LOCUS       sequence-128245-        7331 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of GFP tag at Atg14 C terminus, use auxotrophic marker 
            URA3(Candida albicans).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7331)
  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 7331)
  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: LD114
FEATURES             Location/Qualifiers
     source          1..7331
                     /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     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1147..1863
                     /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      1884..2071
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1939..1963)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2146..2489
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3308..3505
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(5143..5163)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5143..5161)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5182..5198)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5182..5198)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5195..5217)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5206..5222
                     /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(5230..5236,5237..5254,5255..5260))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5275..5296
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5413..5430)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5584..6172)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5664..5683)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6343..7134,7135..7203))
                     /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     6966..6985
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7204..7308)
                     /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 gccagtgaat tgtaatacga ctcactatag ggcgaattgg agctccaccg
      661 cggaacgatt ccatcaacat catgtggaag tatattagct tcttctcaga cattcttatg
      721 attaaacttc cctacacaaa taaaatctgt gaacaaccca tgtttgagtt ttccgatagc
      781 atacagacag tagtacaaag gttgatcaag cttatcataa atattttaca gatatgtaga
      841 catttgaaac tcgtaccttc aacacccatg gatatcccat ggctactgga ccagtacgat
      901 gtggatgggc tgttctataa tatggtgaaa cggaacaaga tgaagtgtag gtccgtctcg
      961 ctatattgga cttttgggat gttgtactcg atggttttgg ataacatgaa taatccacaa
     1021 agaggacatc ctgcaaggcg gaccgcacca cctccaacag tcacaggacc tcatgaccga
     1081 tggtacgtgg taggcgcttc gtacgctgca ggtcgacgga tcggtgacgg tgctggttta
     1141 attaacatgt ctaaaggtga agaattattc actggtgttg tcccaatttt ggttgaatta
     1201 gatggtgatg ttaatggtca caaattttct gtctccggtg aaggtgaagg tgatgctact
     1261 tacggtaaat tgaccttaaa atttatttgt actactggta aattgccagt tccatggcca
     1321 accttagtca ctactttaac ttatggtgtt caatgttttt ctagataccc agatcatatg
     1381 aaacaacatg actttttcaa gtctgccatg ccagaaggtt atgttcaaga aagaactatt
     1441 tttttcaaag atgacggtaa ctacaagacc agagctgaag tcaagtttga aggtgatacc
     1501 ttagttaata gaatcgaatt aaaaggtatt gattttaaag aagatggtaa cattttaggt
     1561 cacaaattgg aatacaacta taactctcac aatgtttaca tcatggctga caaacaaaag
     1621 aatggtatca aagttaactt caaaattaga cacaacattg aagatggttc tgttcaatta
     1681 gctgaccatt atcaacaaaa tactccaatt ggtgatggtc cagtcttgtt accagacaac
     1741 cattacttat ccactcaatc tgccttatcc aaagatccaa acgaaaagag agaccacatg
     1801 gtcttgttag aatttgttac tgctgctggt attacccatg gtattgatga attgtacaaa
     1861 taaggcgcgc cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat
     1921 aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa
     1981 aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagtatg
     2041 aggtcgctct tattgaccac acctctaccg gcagatccgc tagggataac agggtaatat
     2101 agatctgttt agcttgcctc gtccccgccg ggtcacccgg ccagcgacat ggaggcccag
     2161 aataccctcc ttgacagtct tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg
     2221 tacatttagc ccatacatcc ccatgtataa tcatttgcat ccatacattt tgatggccgc
     2281 acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct gcgagcaggg aaacgctccc
     2341 ctcacagacg cgttgaattg tccccacgcc gcgcccctgt agagaaatat aaaaggttag
     2401 gatttgccac tgaggttctt ctttcatata cttcctttta aaatcttgct aggatacagt
     2461 tctcacatca catccgaaca taaacaacca tgacagtcaa cactaagacc tatagtgaga
     2521 gagcagaaac tcatgcctca ccagtagcac aacgattatt tcgattaatg gaactgaaga
     2581 aaaccaattt atgtgcatca attgatgttg ataccactaa ggaattcctt gaattaattg
     2641 ataaattggg tccttatgta tgcttaatca agactcatat tgatataatc aatgattttt
     2701 cctatgaatc cactattgaa ccattattag aactttcacg taaacatcaa tttatgattt
     2761 ttgaagatag aaaatttgct gatattggta ataccgtgaa gaaacaatat attggtggag
     2821 tttataaaat tagtagttgg gcagatatta ctaatgctca tggtgtcact gggaatggag
     2881 tagttgaagg attaaaacag ggagctaaag aaaccaccac caaccaagag ccaagagggt
