LOCUS       sequence-121955-        7334 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 7334)
  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 7334)
  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..7334
                     /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             1150..1866
                     /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             1150..1863
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1887..2074
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1942..1966)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2149..2492
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3311..3508
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(5146..5166)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5146..5164)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5185..5201)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5185..5201)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5198..5220)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5209..5225
                     /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(5233..5239,5240..5257,5258..5263))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5278..5299
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5416..5433)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5587..6175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5667..5686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6346..7137,7138..7206))
                     /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     6969..6988
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7207..7311)
                     /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 ccgcggaacg attccatcaa catcatgtgg aagtatatta gcttcttctc agacattctt
      721 atgattaaac ttccctacac aaataaaatc tgtgaacaac ccatgtttga gttttccgat
      781 agcatacaga cagtagtaca aaggttgatc aagcttatca taaatatttt acagatatgt
      841 agacatttga aactcgtacc ttcaacaccc atggatatcc catggctact ggaccagtac
      901 gatgtggatg ggctgttcta taatatggtg aaacggaaca agatgaagtg taggtccgtc
      961 tcgctatatt ggacttttgg gatgttgtac tcgatggttt tggataacat gaataatcca
     1021 caaagaggac atcctgcaag gcggaccgca ccacctccaa cagtcacagg acctcatgac
     1081 cgatggtacg tggtaggcgc ttcgtacgct gcaggtcgac ggatcggtga cggtgctggt
     1141 ttaattaaca tgtctaaagg tgaagaatta ttcactggtg ttgtcccaat tttggttgaa
     1201 ttagatggtg atgttaatgg tcacaaattt tctgtctccg gtgaaggtga aggtgatgct
     1261 acttacggta aattgacctt aaaatttatt tgtactactg gtaaattgcc agttccatgg
     1321 ccaaccttag tcactacttt aacttatggt gttcaatgtt tttctagata cccagatcat
     1381 atgaaacaac atgacttttt caagtctgcc atgccagaag gttatgttca agaaagaact
     1441 atttttttca aagatgacgg taactacaag accagagctg aagtcaagtt tgaaggtgat
     1501 accttagtta atagaatcga attaaaaggt attgatttta aagaagatgg taacatttta
     1561 ggtcacaaat tggaatacaa ctataactct cacaatgttt acatcatggc tgacaaacaa
     1621 aagaatggta tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg ttctgttcaa
     1681 ttagctgacc attatcaaca aaatactcca attggtgatg gtccagtctt gttaccagac
     1741 aaccattact tatccactca atctgcctta tccaaagatc caaacgaaaa gagagaccac
     1801 atggtcttgt tagaatttgt tactgctgct ggtattaccc atggtatgga tgaattgtac
     1861 aaataaggcg cgccacttct aaataagcga atttcttatg atttatgatt tttattatta
     1921 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
     1981 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt
     2041 atgaggtcgc tcttattgac cacacctcta ccggcagatc cgctagggat aacagggtaa
     2101 tatagatctg tttagcttgc ctcgtccccg ccgggtcacc cggccagcga catggaggcc
     2161 cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc
     2221 ccgtacattt agcccataca tccccatgta taatcatttg catccataca ttttgatggc
     2281 cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca gggaaacgct
     2341 cccctcacag acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt
     2401 taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt gctaggatac
     2461 agttctcaca tcacatccga acataaacaa ccatgacagt caacactaag acctatagtg
     2521 agagagcaga aactcatgcc tcaccagtag cacagcgatt atttcgatta atggaactga
     2581 agaaaaccaa tttatgtgca tcaattgacg ttgataccac taaggaattc cttgaattaa
     2641 ttgataaatt aggtccttat gtatgcttaa tcaagactca tattgatata atcaatgatt
     2701 tttcctatga atccactatt gaaccattat tagaactttc acgtaaacat caatttatga
     2761 tttttgaaga tagaaaattt gctgatattg gtaataccgt aaagaaacaa tatattggtg
     2821 gagtttataa aattagtagt tgggcagata ttaccaatgc tcatggtgtc actgggaatg
     2881 gagtggttga aggattaaaa cagggagcta aagaaaccac caccaaccaa gagccaagag
     2941 ggttattgat gttagctgaa ttatcatcag tgggatcatt agcatatgga gaatattctc
     3001 aaaaaactgt tgaaattgct aaatccgata aggaatttgt tattggattt attgcccaac
