LOCUS       sequence-121952-        8434 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of GFP tag at Atg9 C terminus, use auxotrophic marker 
            URA3(Candida albicans).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8434)
  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 8434)
  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: LD109
FEATURES             Location/Qualifiers
     source          1..8434
                     /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             2209..2925
                     /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             2209..2922
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2946..3133
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3001..3025)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        3208..3551
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4370..4567
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(6246..6266)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6246..6264)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6285..6301)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6285..6301)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6298..6320)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6309..6325
                     /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(6333..6339,6340..6357,6358..6363))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6378..6399
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6516..6533)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6687..7275)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6767..6786)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7446..8237,8238..8306))
                     /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     8069..8088
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8307..8411)
                     /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 ccgcggttat gcgtcatggg ctttgtgttc aacgagtccg gatttattaa acaaagtatt
      721 ttaaaacctt ctcaaagaga attcacgaga gaagaattac aaaaaagatt catgctggca
      781 ggatttctta atattatatt ggcaccattt ctggtcacat actttgtttt gctttatttt
      841 ttcagatatt ttaatgagta taaaacttct cctggatcta ttggtgctcg tcaatacaca
      901 ccaattgctg aatggaaatt ccgtgagtac aatgaacttt accatatttt taagaaaaga
      961 ataagtttga gcacaacctt agctaacaag tatgttgatc agttcccgaa ggaaaagacc
     1021 aatttgtttc tgaaatttgt ttcctttatt tgtggatcat ttgtggccat tttagctttt
     1081 ctcactgtat tcgatccaga aaacttttta aactttgaaa tcacctcgga cagatctgtc
     1141 attttctaca ttactattct gggtgctata tggtctgtaa gtagaaatac aataacccag
     1201 gagtaccatg ttttcgaccc cgaagagacg ctcaaggagt tgtatgaata tacacactat
     1261 ctaccaaagg aatgggaagg aagatatcac aaagaggaaa tcaaactaga gttctgtaaa
     1321 ttgtacaact tgagaatagt aatacttttg agggaactca ctagcctgat gataacaccg
     1381 tttgtacttt ggttttcgtt gccctcgtca gccggcagga ttgtagattt cttcagagag
     1441 aattctgaat atgtggacgg attaggttac gtttgcaagt atgctatgtt taacatgaaa
     1501 aacatagacg gtgaagatac acacagcatg gatgaagaca gtttaacgaa aaagattgcc
     1561 gtgaacggaa gtcatacact caacagtaaa cgaaggagta aatttaccgc cgaagaccat
     1621 agcgataaag atttggcaaa taataaaatg ttacaatcat atgtatattt tatggatgat
     1681 tattccaaca gtgaaaactt aacagggaag taccaattgc ctgcaaaaaa aggctatccg
     1741 aataatgaag gcgattcatt tctcaacaat aagtattctt ggagaaagca atttcagcca
     1801 ggtcaaaagc cggagctatt tagaataggt aagcatgctc tcggccctgg ccataatatt
     1861 tcccctgcta tatattctac aaggaatccc ggcaagaact gggataacaa taataatggt
     1921 gatgacatta agaacgggac caataatgct acagctaaaa atgatgacaa taatggcaat
     1981 aatgatcatg aatacgtact aacagagtct ttccttgatt cgggcgcatt ccctaatcat
     2041 gatgtaatag accataataa aatgctaaac tcaaattaca atggtaatgg catactcaat
     2101 aagggtggtg tcttaggact tgttaaagag tattacaaga agtctgacgt cggaagagct
     2161 tcgtacgctg caggtcgacg gatcggtgac ggtgctggtt taattaacat gtctaaaggt
     2221 gaagaattat tcactggtgt tgtcccaatt ttggttgaat tagatggtga tgttaatggt
     2281 cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta cttacggtaa attgacctta
     2341 aaatttattt gtactactgg taaattgcca gttccatggc caaccttagt cactacttta
     2401 acttatggtg ttcaatgttt ttctagatac ccagatcata tgaaacaaca tgactttttc
     2461 aagtctgcca tgccagaagg ttatgttcaa gaaagaacta tttttttcaa agatgacggt
     2521 aactacaaga ccagagctga agtcaagttt gaaggtgata ccttagttaa tagaatcgaa
     2581 ttaaaaggta ttgattttaa agaagatggt aacattttag gtcacaaatt ggaatacaac
     2641 tataactctc acaatgttta catcatggct gacaaacaaa agaatggtat caaagttaac
     2701 ttcaaaatta gacacaacat tgaagatggt tctgttcaat tagctgacca ttatcaacaa
     2761 aatactccaa ttggtgatgg tccagtcttg ttaccagaca accattactt atccactcaa
     2821 tctgccttat ccaaagatcc aaacgaaaag agagaccaca tggtcttgtt agaatttgtt
     2881 actgctgctg gtattaccca tggtatggat gaattgtaca aataaggcgc gccacttcta
     2941 aataagcgaa tttcttatga tttatgattt ttattattaa ataagttata aaaaaaataa
     3001 gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta ttcttgagta
     3061 actctttcct gtaggtcagg ttgctttctc aggtatagta tgaggtcgct cttattgacc
     3121 acacctctac cggcagatcc gctagggata acagggtaat atagatctgt ttagcttgcc
     3181 tcgtccccgc cgggtcaccc ggccagcgac atggaggccc agaataccct ccttgacagt
     3241 cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta gcccatacat
     3301 ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg aagcaaaaat
     3361 tacggctcct cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat
     3421 tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc actgaggttc
     3481 ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat cacatccgaa
