LOCUS       sequence-384003-        7729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Marker for yeast mitochondria. Integration of 2xGFP tag at ABF2 C 
            terminus. Uses auxotrophic marker URA3(Candida albicans). .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7729)
  AUTHORS   Li D, Yang SG, He CW, Zhang ZT, Liang Y, Li H, Zhu J, Su X, Gong Q, 
            Xie Z
  TITLE     Excess diacylglycerol at the endoplasmic reticulum disrupts 
            endomembrane homeostasis and autophagy.
  JOURNAL   BMC Biol. 2020 Aug 28;18(1):107. doi: 10.1186/s12915-020-00837-w.
  PUBMED    32859196
REFERENCE   2  (bases 1 to 7729)
  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: "BMC Biol.";
            date: "2020-08-28"; volume: "18(1):107. doi"; pages: " 
            10.1186/s12915-020-00837-w"
COMMENT     Sequence Label: ABF2-2EGFP-ter-URA3
FEATURES             Location/Qualifiers
     source          1..7729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1082..1795
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGIDELYK"
     CDS             1832..2548
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1832..2545
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2569..2756
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2624..2648)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    3039..3072
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      4027..4224
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    4278..4311
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(5373..5392)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5374..5392)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5479..5496)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5650..6238)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5730..5749)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6409..7200,7201..7269))
                     /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     7032..7051
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7270..7374)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7442..7460
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7498..7520)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7620..7639
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(7720..7729,1..9)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(7720..7729,1..8)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 cactatagaa cgcggccgcc agctgaagct taacgtggaa cagattcgct atcccgagtg
       61 caatacacaa gattatgttg atatatccaa accatctacc atatacgcta aagttgaaac
      121 ttttcaaatc cttcaccata ccacccacat tgatgaactg tcctaaactt aaagccatta
      181 ttgaaaacta gtatatttta gttgttattg ctcttctcct ggtgacctac aatcccactt
      241 cactagtagg ggaattctgg atctttactt cgctgatata gcaccttttt tctttttttt
      301 ttttcttttg ccttttttac gagccacaga ctttcctcct ttaccctact attcgtagcc
      361 gtcttttttt gttgtttttt ttttttgtga aacaacgtag caagaaaggg atcctcatcc
      421 aaactaggat aaacaagtaa acagattaac aaagaagcca atcaattaca acaacaaata
      481 aatgaacagt tacagcctat taactagatc tttccacgaa tcttctaagc cccttttcaa
      541 tttggctagc accttgttga aggcttccaa gagaacgcag cttagaaatg aattgataaa
      601 acagggtcct aaaaggccca catctgctta tttcttatat ttgcaagacc acagaagtca
      661 attcgttaag gaaaatccaa ccttacgtcc tgctgaaatc agcaagattg ccggtgaaaa
      721 gtggcaaaat ttagaggctg atataaagga aaaatacatt tctgaaagaa aaaagttgta
      781 ttctgaatac caaaaggcga agaaggagtt tgacgaaaaa cttcctccaa agaaaccagc
      841 aggacccttc attaagtatg ccaatgaagt tcgttcgcaa gtttttgcac aacatcctga
      901 caagtctcaa ttggatctaa tgaaaattat cggagataaa tggcaatcct tggatcaaag
      961 cattaaggac aaatacatac aagagtacaa aaaagctatc caagaatata atgctcgcta
     1021 ccctctcaac gcttcgtacg ctgcaggtcg acggatcggt gacggtgctg gtttaattaa
     1081 catgtctaaa ggtgaagaat tattcactgg tgttgtccca attttggttg aattagatgg
     1141 tgatgttaat ggtcacaaat tttctgtctc cggtgaaggt gaaggtgatg ctacttacgg
     1201 taaattgacc ttaaaattta tttgtactac tggtaaattg ccagttccat ggccaacctt
     1261 agtcactact ttaacttatg gtgttcaatg tttttctaga tacccagatc atatgaaaca
     1321 acatgacttt ttcaagtctg ccatgccaga aggttatgtt caagaaagaa ctattttttt
     1381 caaagatgac ggtaactaca agaccagagc tgaagtcaag tttgaaggtg ataccttagt
     1441 taatagaatc gaattaaaag gtattgattt taaagaagat ggtaacattt taggtcacaa
     1501 attggaatac aactataact ctcacaatgt ttacatcatg gctgacaaac aaaagaatgg
     1561 tatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggttctgttc aattagctga
     1621 ccattatcaa caaaatactc caattggtga tggtccagtc ttgttaccag acaaccatta
     1681 cttatccact caatctgcct tatccaaaga tccaaacgaa aagagagacc acatggtctt
     1741 gttagaattt gttactgctg ctggtattac ccatggtatt gatgaattgt acaaaattcc
     1801 agttccaacg cgtgatctag agaagccatt catgtctaaa ggtgaagaat tattcactgg
     1861 tgttgtccca attttggttg aattagatgg tgatgttaat ggtcacaaat tttctgtctc
     1921 cggtgaaggt gaaggtgatg ctacttacgg taaattgacc ttaaaattta tttgtactac
     1981 tggtaaattg ccagttccat ggccaacctt agtcactact ttaacttatg gtgttcaatg
     2041 tttttctaga tacccagatc atatgaaaca acatgacttt ttcaagtctg ccatgccaga
     2101 aggttatgtt caagaaagaa ctattttttt caaagatgac ggtaactaca agaccagagc
     2161 tgaagtcaag tttgaaggtg ataccttagt taatagaatc gaattaaaag gtattgattt
     2221 taaagaagat ggtaacattt taggtcacaa attggaatac aactataact ctcacaatgt
     2281 ttacatcatg gctgacaaac aaaagaatgg tatcaaagtt aacttcaaaa ttagacacaa
     2341 cattgaagat ggttctgttc aattagctga ccattatcaa caaaatactc caattggtga
     2401 tggtccagtc ttgttaccag acaaccatta cttatccact caatctgcct tatccaaaga
     2461 tccaaacgaa aagagagacc acatggtctt gttagaattt gttactgctg ctggtattac
     2521 ccatggtatg gatgaattat ataaataagg cgcgccactt ctaaataagc gaatttctta
     2581 tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa
