LOCUS       sequence-121949-        7237 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of GFP tag at Atg2 C terminus, use auxotrophic marker 
            URA3(Candida albicans).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7237)
  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 7237)
  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: LD102
FEATURES             Location/Qualifiers
     source          1..7237
                     /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"
     primer_bind     677..693
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     677..693
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(727..743)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(728..744)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             1973..2689
                     /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             1973..2686
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2710..2897
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2765..2789)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2972..3315
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4134..4331
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(5049..5069)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5049..5067)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5088..5104)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5088..5104)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5101..5123)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5112..5128
                     /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(5136..5142,5143..5160,5161..5166))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5181..5202
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5319..5336)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5490..6078)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5570..5589)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6249..7040,7041..7109))
                     /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     6872..6891
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7110..7214)
                     /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 ccgcggtggc ggccgctcta gaactagtgg atcccccggg ctgcaggaat tcgatatcaa
      721 gcttatcgat accgtcgacc tcgagtttct tctggacctg gaatttatcc ggccgatcga
      781 ctttttgcaa gcagtggaat tagtcatgca attaaatgtt gcgcctttac gccttcacgt
      841 ggatcaagac actttagagt ttctaataag gttcttaggg ttcaaagata aaagatttga
      901 attaattgat gagtatccag atattgtatt tattcaaaaa tttagcacta attcaattaa
      961 gttaagatta gattataagc cgaaaaaagt tgattatgcg ggtttaagat cgggccaaac
     1021 ttctgaattg atgaattttt ttacccttga tggatccaag attattttga aaagcgtagt
     1081 cctgtatgga ttaaatggat tcgatgaatt aaataataaa ctgaaagcca tttggacgcc
     1141 ggatattacc aaaaagcagc tgcctggcgt tttggaagga ctggctcctg tcagatcgtt
     1201 tatggcgatt gggtcagggg tgaagaccct agtcacagta ctaatgtcag aatacaggca
     1261 agaagggcat ttgggaagga gtctgaaaaa gggtggcaat gtcttcttga aaacgacaac
     1321 gggagatttt gtgaaattgg gagttaaatt aacttcaggg acacaggcga tattggagaa
     1381 caccgaggaa ctattcggtg gtgttggttc caatggcagg gtgtacgatg catcgaaatt
     1441 tggctccgct gatggtgcag acagtgatac tgcagctgta ttagacttgg atactttatt
     1501 cgaggaggat cagttagttg gtagtaaata ttccaggata agagatcatg agcccacggc
     1561 agtagtcatc gatatgtctt caccgggaga tcacaacgaa cccacaatag tgagtctgta
     1621 cgcagatcag cccctcgacc tgcccacggg tttgaaggag gcgtatagtt cattagaaaa
     1681 gcatatgcat attgcgtacg atgcggtatg gagggcaaag ggacagatga aggatgataa
     1741 acggggaggg ccgagcgccg cagcagttta tgttgctaga gctgcacctg tggcaattat
     1801 tcggccgtta attggggcca ctgaggcagt gtctaagact ttacagggaa tagctaatca
     1861 agtggataag acgcataacg agcaaatcaa tgataagtac aagtccaatc ggactgattc
     1921 ggcttcgtac gctgcaggtc gacggatcgg tgacggtgct ggtttaatta acatgtctaa
     1981 aggtgaagaa ttattcactg gtgttgtccc aattttggtt gaattagatg gtgatgttaa
     2041 tggtcacaaa ttttctgtct ccggtgaagg tgaaggtgat gctacttacg gtaaattgac
     2101 cttaaaattt atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac
     2161 tttaacttat ggtgttcaat gtttttctag atacccagat catatgaaac aacatgactt
     2221 tttcaagtct gccatgccag aaggttatgt tcaagaaaga actatttttt tcaaagatga
     2281 cggtaactac aagaccagag ctgaagtcaa gtttgaaggt gataccttag ttaatagaat
     2341 cgaattaaaa ggtattgatt ttaaagaaga tggtaacatt ttaggtcaca aattggaata
     2401 caactataac tctcacaatg tttacatcat ggctgacaaa caaaagaatg gtatcaaagt
     2461 taacttcaaa attagacaca acattgaaga tggttctgtt caattagctg accattatca
     2521 acaaaatact ccaattggtg atggtccagt cttgttacca gacaaccatt acttatccac
     2581 tcaatctgcc ttatccaaag atccaaacga aaagagagac cacatggtct tgttagaatt
     2641 tgttactgct gctggtatta cccatggtat ggatgaattg tacaaataag gcgcgccact
     2701 tctaaataag cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa
     2761 ataagtgtat acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg
