ttcaagatta tggagaaaaa ctgtggagga aaccatcaag aacgagttcc ctacattgaa
     3121 ggttcaacat caattgattg attctgccac catgatccta gttaagaacc caacccacct
     3181 aaatggtatt ataatcacca gcaacatgtt tggtgacatc atctccgatg aagcctccgt
     3241 tatcccaggt tccttgggtt tgttgccatc tgcgtccttg gcctctttgc cagacaagaa
     3301 caccgcattt ggtttgtacg aaccatgcca cggttctgct ccagatttgc caaagaataa
     3361 ggtcaaccct atcgccacta tcttgtctgc tgcaatgatg ttgaaattgt cattgaactt
     3421 gcctgaagaa ggtaaggcca ttgaagatgc agttaaaaag gttttggatg caggtatcag
     3481 aactggtgat ttaggtggtt ccaacagtac caccgaagtc ggtgatgctg tcgccgaaga
     3541 agttaagaaa atccttgctt aaaaagattc tcttttttta tgatatttgt acataaactt
     3601 tataaatgaa attcataata gaaacgacac gaaattacaa aatggaatat gttcataggg
     3661 tagacgaaac tatatacgca atctacatac atttatcaag aaggagaaaa aggaggatgt
     3721 aaaggaatac aggtaagcaa attgatacta atggctcaac gtgataagga aaaagaattg
     3781 cactttaaca ttaatattga caaggaggag ggcaccacac aaaaagttag gtgtaacaga
     3841 aaatcatgaa actatgattc ctaatttata tattggagga ttttctctaa aaaaaaaaaa
     3901 atacaacaaa taaaaaacac tcaatgacct gaccatttga tggagtttaa gtcaatacct
     3961 tcttgaacca tttcccataa tggtgaaagt tccctcaaga attttactct gtcagaaacg
     4021 gccttaacga cgtagtcgat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4081 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4141 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4201 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     4261 gttaatgtca tgataataat ggtttcttag gacggatcgc ttgcctgtaa cttacacgcg
     4321 cctcgtatct tttaatgatg gaataatttg ggaatttact ctgtgtttat ttatttttat
     4381 gttttgtatt tggattttag aaagtaaata aagaaggtag aagagttacg gaatgaagaa
     4441 aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc gtactttaca tatatattta
     4501 ttagacaaga aaagcagatt aaatagatat acattcgatt aacgataagt aaaatgtaaa
     4561 atcacaggat tttcgtgtgt ggtcttctac acagacaaga tgaaacaatt cggcattaat
     4621 acctgagagc aggaagagca agataaaagg tagtatttgt tggcgatccc cctagagtct
     4681 tttacatctt cggaaaacaa aaactatttt ttctttaatt tcttttttta ctttctattt
     4741 ttaatttata tatttatatt aaaaaattta aattataatt atttttatag cacgtgatga
     4801 aaaggaccca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4861 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4921 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4981 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5041 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5101 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5161 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5221 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5281 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5341 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5401 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5461 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5521 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5581 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5641 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5701 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5761 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5821 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5881 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5941 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6001 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6061 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6121 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6181 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6241 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6301 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6361 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6421 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6481 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6541 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6601 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6661 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     6721 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     6781 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     6841 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     6901 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     6961 ccatgattac gccaagctcg aaattaaccc tcactaaagg gaacaaaagc tggtaccggg
     7021 ccggccgtcg agcttgatgg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7081 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7141 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7201 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7261 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7321 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7381 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7441 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7501 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7561 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7621 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7681 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     7741 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt ggggatctac
     7801 gtatggtcat ttcttcttca gattccctca tggagaaagt gcggcagatg tatatgacag
     7861 agtcgccagt ttccaagaga ctttattcag gcacttccat gataggcaag agagaagacc
     7921 cagagatgtt gttgtcctag ttacacatgg tatttattcc agagtattcc tgatgaaatg
     7981 gtttagatgg acatacgaag agtttgaatc gtttaccaat gttcctaacg ggagcgtaat
     8041 ggtgatggaa ctggacgaat ccatcaatag atacgtcctg aggaccgtgc tacccaaatg
     8101 gactgattgt gagggagacc taactacata gtgtttaaag attacggata tttaacttac
     8161 ttagaataat gccatttttt tgagttataa taatcctacg ttagtgtgag cgggatttaa
     8221 actgtgagga ccttaataca ttcagacact tctgcggtat caccctactt attcccttcg
     8281 agattatatc taggaaccca tcaggttggt ggaagattac ccgttctaag acttttcagc
     8341 ttcctctatt gatgttacac ctggacaccc cttttctggc atccagtttt taatcttcag
     8401 tggcatgtga gattctccga aattaattaa agcaatcaca caattctctc ggataccacc
     8461 tcggttgaaa ctgacaggtg gtttgttacg catgctaatg caaaggagcc tatatacctt
     8521 tggctcggct gctgtaacag ggaatataaa gggcagcata atttaggagt ttagtgaact
     8581 tgcaacattt actattttcc cttcttacgt aaatattttt ctttttaatt ctaaatcaat
     8641 ctttttcaat tttttgtttg tattcttttc ttgcttaaat ctataactac aaaaaacaca
     8701 tacaggaatt c
//