     2641 agtgactctt aggttttaaa acgaaaattc ttattcttga gtaactcttt cctgtaggtc
     2701 aggttgcttt ctcaggtata gtatgaggtc gctcttattg accacacctc taccggcaga
     2761 tccgctaggg ataacagggt aatatagatc tgtttagctt gcctcgtccc cgccgggtca
     2821 cccggccagc gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc
     2881 aggggcatga tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt
     2941 tgcatccata cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgcg
     3001 gacctgcgag cagggaaacg ctcccctcac agacgcgtat aacttcgtat aatgtatgct
     3061 atacgaagtt attgaattgt ccccacgccg cgcccctgta gagaaatata aaaggttagg
     3121 atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta ggatacagtt
     3181 ctcacatcac atccgaacat aaacaaccat gacagtcaac actaagacct atagtgagag
     3241 agcagaaact catgcctcac cagtagcaca acgattattt cgattaatgg aactgaagaa
     3301 aaccaattta tgtgcatcaa ttgatgttga taccactaag gaattccttg aattaattga
     3361 taaattgggt ccttatgtat gcttaatcaa gactcatatt gatataatca atgatttttc
     3421 ctatgaatcc actattgaac cattattaga actttcacgt aaacatcaat ttatgatttt
     3481 tgaagataga aaatttgctg atattggtaa taccgtgaag aaacaatata ttggtggagt
     3541 ttataaaatt agtagttggg cagatattac taatgctcat ggtgtcactg ggaatggagt
     3601 agttgaagga ttaaaacagg gagctaaaga aaccaccacc aaccaagagc caagagggtt
     3661 attgatgtta gctgaattat catcagtggg atcattagca tatggagaat attctcaaaa
     3721 aactgttgaa attgctaaat ccgataagga atttgttatt ggatttattg cccaacgtga
     3781 tatgggtgga caagaagaag gatttgattg gcttattatg acacctggag ttggattaga
     3841 tgataaaggt gatggattag gacaacaata tagaactgtt gatgaagttg ttagcactgg
     3901 aactgatatt atcattgttg gtagaggatt gtttggtaaa ggaagagatc cagatattga
     3961 aggtaaaagg tatagagatg ctggttggaa tgcttatttg aaaaagactg gccaattata
     4021 atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt
     4081 tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc
     4141 gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt
     4201 atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca tccagtgtcg
     4261 aaaacgagct cgaattcata acttcgtata atgtatgcta tacgaagtta tatcgatgat
     4321 atcagatcca ctagtggcct atgcggccgc ggagaccgcg gtactctcac aatgtttttt
     4381 atgcctttat tctttccgtt cctatccaaa caaataaata aatttaactt aaccaagcaa
     4441 gtgtgtagtg tttttatatc ctatgtaagt cttcaaactt tttttttatc tctggtaacc
     4501 tttattcgca tacaacgcca atgaaacaaa cctgtaataa tttttcatag ttaacccatt
     4561 gatgatactt attaaaatat gtaatcaaat cgtcactatt acggtgcttt attaagatgt
     4621 ccgtaccacc tggatggaac ttcaaataac tagaaatgtc atagactttg ccgtttataa
     4681 cgcaccataa ctcgtcctca cctttacaat gcttctttac gatttttctg tttattctta
     4741 agggtggcat tattttataa aggggtatct tttggacaag caattggttc ttcagttgtg
     4801 aaggtgaaaa cttgaagtta tctttttgaa ataccggatc atcgagtaaa tcctggataa
     4861 gctgtagcga aacgaatttt gtataataat tcgctggatt ggaggttaga gagtgccagt
     4921 ccagagcact gtgcccagga tctagtctta ttttggttct tacaagaggc tttcttgacg
     4981 acttcgacat gggcatatgc agcggggatg atatggcagg atgagcaacg cttagtcgct
     5041 gaggagcagc gaatctgaca tttggtttcg agtttgaaac atctctgttc atgccgtcac
     5101 tactcatatc tctatgagtg taatctaagc acctgctaat atgttatatt ctgcttattc
     5161 gtccttttgc tacacataat taatgtaact ttaactctag ttctataaag gatatgattt
     5221 gtactaaatg taacattata cgaaaaaaca tattattcgt ttatataaac taatcaaaga
     5281 agtcgtcatc atcttcgtcg tcatcatttt tagcattctg cttgtcaaat gaattcagat
     5341 cttcgctgaa atctccgcgg atctgccggt ctccctatag tgagtcgtat taatttcgat
     5401 aagccaggtt aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5461 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5521 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5581 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5641 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5701 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5761 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5821 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5881 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5941 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6001 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6061 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6121 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6181 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6241 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6301 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6361 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6421 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6481 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6541 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6601 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6661 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6721 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6781 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6841 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6901 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6961 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7021 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7081 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7141 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7201 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7261 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7321 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7381 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     7441 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     7501 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     7561 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     7621 ggcatcagag cagattgtac tgagagtgca ccatatggac atattgtcgt tagaacgcgg
     7681 ctacaattaa tacataacct tatgtatcat acacatacga tttaggtga
//
