LOCUS       sequence-128290-        8084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of 2GFP 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 8084)
  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 8084)
  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: LD214
FEATURES             Location/Qualifiers
     source          1..8084
                     /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..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"
     CDS             1900..2616
                     /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             1900..2613
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2637..2824
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2692..2716)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2899..3242
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4061..4258
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(5896..5916)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5896..5914)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5935..5951)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5935..5951)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5948..5970)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5959..5975
                     /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(5983..5989,5990..6007,6008..6013))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6028..6049
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6166..6183)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6337..6925)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6417..6436)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7096..7887,7888..7956))
                     /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     7719..7738
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7957..8061)
                     /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 atggtattga tgaattgtac
     1861 aaaattccag ttccaacgcg tgatctagag aagccattca tgtctaaagg tgaagaatta
     1921 ttcactggtg ttgtcccaat tttggttgaa ttagatggtg atgttaatgg tcacaaattt
     1981 tctgtctccg gtgaaggtga aggtgatgct acttacggta aattgacctt aaaatttatt
     2041 tgtactactg gtaaattgcc agttccatgg ccaaccttag tcactacttt aacttatggt
     2101 gttcaatgtt tttctagata cccagatcat atgaaacaac atgacttttt caagtctgcc
     2161 atgccagaag gttatgttca agaaagaact atttttttca aagatgacgg taactacaag
     2221 accagagctg aagtcaagtt tgaaggtgat accttagtta atagaatcga attaaaaggt
     2281 attgatttta aagaagatgg taacatttta ggtcacaaat tggaatacaa ctataactct
     2341 cacaatgttt acatcatggc tgacaaacaa aagaatggta tcaaagttaa cttcaaaatt
     2401 agacacaaca ttgaagatgg ttctgttcaa ttagctgacc attatcaaca aaatactcca
     2461 attggtgatg gtccagtctt gttaccagac aaccattact tatccactca atctgcctta
     2521 tccaaagatc caaacgaaaa gagagaccac atggtcttgt tagaatttgt tactgctgct
     2581 ggtattaccc atggtatgga tgaattatat aaataaggcg cgccacttct aaataagcga
     2641 atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca
     2701 aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc
     2761 tgtaggtcag gttgctttct caggtatagt atgaggtcgc tcttattgac cacacctcta
     2821 ccggcagatc cgctagggat aacagggtaa tatagatctg tttagcttgc ctcgtccccg
     2881 ccgggtcacc cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg
     2941 cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta
     3001 taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc
     3061 tcgctgcaga cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac
     3121 gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat
     3181 atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa
     3241 ccatgacagt caacactaag acctatagtg agagagcaga aactcatgcc tcaccagtag
     3301 cacaacgatt atttcgatta atggaactga agaaaaccaa tttatgtgca tcaattgatg
     3361 ttgataccac taaggaattc cttgaattaa ttgataaatt gggtccttat gtatgcttaa
     3421 tcaagactca tattgatata atcaatgatt tttcctatga atccactatt gaaccattat
     3481 tagaactttc acgtaaacat caatttatga tttttgaaga tagaaaattt gctgatattg
     3541 gtaataccgt gaagaaacaa tatattggtg gagtttataa aattagtagt tgggcagata
     3601 ttactaatgc tcatggtgtc actgggaatg gagtagttga aggattaaaa cagggagcta
     3661 aagaaaccac caccaaccaa gagccaagag ggttattgat gttagctgaa ttatcatcag
     3721 tgggatcatt agcatatgga gaatattctc aaaaaactgt tgaaattgct aaatccgata
     3781 aggaatttgt tattggattt attgcccaac gtgatatggg tggacaagaa gaaggatttg
     3841 attggcttat tatgacacct ggagttggat tagatgataa aggtgatgga ttaggacaac
     3901 aatatagaac tgttgatgaa gttgttagca ctggaactga tattatcatt gttggtagag
     3961 gattgtttgg taaaggaaga gatccagata ttgaaggtaa aaggtataga gatgctggtt
     4021 ggaatgctta tttgaaaaag actggccaat tataatcagt actgacaata aaaagattct
     4081 tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc tattttaatc
     4141 aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag atgcgaagtt
     4201 aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg ctatactgct
     4261 gtcgattcga tactaacgcc gccatccagt gtcgaaaacg agctcgaatt catcgatgat
     4321 atcagatcca ctagtggcct atgctagagt gcgcactttt tttcctgccg tgtgtataaa
     4381 gttgcatgta gtcagtcata aacaatagca gccagtgtaa taagcattcg ggtaagaacc
     4441 aactcagttt tttagttgtt tttttttttt tttttttcac cccgttgaaa gaaaaggata
     4501 agagataaag gactaaatga ttttaataca gatagacaac cattgttgac aggattgcga
     4561 ttgtaagagt agacagtaca tcaagcgaaa ataaatattg caggaatggt tttgtctgat
     4621 aaggagttgt ttgccataaa taagaaagcc gtcgaacaag gtttcaatgt gaagcctaga
     4681 ttgaactata atacggtcag tggtgtgaac ggtccattag tcattttgga aaaggtcaag
     4741 ttcccacgtt acaacgaaat tgttaatttg acattgccag atggaaccgt gagacaaggt
     4801 caagttttgg aaattagagg agatagagcc attgtgcaag tgtttgaagg tacatctggt
     4861 attgatgtca agaagactac cgtggaattc actggtgaga gtttgagaat tcctgtgtct
     4921 gaagacatgt tgggtagaat ttttgacggt tctggtagac ccattgacaa cggtcctaaa
     4981 gttttcgcag aggattactt ggacattaac ggttctccta tcaacccata tgctcgtatt
     5041 tatccagaag aaatgatttc tactggtgtt tctgctattg acacaatgaa ctccattgcc
     5101 agaggtcaaa agatcccaat tttctccgca tcaggtttac cacacaacga aattgcagca
     5161 caaatttgta gacaggctgg tttggtgaga cctaccaagg atgttcatga tggtcatgaa
     5221 gaaaatttct ccatcgtttt tgctgccatg ggtgtcaact tggaaaccgc tagatttttc
     5281 aaacaggatt tcgaagaaaa tgggtctttg gaaagaactt cattattttt gaacttggct
     5341 aatgacccta ccattgaaag aattatcact ccaagattgg ccttgaccac cgctgaatac
     5401 cttgcttacc aaacggaacg tcatgtgttg accatcttga ccgatatgtc atcgtatgct
     5461 gatgctctta gagaagtttc cgctgctaga gaagaagttc caggtagaag aggttatcct
     5521 ggttacatgt atacagattt gtccacaatt tatgaaagag caggtagagt agagggtcgt
     5581 aacgggtcca tcactcaaat acctatcttg acaatgccta acgatgatat tacgcatcca
     5641 attccggatt tgaccggtta tattaccgag ggtcaaatct tcgttgaccg tcaattacat
     5701 aacaagggta tctacccacc aatcaacgtc ttgccttcgt tgagtagatt gatgaaatct
     5761 gccatcggtg aaggtatgac cagaaaggac cacggtgacg tttctaacca attgtatgcc
     5821 aagtacgcca tcggtaagga cgctgctgct atgaaggccg ttctcgaggg ggggcccggt
     5881 acccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     5941 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6001 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6061 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6121 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6181 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6241 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6301 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6361 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6421 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6481 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6541 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6601 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6661 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6721 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6781 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6841 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6901 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6961 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7021 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7081 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7141 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7201 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7261 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7321 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7381 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7441 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7501 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7561 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7621 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7681 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7741 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7801 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7861 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7921 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7981 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8041 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccac
//
