LOCUS       sequence-128280-        7207 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of 2GFP tag at Atg1 C terminus, use auxotrophic marker 
            URA3(Candida albicans).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7207)
  AUTHORS   Li D, Song JZ, Shan MH, Li SP, Liu W, Li H, Zhu J, Wang Y, Lin J, 
            Xie Z
  TITLE     A fluorescent tool set for yeast Atg proteins.
  JOURNAL   Autophagy. 2015;11(6):954-60. doi: 10.1080/15548627.2015.1040971.
  PUBMED    25998947
REFERENCE   2  (bases 1 to 7207)
  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: 
            "Autophagy."; date: "2015"; volume: "11(6):954-60. doi"; pages: " 
            10.1080/15548627.2015.1040971"
COMMENT     Sequence Label: LD201
FEATURES             Location/Qualifiers
     source          1..7207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             876..1589
                     /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             1626..2342
                     /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             1626..2339
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2363..2550
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2418..2442)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2625..2968
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3787..3984
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(4851..4870)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4852..4870)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4957..4974)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5128..5716)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5208..5227)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5887..6678,6679..6747))
                     /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     6510..6529
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6748..6852)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6920..6938
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6976..6998)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7098..7117
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(7198..7207,1..9)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(7198..7207,1..8)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 cactatagaa cgcggccgcc agctgaagct tcgtacgctg caggtcgagc agtatggata
       61 gtaatgctat tgcagaagaa caagactcag acgatgcaga agaagaggat gaaacgttga
      121 aaaagtacaa ggaagattgc ttatccacga aaacttttgg aaagggtaga actttatctg
      181 ccacatctca gttgagtgca actttcaata aactaccacg ttcggaaatg atccttctat
      241 gtaatgaggc cattgtctta tatatgaagg cattatccat tttatcaaaa tctatgcagg
      301 taacgtccaa ctggtggtat gaatctcaag aaaaatcatg ttctctaaga gttaacgtgt
      361 tggtacagtg gctaagggaa aaatttaatg aatgtttaga aaaagctgat ttcctgagat
      421 taaaaattaa cgacttgaga ttcaagcatg cttctgaggt agctgaaaat caaactttgg
      481 aagaaaaagg tagttcggaa gagccagtat atttagaaaa gctattatat gatcgtgcat
      541 tagaaatatc taagatggcc gcacatatgg aattaaaagg agaaaatttg tacaactgtg
      601 aattagctta tgcaacttca ctttggatgt tagaaacttc tctggacgat gatgatttta
      661 cgaatgccta tggtgactac ccttttaaaa ctaacataca tctaaaaagt aacgacgttg
      721 aagataaaga gaagtatcat agtgtgttgg atgaaaatga cagaataatc ataagaaagt
      781 atattgatag tattgcaaac aggttgaaaa tattgaggca gaagatgaac caccaaaatg
      841 gtcgacggat cggtgacggt gctggtttaa ttaacatgtc taaaggtgaa gaattattca
      901 ctggtgttgt cccaattttg gttgaattag atggtgatgt taatggtcac aaattttctg
      961 tctccggtga aggtgaaggt gatgctactt acggtaaatt gaccttaaaa tttatttgta
     1021 ctactggtaa attgccagtt ccatggccaa ccttagtcac tactttaact tatggtgttc
     1081 aatgtttttc tagataccca gatcatatga aacaacatga ctttttcaag tctgccatgc
     1141 cagaaggtta tgttcaagaa agaactattt ttttcaaaga tgacggtaac tacaagacca
     1201 gagctgaagt caagtttgaa ggtgatacct tagttaatag aatcgaatta aaaggtattg
     1261 attttaaaga agatggtaac attttaggtc acaaattgga atacaactat aactctcaca
     1321 atgtttacat catggctgac aaacaaaaga atggtatcaa agttaacttc aaaattagac
     1381 acaacattga agatggttct gttcaattag ctgaccatta tcaacaaaat actccaattg
     1441 gtgatggtcc agtcttgtta ccagacaacc attacttatc cactcaatct gccttatcca
     1501 aagatccaaa cgaaaagaga gaccacatgg tcttgttaga atttgttact gctgctggta
     1561 ttacccatgg tattgatgaa ttgtacaaaa ttccagttcc aacgcgtgat ctagagaagc
     1621 cattcatgtc taaaggtgaa gaattattca ctggtgttgt cccaattttg gttgaattag
     1681 atggtgatgt taatggtcac aaattttctg tctccggtga aggtgaaggt gatgctactt
     1741 acggtaaatt gaccttaaaa tttatttgta ctactggtaa attgccagtt ccatggccaa
     1801 ccttagtcac tactttaact tatggtgttc aatgtttttc tagataccca gatcatatga
     1861 aacaacatga ctttttcaag tctgccatgc cagaaggtta tgttcaagaa agaactattt
     1921 ttttcaaaga tgacggtaac tacaagacca gagctgaagt caagtttgaa ggtgatacct
     1981 tagttaatag aatcgaatta aaaggtattg attttaaaga agatggtaac attttaggtc
     2041 acaaattgga atacaactat aactctcaca atgtttacat catggctgac aaacaaaaga
     2101 atggtatcaa agttaacttc aaaattagac acaacattga agatggttct gttcaattag
     2161 ctgaccatta tcaacaaaat actccaattg gtgatggtcc agtcttgtta ccagacaacc
     2221 attacttatc cactcaatct gccttatcca aagatccaaa cgaaaagaga gaccacatgg
     2281 tcttgttaga atttgttact gctgctggta ttacccatgg tatggatgaa ttatataaat
     2341 aaggcgcgcc acttctaaat aagcgaattt cttatgattt atgattttta ttattaaata
     2401 agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa
