LOCUS       sequence-128241-        6874 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of GFP 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 6874)
  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 6874)
  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: LD105
FEATURES             Location/Qualifiers
     source          1..6874
                     /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..2116
                     /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      2137..2324
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2192..2216)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2399..2742
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3561..3758
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(4686..4706)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4686..4704)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4725..4741)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4725..4741)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4738..4760)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4749..4765
                     /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(4773..4779,4780..4797,4798..4803))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4818..4839
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4956..4973)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5127..5715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5207..5226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5886..6677,6678..6746))
                     /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     6509..6528
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6747..6851)
                     /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 aaataaggcg cgccacttct aaataagcga atttcttatg atttatgatt
     2161 tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag
     2221 gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct
     2281 caggtatagt atgaggtcgc tcttattgac cacacctcta ccggcagatc cgctagggat
     2341 aacagggtaa tatagatctg tttagcttgc ctcgtccccg ccgggtcacc cggccagcga
     2401 catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg
     2461 tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg catccataca
     2521 ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca
     2581 gggaaacgct cccctcacag acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa
     2641 tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt
     2701 gctaggatac agttctcaca tcacatccga acataaacaa ccatgacagt caacactaag
     2761 acctatagtg agagagcaga aactcatgcc tcaccagtag cacaacgatt atttcgatta
     2821 atggaactga agaaaaccaa tttatgtgca tcaattgatg ttgataccac taaggaattc
     2881 cttgaattaa ttgataaatt gggtccttat gtatgcttaa tcaagactca tattgatata
     2941 atcaatgatt tttcctatga atccactatt gaaccattat tagaactttc acgtaaacat
     3001 caatttatga tttttgaaga tagaaaattt gctgatattg gtaataccgt gaagaaacaa
     3061 tatattggtg gagtttataa aattagtagt tgggcagata ttactaatgc tcatggtgtc
     3121 actgggaatg gagtagttga aggattaaaa cagggagcta aagaaaccac caccaaccaa
     3181 gagccaagag ggttattgat gttagctgaa ttatcatcag tgggatcatt agcatatgga
     3241 gaatattctc aaaaaactgt tgaaattgct aaatccgata aggaatttgt tattggattt
     3301 attgcccaac gtgatatggg tggacaagaa gaaggatttg attggcttat tatgacacct
     3361 ggagttggat tagatgataa aggtgatgga ttaggacaac aatatagaac tgttgatgaa
     3421 gttgttagca ctggaactga tattatcatt gttggtagag gattgtttgg taaaggaaga
     3481 gatccagata ttgaaggtaa aaggtataga gatgctggtt ggaatgctta tttgaaaaag
     3541 actggccaat tataatcagt actgacaata aaaagattct tgttttcaag aacttgtcat
     3601 ttgtatagtt tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata
     3661 ttttttttcg cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat
     3721 catgcgtcaa tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc
     3781 gccatccagt gtcgaaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct
     3841 atgctaagag ctgttatata cgcatttaaa agtgtcattc aaatatcgca gaaaataact
     3901 tggacgttga aaatagggcc acatttatac ggctgcaaga aaacttttac tctctttcta
     3961 ccaagtttgt tactgcaatt aaataatctt tatcatggga tagcgaaact aaaaactcat
     4021 gagtggaaaa gagagaatcg tagtttttcg agagctgttg atactttgga tacttttttc
     4081 caaacatctt atcaaactct aactgaggta cacctgtatc ggtttgtgtt ttatatagca
     4141 ttccagcgta cagtacttgg gcaggaggta gatcatgctt tgaaacacag caacacaacg
     4201 ctttcaatga gcattccttc aaagcccaaa ctcccgctat atattcatgc aatcgtggag
     4261 ttaagcagtt ttcttcgatg agaagattac tgaaatgaaa tctttccttt tcatgcatga
     4321 acttccgtgt tattttattc aaggtagaac ggccacatgg gtcgaatggg gaatatttct
     4381 ctagtatatg tgcaaatctt ggcaagtata caatgtcaac tcccactcct gctatcttac
     4441 gtccaatatt cctcgatgca aaactcatca ctggaggcat tcaaagctag cacttatgaa
     4501 ataaacgttc attagaaagc aatactcctg ataagaaaat agtgcttagc ttctattcag
     4561 ccttcttaaa gaactagctt ttctatcaaa agtcagatct cttatgaccg cttgaacctg
     4621 tggaatgtat ataagcagtt ttgatgttaa gactcgaggg ggggcccggt acccagcttt
     4681 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     4741 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4801 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4861 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4921 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4981 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5041 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5101 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5161 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5221 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5281 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5341 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5401 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5461 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5521 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     5581 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5641 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5701 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5761 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5821 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5881 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5941 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6001 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6061 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6121 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6181 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6241 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6301 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6361 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6421 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6481 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6541 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6601 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6661 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6721 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6781 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6841 ggggttccgc gcacatttcc ccgaaaagtg ccac
//
