LOCUS       sequence-121957-        7185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of GFP tag at Atg17 C terminus, use auxotrophic marker 
            URA3(Candida albicans).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7185)
  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 7185)
  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: LD117
FEATURES             Location/Qualifiers
     source          1..7185
                     /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"
     RBS             1413..1424
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1669..2385
                     /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             1669..2382
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2406..2593
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2461..2485)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2668..3011
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3830..4027
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(4951..4967)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4952..4968)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4997..5017)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4997..5015)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5036..5052)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5036..5052)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5049..5071)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5060..5076
                     /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(5084..5090,5091..5108,5109..5114))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5129..5150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5267..5284)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5438..6026)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5518..5537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6197..6988,6989..7057))
                     /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     6820..6839
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7058..7162)
                     /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 ccgcggaaag tacaaaggtt ggatggtata gtaaatgaat tgagcatatc agaaaaagat
      721 gataccgatc cctctaaact aggagattac atctcgagag acaacgttaa cttattgaat
      781 gacaaactga aagaagtgcc agtaattgag cgtcaaattg aaaacattaa gcttcaatac
      841 gaaaatatgg tcagaaaagt taacaaagaa ctgattgata ccaagttgac ggacgtaact
      901 caaaaattcc aaagtaaatt tggtatagat aacctgatgg aaacaaatgt tgcagagcag
      961 tttagcaggg aactaacgga ccttgaaaaa gatttagcag agataatgaa ttcattgaca
     1021 cagcactttg ataagacatt gctgttacaa gataaaaaaa ttgataacga tgaacgtgag
     1081 gagctgttta aggtggtaca aggcgacgac aaagaactat acaacatttt caaaactctg
     1141 catgaggtaa ttgatgacgt ggacaaaaca attcttaact tgggtcaatt tttgcaggca
     1201 aaaataaagg aaaagacaga actacacagc gaagtttctg aaataataaa cgatttcaat
     1261 agaaatttgg aatatctatt aatctttaaa gatatttcca atctgattga tagcttcaaa
     1321 aattcctgta cacaagatat tcaaacaact aaggaacttt gtgaatttta tgataatttt
     1381 gaagaaagct acggtaactt agttctagaa gcaaagagga gaaaggatgt ggcaaacaga
     1441 atgaaaacta tattgaaaga ttgtgaaaag cagttgcaga atttagatgc tcaggaccag
     1501 gaagaacgtc agaattttat agcggaaaat ggaacttatc ttcctgagac aatctggccc
     1561 ggtaaaattg acgatttttc ttccctgtac actttaaatt acaacgtgaa gaatcctgct
     1621 tcgtacgctg caggtcgacg gatcggtgac ggtgctggtt taattaacat gtctaaaggt
     1681 gaagaattat tcactggtgt tgtcccaatt ttggttgaat tagatggtga tgttaatggt
     1741 cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa attgacctta
     1801 aaatttattt gtactactgg taaattgcca gttccatggc caaccttagt cactacttta
     1861 acttatggtg ttcaatgttt ttctagatac ccagatcata tgaaacaaca tgactttttc
     1921 aagtctgcca tgccagaagg ttatgttcaa gaaagaacta tttttttcaa agatgacggt
     1981 aactacaaga ccagagctga agtcaagttt gaaggtgata ccttagttaa tagaatcgaa
     2041 ttaaaaggta ttgattttaa agaagatggt aacattttag gtcacaaatt ggaatacaac
     2101 tataactctc acaatgttta catcatggct gacaaacaaa agaatggtat caaagttaac
     2161 ttcaaaatta gacacaacat tgaagatggt tctgttcaat tagctgacca ttatcaacaa
     2221 aatactccaa ttggtgatgg tccagtcttg ttaccagaca accattactt atccactcaa
     2281 tctgccttat ccaaagatcc aaacgaaaag agagaccaca tggtcttgtt agaatttgtt
     2341 actgctgctg gtattaccca tggtatggat gaattgtaca aataaggcgc gccacttcta
     2401 aataagcgaa tttcttatga tttatgattt ttattattaa ataagttata aaaaaaataa
     2461 gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta ttcttgagta
     2521 actctttcct gtaggtcagg ttgctttctc aggtatagta tgaggtcgct cttattgacc
     2581 acacctctac cggcagatcc gctagggata acagggtaat atagatctgt ttagcttgcc
     2641 tcgtccccgc cgggtcaccc ggccagcgac atggaggccc agaataccct ccttgacagt
     2701 cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta gcccatacat
     2761 ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg aagcaaaaat
