LOCUS       sequence-128283-        7624 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of 2GFP tag at Atg5 C terminus, use auxotrophic marker 
            URA3(Candida albicans).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7624)
  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 7624)
  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: LD205
FEATURES             Location/Qualifiers
     source          1..7624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1400..2113
                     /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             2150..2866
                     /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             2150..2863
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2887..3074
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2942..2966)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        3149..3492
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4311..4508
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(5436..5456)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5436..5454)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5475..5491)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5475..5491)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5488..5510)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5499..5515
                     /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)."
     promoter        complement(join(5523..5529,5530..5547,5548..5553))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5568..5589
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5706..5723)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5877..6465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5957..5976)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6636..7427,7428..7496))
                     /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     7259..7278
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7497..7601)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggcagt gagaagtatt tctggtttga gcataataag acgcctattc cgtggaatta
      721 cccagtaggt gttttgtttg actgcctagc cggaaaaagc gctactttta ccacctcttt
      781 tgagaaccag gtaaaggatg ttctcacttt tttgagaatt cacttggtta tgggcgattc
      841 gttgccacca actatcattc ccatcgcatc tagcaaaacg caagcggaaa agttttggtt
      901 tcaccaatgg aaacaagttt gcttcatact gaatggttcc tcaaaagcta ttatgtcttt
      961 atcggtcaac gaagctcgaa aattttgggg cagtgttatt acgagaaatt tccaagattt
     1021 cattgaaata tctaacaaga taagttcatc aagaccgcga catataccgc tcatcattca
     1081 aacctcaagg acatcaggaa cttttcgaat atcacaacca actatcagta tgactggagt
     1141 caatcctacg ttgaaggata ttgaaggcga tattctggac gtaaaggaag gaatcaatgg
     1201 taatgatgtc atggttattt gtcaaggaat tgaaattcct tggcatatgc tcctgtatga
     1261 tttgtattct aaattgcgaa gttttgatgg ctttttgtac ataactcttg ttcctataaa
     1321 aggcggcgat aaagcttcct ctgagctcgc ttcgtacgct gcaggtcgac ggatcggtga
     1381 cggtgctggt ttaattaaca tgtctaaagg tgaagaatta ttcactggtg ttgtcccaat
     1441 tttggttgaa ttagatggtg atgttaatgg tcacaaattt tctgtctccg gtgaaggtga
     1501 aggtgatgct acttacggta aattgacctt aaaatttatt tgtactactg gtaaattgcc
     1561 agttccatgg ccaaccttag tcactacttt aacttatggt gttcaatgtt tttctagata
     1621 cccagatcat atgaaacaac atgacttttt caagtctgcc atgccagaag gttatgttca
     1681 agaaagaact atttttttca aagatgacgg taactacaag accagagctg aagtcaagtt
     1741 tgaaggtgat accttagtta atagaatcga attaaaaggt attgatttta aagaagatgg
     1801 taacatttta ggtcacaaat tggaatacaa ctataactct cacaatgttt acatcatggc
     1861 tgacaaacaa aagaatggta tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg
     1921 ttctgttcaa ttagctgacc attatcaaca aaatactcca attggtgatg gtccagtctt
     1981 gttaccagac aaccattact tatccactca atctgcctta tccaaagatc caaacgaaaa
     2041 gagagaccac atggtcttgt tagaatttgt tactgctgct ggtattaccc atggtattga
     2101 tgaattgtac aaaattccag ttccaacgcg tgatctagag aagccattca tgtctaaagg
     2161 tgaagaatta ttcactggtg ttgtcccaat tttggttgaa ttagatggtg atgttaatgg
     2221 tcacaaattt tctgtctccg gtgaaggtga aggtgatgct acttacggta aattgacctt
     2281 aaaatttatt tgtactactg gtaaattgcc agttccatgg ccaaccttag tcactacttt
     2341 aacttatggt gttcaatgtt tttctagata cccagatcat atgaaacaac atgacttttt
     2401 caagtctgcc atgccagaag gttatgttca agaaagaact atttttttca aagatgacgg
     2461 taactacaag accagagctg aagtcaagtt tgaaggtgat accttagtta atagaatcga
     2521 attaaaaggt attgatttta aagaagatgg taacatttta ggtcacaaat tggaatacaa
     2581 ctataactct cacaatgttt acatcatggc tgacaaacaa aagaatggta tcaaagttaa
     2641 cttcaaaatt agacacaaca ttgaagatgg ttctgttcaa ttagctgacc attatcaaca
     2701 aaatactcca attggtgatg gtccagtctt gttaccagac aaccattact tatccactca
     2761 atctgcctta tccaaagatc caaacgaaaa gagagaccac atggtcttgt tagaatttgt
     2821 tactgctgct ggtattaccc atggtatgga tgaattatat aaataaggcg cgccacttct
     2881 aaataagcga atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata
     2941 agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt
     3001 aactctttcc tgtaggtcag gttgctttct caggtatagt atgaggtcgc tcttattgac