     3541 cataaacaac catgacagtc aacactaaga cctatagtga gagagcagaa actcatgcct
     3601 caccagtagc acagcgatta tttcgattaa tggaactgaa gaaaaccaat ttatgtgcat
     3661 caattgacgt tgataccact aaggaattcc ttgaattaat tgataaatta ggtccttatg
     3721 tatgcttaat caagactcat attgatataa tcaatgattt ttcctatgaa tccactattg
     3781 aaccattatt agaactttca cgtaaacatc aatttatgat ttttgaagat agaaaatttg
     3841 ctgatattgg taataccgta aagaaacaat atattggtgg agtttataaa attagtagtt
     3901 gggcagatat taccaatgct catggtgtca ctgggaatgg agtggttgaa ggattaaaac
     3961 agggagctaa agaaaccacc accaaccaag agccaagagg gttattgatg ttagctgaat
     4021 tatcatcagt gggatcatta gcatatggag aatattctca aaaaactgtt gaaattgcta
     4081 aatccgataa ggaatttgtt attggattta ttgcccaacg tgatatgggt ggccaagaag
     4141 aaggatttga ttggcttatt atgacacctg gagttggatt agatgataaa ggtgatggat
     4201 taggacaaca atatagaact gttgatgaag ttgttagcac tggaactgat attatcattg
     4261 ttggtagagg attgtttggt aaaggaagag atccagatat tgaaggtaaa aggtatagaa
     4321 atgctggttg gaatgcttat ttgaaaaaga ctggccaatt ataatcagta ctgacaataa
     4381 aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct
     4441 attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga
     4501 tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc
     4561 tatactgctg tcgattcgat actaacgccg ccatccagtg tcgaaaacga gctcgaattc
     4621 atcgatgata tcagatccac tagtggccta tgctaaggca gactgtgtcg tgtacatcat
     4681 ccaatataac tatatatata actgtttcct ttctttttcg tatttaccta aatgcatata
     4741 ttattcagtt cccactaatt ttgtactaca ataataagag aatcccttcc ctgttttaga
     4801 aatgagtagt tcttttcttt ttcataaata atacagttgt ctctgagaag cctaataaga
     4861 aaaaaagagg ttaataaatt attattattg gttatttctt tccgccacgt agttttcttt
     4921 ctctttcata cttgtttttt tccgcacctt cttcggtatt aatagtgtta acaatatgag
     4981 ccatctttgc atggtagtcg ctgctctcct tgttcttttc ttcaattctt tgtctagctt
     5041 caaatttggc gtctttacgg atttctttca tagtgaattt acgttccttc tttagttggg
     5101 ccttcatctt atttatctca cttcttgttc tatcaggatc gtacgatttt ttatcaggat
     5161 tgaagttttc ttcgtattta ggagcatgcg taggtatgga tacgggtttg tggttctgta
     5221 aagccaacgg aatatgttct gtgaatttcg ttaatttttc aactttgttc aaaatatttc
     5281 ttggcttttc aaaatcagaa tactttgagg tgtatgcact taataactgc tgaatcggta
     5341 aaataatttc attgaaagca ggtaagctct tccacacagt agagatagtt gcatctaaag
     5401 attccataac atttagtaaa acggagacac attgatcaac cggatgtgcc tcggtatcca
     5461 ttgttgacaa ggtatgtaat ggaataatag ttgatctctt ttttgtaaag tcaacgtcca
     5521 atggaagccc caactcgtaa gagtctagtc taatattttc aaaatctaat ggtttttctt
     5581 gattttcttt ctcaactata aaagtgagta atattttttg aaagaagtaa accacctccg
     5641 gtatgtaccg tttggaaatg cgttgatatt gtgaaacaat ccttaccaag acagcgccaa
     5701 aagctatcct ctttaaagaa ttaaacttga tttgttccag gaactggctc attaaaatca
     5761 atgctggagt tattactaaa tggtactggt ccgaggtgga gaaaagaata ccaataatgg
     5821 aaaaaaatac cagatcacca tttgatagag catcaaaatg gttcttttta tagcgtgctt
     5881 gcatttcatt gatataatct ctacactcct cggaaagctc tctattatat ttctctgaaa
     5941 gggattttag aattgagatc aatgcatttt gtgtacgttt aaaactttga acgtttttaa
     6001 ggtagttttg attgcttaga aaaatgatgt gtctcagtaa aacagcggtg aattttccta
     6061 acttttcctt attaccttca gccaattttg gttggtaagc tttgataata ttcttaacaa
     6121 tcttaggatg atcatctaag tctaattttt ttacttgatc taataacgca tcatgagttc
     6181 tagggcatga aatggaaatg ttctttttct ttctcgaggg ggggcccggt acccagcttt
     6241 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     6301 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6361 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6421 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6481 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6541 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6601 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6661 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6721 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6781 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6841 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6901 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6961 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7021 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7081 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     7141 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7201 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7261 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7321 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7381 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7441 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7501 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7561 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7621 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7681 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7741 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7801 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7861 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7921 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7981 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8041 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8101 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8161 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8221 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8281 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8341 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8401 ggggttccgc gcacatttcc ccgaaaagtg ccac
//