     2821 agtaactctt tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt
     2881 gaccacacct ctaccggcag atccgctagg gataacaggg taatatagat ctgtttagct
     2941 tgcctcgtcc ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga
     3001 cagtcttgac gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat
     3061 acatccccat gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa
     3121 aaattacggc tcctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt
     3181 gaattgtccc cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag
     3241 gttcttcttt catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc
     3301 cgaacataaa caaccatgac agtcaacact aagacctata gtgagagagc agaaactcat
     3361 gcctcaccag tagcacagcg attatttcga ttaatggaac tgaagaaaac caatttatgt
     3421 gcatcaattg acgttgatac cactaaggaa ttccttgaat taattgataa attaggtcct
     3481 tatgtatgct taatcaagac tcatattgat ataatcaatg atttttccta tgaatccact
     3541 attgaaccat tattagaact ttcacgtaaa catcaattta tgatttttga agatagaaaa
     3601 tttgctgata ttggtaatac cgtaaagaaa caatatattg gtggagttta taaaattagt
     3661 agttgggcag atattaccaa tgctcatggt gtcactggga atggagtggt tgaaggatta
     3721 aaacagggag ctaaagaaac caccaccaac caagagccaa gagggttatt gatgttagct
     3781 gaattatcat cagtgggatc attagcatat ggagaatatt ctcaaaaaac tgttgaaatt
     3841 gctaaatccg ataaggaatt tgttattgga tttattgccc aacgtgatat gggtggccaa
     3901 gaagaaggat ttgattggct tattatgaca cctggagttg gattagatga taaaggtgat
     3961 ggattaggac aacaatatag aactgttgat gaagttgtta gcactggaac tgatattatc
     4021 attgttggta gaggattgtt tggtaaagga agagatccag atattgaagg taaaaggtat
     4081 agaaatgctg gttggaatgc ttatttgaaa aagactggcc aattataatc agtactgaca
     4141 ataaaaagat tcttgttttc aagaacttgt catttgtata gtttttttat attgtagttg
     4201 ttctatttta atcaaatgtt agcgtgattt atattttttt tcgcctcgac atcatctgcc
     4261 cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg
     4321 tcgctatact gctgtcgatt cgatactaac gccgccatcc agtgtcgaaa acgagctcga
     4381 attcatcgat gatatcagat ccactagtgg cctatgctaa aaatttttgc agacattttt
     4441 gatatatatt caattcatat tttatctctt tttttttttt ttttttcaga cgtggaatgg
     4501 tgggaaagcc aaaatgtcac tgaccgcggc taaaagttaa aaaatacatg agacgacatt
     4561 cttttatgta tatatattta cacatgtata caaacaaata cacacgcaag tagagaggag
     4621 ttggtggggg gaagatgcca ttatagagga gaaaagaaca aagagagtga gcttgcaaga
     4681 taaaatcact cgaaaaaaaa aatttcagtg acttagccga taaatgaatg tgcttgcatt
     4741 tttctaatta tccttcgtat cttctggctt agtcacgggc caagcgtaag ggtgcttttc
     4801 gggcataaca tacttgtgtt tttgcatata ttccttcaat ccctttggac ctcttgatcc
     4861 gtaggggtaa atttccggtg ttggaccgtc cggacgctct atgtgcttca gtaatggggt
     4921 gaatatgccc caactgatat ccaattcgtc atctctgaca aagttggaat ggtcacccag
     4981 tagggcgtct cttatcaaca cctcgtaagc ctctggaatc caaaagtctt ggtacccagc
     5041 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     5101 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5161 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5221 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5281 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5341 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5401 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5461 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5521 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5581 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5641 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5701 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5761 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5821 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5881 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5941 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6001 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6061 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6121 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6181 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6241 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6301 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6361 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6421 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6481 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6541 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6601 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6661 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6721 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6781 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6841 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6901 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6961 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7021 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7081 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7141 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7201 ataggggttc cgcgcacatt tccccgaaaa gtgccac
//
