LOCUS       sequence-128240-        6457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of GFP 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 6457)
  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 6457)
  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: LD101
FEATURES             Location/Qualifiers
     source          1..6457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             876..1592
                     /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"
     terminator      1613..1800
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1668..1692)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        1875..2218
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3037..3234
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(4101..4120)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4102..4120)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4207..4224)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4378..4966)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4458..4477)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5137..5928,5929..5997))
                     /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     5760..5779
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5998..6102)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6170..6188
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6226..6248)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6348..6367
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(6448..6457,1..9)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(6448..6457,1..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 ttgtacaaat aaggcgcgcc acttctaaat aagcgaattt
     1621 cttatgattt atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt
     1681 ttaaagtgac tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta
     1741 ggtcaggttg ctttctcagg tatagtatga ggtcgctctt attgaccaca cctctaccgg
     1801 cagatccgct agggataaca gggtaatata gatctgttta gcttgcctcg tccccgccgg
     1861 gtcacccggc cagcgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca
     1921 gctcaggggc atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat
     1981 catttgcatc catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc
     2041 tgcagacctg cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg
     2101 cgcccctgta gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac
     2161 ttccttttaa aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaccat
     2221 gacagtcaac actaagacct atagtgagag agcagaaact catgcctcac cagtagcaca
     2281 acgattattt cgattaatgg aactgaagaa aaccaattta tgtgcatcaa ttgatgttga
     2341 taccactaag gaattccttg aattaattga taaattgggt ccttatgtat gcttaatcaa
     2401 gactcatatt gatataatca atgatttttc ctatgaatcc actattgaac cattattaga
     2461 actttcacgt aaacatcaat ttatgatttt tgaagataga aaatttgctg atattggtaa
     2521 taccgtgaag aaacaatata ttggtggagt ttataaaatt agtagttggg cagatattac
     2581 taatgctcat ggtgtcactg ggaatggagt agttgaagga ttaaaacagg gagctaaaga
     2641 aaccaccacc aaccaagagc caagagggtt attgatgtta gctgaattat catcagtggg
     2701 atcattagca tatggagaat attctcaaaa aactgttgaa attgctaaat ccgataagga
     2761 atttgttatt ggatttattg cccaacgtga tatgggtgga caagaagaag gatttgattg
     2821 gcttattatg acacctggag ttggattaga tgataaaggt gatggattag gacaacaata
     2881 tagaactgtt gatgaagttg ttagcactgg aactgatatt atcattgttg gtagaggatt
     2941 gtttggtaaa ggaagagatc cagatattga aggtaaaagg tatagagatg ctggttggaa
     3001 tgcttatttg aaaaagactg gccaattata atcagtactg acaataaaaa gattcttgtt
     3061 ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat
     3121 gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt
     3181 gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg
     3241 attcgatact aacgccgcca tccagtgtcg aaaacgagct gtgatgcata atatggtttt
     3301 cttattaagt acaaatgacc tgctatattt tcaagtatct tcatgattat gttatgctac
     3361 tctcgaaatg ctttacgagt tcgtcattaa agtattcaaa ttttattttc cctttgtgtt
     3421 ttgttttgtt tttatttcta cgtgattttt ttcttgaaat ccagctcttt tatacctgga
     3481 tgggttttat tttgatgttt aaatgttaat taaggaaatt attgaattcc ttcggtttcg
     3541 gtgtgttttt ttctttttaa gttttttttt gttgcgttta gtacttgcct ttggctatac
     3601 atatcctttt aatctaattt ttatttcttc aaaatttgat tctttaacct tttgttctat
     3661 atatatatat aaatgtatgt atgtatgatt tgattgagga ggctttaaca agtcattaat
     3721 atctttatta tttttacttg ttgagttcat taaaattcat tttataattt gttgttacaa
     3781 acagtatttt acaagtcata ataaacgttc aaaaacaaac acaatattaa aagaattggt
     3841 attaggatcc gacatgacaa ttaacttaga tccttctatc aagtttctaa tcatatatta
     3901 catctattaa aataatttac atatatcatg ctaaagctta taaagagaca ttcttcttct
     3961 cgtgcgctgg gcgtcgctag cctcaaatac aagtgatgcg tctgcttcta ttctaaggtc
     4021 atccttgctg ttagatagct cgaattcatc gatgatatca gatccactag tggcctatgc
     4081 ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa gccaggttaa
     4141 cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4201 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4261 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4321 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4381 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4441 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4501 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4561 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4621 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4681 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4741 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4801 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4861 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4921 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4981 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5041 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5101 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5161 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5221 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5281 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5341 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5401 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5461 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5521 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5581 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5641 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5701 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5761 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5821 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5881 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5941 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6001 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6061 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6121 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     6181 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     6241 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     6301 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     6361 gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct acaattaata
     6421 cataacctta tgtatcatac acatacgatt taggtga
//