     2941 tattgatgtt agctgaatta tcatcagtgg gatcattagc atatggagaa tattctcaaa
     3001 aaactgttga aattgctaaa tccgataagg aatttgttat tggatttatt gcccaacgtg
     3061 atatgggtgg acaagaagaa ggatttgatt ggcttattat gacacctgga gttggattag
     3121 atgataaagg tgatggatta ggacaacaat atagaactgt tgatgaagtt gttagcactg
     3181 gaactgatat tatcattgtt ggtagaggat tgtttggtaa aggaagagat ccagatattg
     3241 aaggtaaaag gtatagagat gctggttgga atgcttattt gaaaaagact ggccaattat
     3301 aatcagtact gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt
     3361 ttatattgta gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct
     3421 cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg
     3481 tatgtgaatg ctggtcgcta tactgctgtc gattcgatac taacgccgcc atccagtgtc
     3541 gaaaacgagc tcgaattcat cgatgatatc agatccacta gtggcctatg ctagagtgcg
     3601 cacttttttt cctgccgtgt gtataaagtt gcatgtagtc agtcataaac aatagcagcc
     3661 agtgtaataa gcattcgggt aagaaccaac tcagtttttt agttgttttt tttttttttt
     3721 ttttcacccc gttgaaagaa aaggataaga gataaaggac taaatgattt taatacagat
     3781 agacaaccat tgttgacagg attgcgattg taagagtaga cagtacatca agcgaaaata
     3841 aatattgcag gaatggtttt gtctgataag gagttgtttg ccataaataa gaaagccgtc
     3901 gaacaaggtt tcaatgtgaa gcctagattg aactataata cggtcagtgg tgtgaacggt
     3961 ccattagtca ttttggaaaa ggtcaagttc ccacgttaca acgaaattgt taatttgaca
     4021 ttgccagatg gaaccgtgag acaaggtcaa gttttggaaa ttagaggaga tagagccatt
     4081 gtgcaagtgt ttgaaggtac atctggtatt gatgtcaaga agactaccgt ggaattcact
     4141 ggtgagagtt tgagaattcc tgtgtctgaa gacatgttgg gtagaatttt tgacggttct
     4201 ggtagaccca ttgacaacgg tcctaaagtt ttcgcagagg attacttgga cattaacggt
     4261 tctcctatca acccatatgc tcgtatttat ccagaagaaa tgatttctac tggtgtttct
     4321 gctattgaca caatgaactc cattgccaga ggtcaaaaga tcccaatttt ctccgcatca
     4381 ggtttaccac acaacgaaat tgcagcacaa atttgtagac aggctggttt ggtgagacct
     4441 accaaggatg ttcatgatgg tcatgaagaa aatttctcca tcgtttttgc tgccatgggt
     4501 gtcaacttgg aaaccgctag atttttcaaa caggatttcg aagaaaatgg gtctttggaa
     4561 agaacttcat tatttttgaa cttggctaat gaccctacca ttgaaagaat tatcactcca
     4621 agattggcct tgaccaccgc tgaatacctt gcttaccaaa cggaacgtca tgtgttgacc
     4681 atcttgaccg atatgtcatc gtatgctgat gctcttagag aagtttccgc tgctagagaa
     4741 gaagttccag gtagaagagg ttatcctggt tacatgtata cagatttgtc cacaatttat
     4801 gaaagagcag gtagagtaga gggtcgtaac gggtccatca ctcaaatacc tatcttgaca
     4861 atgcctaacg atgatattac gcatccaatt ccggatttga ccggttatat taccgagggt
     4921 caaatcttcg ttgaccgtca attacataac aagggtatct acccaccaat caacgtcttg
     4981 ccttcgttga gtagattgat gaaatctgcc atcggtgaag gtatgaccag aaaggaccac
     5041 ggtgacgttt ctaaccaatt gtatgccaag tacgccatcg gtaaggacgc tgctgctatg
     5101 aaggccgttc tcgagggggg gcccggtacc cagcttttgt tccctttagt gagggttaat
     5161 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5221 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5281 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5341 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5401 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5461 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5521 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5581 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5641 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5701 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5761 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5821 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5881 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5941 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6001 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     6061 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6121 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6181 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6241 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6301 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6361 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6421 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6481 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6541 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6601 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6661 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6721 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6781 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6841 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6901 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6961 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7021 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7081 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7141 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7201 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7261 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7321 aaaagtgcca c
//