     2461 attcttattc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg tatagtatga
     2521 ggtcgctctt attgaccaca cctctaccgg cagatccgct agggataaca gggtaatata
     2581 gatctgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg gaggcccaga
     2641 ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt
     2701 acatttagcc catacatccc catgtataat catttgcatc catacatttt gatggccgca
     2761 cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga aacgctcccc
     2821 tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata aaaggttagg
     2881 atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta ggatacagtt
     2941 ctcacatcac atccgaacat aaacaaccat gacagtcaac actaagacct atagtgagag
     3001 agcagaaact catgcctcac cagtagcaca gcgattattt cgattaatgg aactgaagaa
     3061 aaccaattta tgtgcatcaa ttgacgttga taccactaag gaattccttg aattaattga
     3121 taaattaggt ccttatgtat gcttaatcaa gactcatatt gatataatca atgatttttc
     3181 ctatgaatcc actattgaac cattattaga actttcacgt aaacatcaat ttatgatttt
     3241 tgaagataga aaatttgctg atattggtaa taccgtaaag aaacaatata ttggtggagt
     3301 ttataaaatt agtagttggg cagatattac caatgctcat ggtgtcactg ggaatggagt
     3361 ggttgaagga ttaaaacagg gagctaaaga aaccaccacc aaccaagagc caagagggtt
     3421 attgatgtta gctgaattat catcagtggg atcattagca tatggagaat attctcaaaa
     3481 aactgttgaa attgctaaat ccgataagga atttgttatt ggatttattg cccaacgtga
     3541 tatgggtggc caagaagaag gatttgattg gcttattatg acacctggag ttggattaga
     3601 tgataaaggt gatggattag gacaacaata tagaactgtt gatgaagttg ttagcactgg
     3661 aactgatatt atcattgttg gtagaggatt gtttggtaaa ggaagagatc cagatattga
     3721 aggtaaaagg tatagaaatg ctggttggaa tgcttatttg aaaaagactg gccaattata
     3781 atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt
     3841 tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc
     3901 gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt
     3961 atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca tccagtgtcg
     4021 aaaacgagct gtgatgcata atatggtttt cttattaagt acaaatgacc tgctatattt
     4081 tcaagtatct tcatgattat gttatgctac tctcgaaatg ctttacgagt tcgtcattaa
     4141 agtattcaaa ttttattttc cctttgtgtt ttgttttgtt tttatttcta cgtgattttt
     4201 ttcttgaaat ccagctcttt tatacctgga tgggttttat tttgatgttt aaatgttaat
     4261 taaggaaatt attgaattcc ttcggtttcg gtgtgttttt ttctttttaa gttttttttt
     4321 gttgcgttta gtacttgcct ttggctatac atatcctttt aatctaattt ttatttcttc
     4381 aaaatttgat tctttaacct tttgttctat atatatatat aaatgtatgt atgtatgatt
     4441 tgattgagga ggctttaaca agtcattaat atctttatta tttttacttg ttgagttcat
     4501 taaaattcat tttataattt gttgttacaa acagtatttt acaagtcata ataaacgttc
     4561 aaaaacaaac acaatattaa aagaattggt attaggatcc gacatgacaa ttaacttaga
     4621 tccttctatc aagtttctaa tcatatatta catctattaa aataatttac atatatcatg
     4681 ctaaagctta taaagagaca ttcttcttct cgtgcgctgg gcgtcgctag cctcaaatac
     4741 aagtgatgcg tctgcttcta ttctaaggtc atccttgctg ttagatagct cgaattcatc
     4801 gatgatatca gatccactag tggcctatgc ggccgcggat ctgccggtct ccctatagtg
     4861 agtcgtatta atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg
     4921 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4981 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5041 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5101 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5161 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5221 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5281 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     5341 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5401 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5461 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5521 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     5581 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5641 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5701 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5761 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5821 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5881 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5941 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6001 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6061 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6121 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6181 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6241 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6301 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6361 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6421 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6481 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6541 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6601 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6661 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6721 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6781 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6841 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     6901 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     6961 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     7021 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     7081 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatggacat
     7141 attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
     7201 taggtga
//