     2821 tacggctcct cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat
     2881 tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc actgaggttc
     2941 ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat cacatccgaa
     3001 cataaacaac catgacagtc aacactaaga cctatagtga gagagcagaa actcatgcct
     3061 caccagtagc acagcgatta tttcgattaa tggaactgaa gaaaaccaat ttatgtgcat
     3121 caattgacgt tgataccact aaggaattcc ttgaattaat tgataaatta ggtccttatg
     3181 tatgcttaat caagactcat attgatataa tcaatgattt ttcctatgaa tccactattg
     3241 aaccattatt agaactttca cgtaaacatc aatttatgat ttttgaagat agaaaatttg
     3301 ctgatattgg taataccgta aagaaacaat atattggtgg agtttataaa attagtagtt
     3361 gggcagatat taccaatgct catggtgtca ctgggaatgg agtggttgaa ggattaaaac
     3421 agggagctaa agaaaccacc accaaccaag agccaagagg gttattgatg ttagctgaat
     3481 tatcatcagt gggatcatta gcatatggag aatattctca aaaaactgtt gaaattgcta
     3541 aatccgataa ggaatttgtt attggattta ttgcccaacg tgatatgggt ggccaagaag
     3601 aaggatttga ttggcttatt atgacacctg gagttggatt agatgataaa ggtgatggat
     3661 taggacaaca atatagaact gttgatgaag ttgttagcac tggaactgat attatcattg
     3721 ttggtagagg attgtttggt aaaggaagag atccagatat tgaaggtaaa aggtatagaa
     3781 atgctggttg gaatgcttat ttgaaaaaga ctggccaatt ataatcagta ctgacaataa
     3841 aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct
     3901 attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga
     3961 tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc
     4021 tatactgctg tcgattcgat actaacgccg ccatccagtg tcgaaaacga gctcgaattc
     4081 atcgatgata tcagatccac tagtggccta tgctagaaaa aaggaaaaaa aggatacggt
     4141 acaaagattc aataattgta tcaatatttc gaaaaatact ttatcaaaaa agcaatactt
     4201 gaaggagaaa ggaaaaaatt cgctgtcttc ttcactttta cgacaaagaa aaatgtgccc
     4261 ttttcaacta aaagtcatga atgaataaat aaatgattac atatgcgtat attttatagt
     4321 gaacaacttc tgacatatta tcccgaatgt gagtaatcat gcttgtgctg tgaatctgtg
     4381 ttggtgtttg gctgtgaccc tgccaaaaag gtgctcgaag tgccggaaac aattccactt
     4441 ttatccaaac tgctcgtagt ggacgagtgg gataacgttc ttgttaaata ttttccaaaa
     4501 agtaaactgt ttgggccaaa tgagtcgcta tgttctaata caaattggct atgattggac
     4561 cctgaggtgt gatgatccct tccagaaaag cttgcctttt tattggggtt cttgtgcgaa
     4621 attattcgag cagtaatcat gttattcctg ctacgattgg cctcactgag agtacgacta
     4681 tatctttcta tctcctcagc gaaattttcc tcaaaaagac gcactagaag atcatgtgat
     4741 ggttttagat cctttgatgg tagtagttct gcatgcaacg ttagacagcg acctaatact
     4801 gcaacaagct catcaaaagc tagcgtcaat cttcctaaat catcagtact gaactgcttg
     4861 gatgtggagg gttctaaaaa ggccttgtac tgtgatatgc ctccattaac tgggatcccc
     4921 cgggctgcag gaattcgata tcaagcttat cgataccgtc gacctcgagg gggggcccgg
     4981 tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     5041 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5101 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5161 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5221 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5281 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5341 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5401 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5461 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5521 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5581 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5641 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5701 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5761 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5821 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     5881 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5941 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6001 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6061 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6121 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6181 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6241 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6301 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6361 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6421 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6481 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6541 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6601 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6661 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6721 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6781 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6841 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6901 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6961 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7021 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7081 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7141 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccac
//