     3061 cacacctcta ccggcagatc cgctagggat aacagggtaa tatagatctg tttagcttgc
     3121 ctcgtccccg ccgggtcacc cggccagcga catggaggcc cagaataccc tccttgacag
     3181 tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca
     3241 tccccatgta taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa
     3301 ttacggctcc tcgctgcaga cctgcgagca gggaaacgct cccctcacag acgcgttgaa
     3361 ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt
     3421 cttctttcat atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga
     3481 acataaacaa ccatgacagt caacactaag acctatagtg agagagcaga aactcatgcc
     3541 tcaccagtag cacagcgatt atttcgatta atggaactga agaaaaccaa tttatgtgca
     3601 tcaattgacg ttgataccac taaggaattc cttgaattaa ttgataaatt aggtccttat
     3661 gtatgcttaa tcaagactca tattgatata atcaatgatt tttcctatga atccactatt
     3721 gaaccattat tagaactttc acgtaaacat caatttatga tttttgaaga tagaaaattt
     3781 gctgatattg gtaataccgt aaagaaacaa tatattggtg gagtttataa aattagtagt
     3841 tgggcagata ttaccaatgc tcatggtgtc actgggaatg gagtggttga aggattaaaa
     3901 cagggagcta aagaaaccac caccaaccaa gagccaagag ggttattgat gttagctgaa
     3961 ttatcatcag tgggatcatt agcatatgga gaatattctc aaaaaactgt tgaaattgct
     4021 aaatccgata aggaatttgt tattggattt attgcccaac gtgatatggg tggccaagaa
     4081 gaaggatttg attggcttat tatgacacct ggagttggat tagatgataa aggtgatgga
     4141 ttaggacaac aatatagaac tgttgatgaa gttgttagca ctggaactga tattatcatt
     4201 gttggtagag gattgtttgg taaaggaaga gatccagata ttgaaggtaa aaggtataga
     4261 aatgctggtt ggaatgctta tttgaaaaag actggccaat tataatcagt actgacaata
     4321 aaaagattct tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc
     4381 tattttaatc aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag
     4441 atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg
     4501 ctatactgct gtcgattcga tactaacgcc gccatccagt gtcgaaaacg agctcgaatt
     4561 catcgatgat atcagatcca ctagtggcct atgctaagag ctgttatata cgcatttaaa
     4621 agtgtcattc aaatatcgca gaaaataact tggacgttga aaatagggcc acatttatac
     4681 ggctgcaaga aaacttttac tctctttcta ccaagtttgt tactgcaatt aaataatctt
     4741 tatcatggga tagcgaaact aaaaactcat gagtggaaaa gagagaatcg tagtttttcg
     4801 agagctgttg atactttgga tacttttttc caaacatctt atcaaactct aactgaggta
     4861 cacctgtatc ggtttgtgtt ttatatagca ttccagcgta cagtacttgg gcaggaggta
     4921 gatcatgctt tgaaacacag caacacaacg ctttcaatga gcattccttc aaagcccaaa
     4981 ctcccgctat atattcatgc aatcgtggag ttaagcagtt ttcttcgatg agaagattac
     5041 tgaaatgaaa tctttccttt tcatgcatga acttccgtgt tattttattc aaggtagaac
     5101 ggccacatgg gtcgaatggg gaatatttct ctagtatatg tgcaaatctt ggcaagtata
     5161 caatgtcaac tcccactcct gctatcttac gtccaatatt cctcgatgca aaactcatca
     5221 ctggaggcat tcaaagctag cacttatgaa ataaacgttc attagaaagc aatactcctg
     5281 ataagaaaat agtgcttagc ttctattcag ccttcttaaa gaactagctt ttctatcaaa
     5341 agtcagatct cttatgaccg cttgaacctg tggaatgtat ataagcagtt ttgatgttaa
     5401 gactcgaggg ggggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
     5461 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5521 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5581 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5641 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5701 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5761 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5821 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5881 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5941 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6001 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6061 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6121 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6181 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6241 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6301 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6361 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6421 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6481 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6541 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6601 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6661 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6721 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6781 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6841 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6901 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6961 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7021 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7081 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7141 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7201 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7261 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7321 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7381 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7441 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7501 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7561 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7621 ccac
//