     3061 gtgatatggg tggccaagaa gaaggatttg attggcttat tatgacacct ggagttggat
     3121 tagatgataa aggtgatgga ttaggacaac aatatagaac tgttgatgaa gttgttagca
     3181 ctggaactga tattatcatt gttggtagag gattgtttgg taaaggaaga gatccagata
     3241 ttgaaggtaa aaggtataga aatgctggtt ggaatgctta tttgaaaaag actggccaat
     3301 tataatcagt actgacaata aaaagattct tgttttcaag aacttgtcat ttgtatagtt
     3361 tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata ttttttttcg
     3421 cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa
     3481 tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc gccatccagt
     3541 gtcgaaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct atgctagagt
     3601 gcgcactttt tttcctgccg tgtgtataaa gttgcatgta gtcagtcata aacaatagca
     3661 gccagtgtaa taagcattcg ggtaagaacc aactcagttt tttagttgtt tttttttttt
     3721 tttttttcac cccgttgaaa gaaaaggata agagataaag gactaaatga ttttaataca
     3781 gatagacaac cattgttgac aggattgcga ttgtaagagt agacagtaca tcaagcgaaa
     3841 ataaatattg caggaatggt tttgtctgat aaggagttgt ttgccataaa taagaaagcc
     3901 gtcgaacaag gtttcaatgt gaagcctaga ttgaactata atacggtcag tggtgtgaac
     3961 ggtccattag tcattttgga aaaggtcaag ttcccacgtt acaacgaaat tgttaatttg
     4021 acattgccag atggaaccgt gagacaaggt caagttttgg aaattagagg agatagagcc
     4081 attgtgcaag tgtttgaagg tacatctggt attgatgtca agaagactac cgtggaattc
     4141 actggtgaga gtttgagaat tcctgtgtct gaagacatgt tgggtagaat ttttgacggt
     4201 tctggtagac ccattgacaa cggtcctaaa gttttcgcag aggattactt ggacattaac
     4261 ggttctccta tcaacccata tgctcgtatt tatccagaag aaatgatttc tactggtgtt
     4321 tctgctattg acacaatgaa ctccattgcc agaggtcaaa agatcccaat tttctccgca
     4381 tcaggtttac cacacaacga aattgcagca caaatttgta gacaggctgg tttggtgaga
     4441 cctaccaagg atgttcatga tggtcatgaa gaaaatttct ccatcgtttt tgctgccatg
     4501 ggtgtcaact tggaaaccgc tagatttttc aaacaggatt tcgaagaaaa tgggtctttg
     4561 gaaagaactt cattattttt gaacttggct aatgacccta ccattgaaag aattatcact
     4621 ccaagattgg ccttgaccac cgctgaatac cttgcttacc aaacggaacg tcatgtgttg
     4681 accatcttga ccgatatgtc atcgtatgct gatgctctta gagaagtttc cgctgctaga
     4741 gaagaagttc caggtagaag aggttatcct ggttacatgt atacagattt gtccacaatt
     4801 tatgaaagag caggtagagt agagggtcgt aacgggtcca tcactcaaat acctatcttg
     4861 acaatgccta acgatgatat tacgcatcca attccggatt tgaccggtta tattaccgag
     4921 ggtcaaatct tcgttgaccg tcaattacat aacaagggta tctacccacc aatcaacgtc
     4981 ttgccttcgt tgagtagatt gatgaaatct gccatcggtg aaggtatgac cagaaaggac
     5041 cacggtgacg tttctaacca attgtatgcc aagtacgcca tcggtaagga cgctgctgct
     5101 atgaaggccg ttctcgaggg ggggcccggt acccagcttt tgttcccttt agtgagggtt
     5161 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5221 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5281 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5341 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5401 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5461 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5521 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5581 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5641 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5701 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5761 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5821 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5881 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5941 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6001 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6061 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6121 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6181 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6241 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6301 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6361 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6421 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6481 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6541 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6601 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6661 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6721 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6781 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6841 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6901 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6961 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7021 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7081 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7141 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7201 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7261 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7321 ccgaaaagtg ccac
//
