LOCUS       sequence-121958-        7284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of GFP tag at Atg18 C terminus, use auxotrophic marker 
            URA3(Candida albicans).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7284)
  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 7284)
  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: LD118
FEATURES             Location/Qualifiers
     source          1..7284
                     /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             1618..2334
                     /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             1618..2331
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2355..2542
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2410..2434)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2617..2960
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3779..3976
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(5096..5116)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5096..5114)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5135..5151)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5135..5151)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5148..5170)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5159..5175
                     /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(5183..5189,5190..5207,5208..5213))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5228..5249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5366..5383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5537..6125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5617..5636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6296..7087,7088..7156))
                     /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     6919..6938
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7157..7261)
                     /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 ccgcggtcga gctttactaa gagggatgat gcggatccaa caagcagtaa cggcggtaac
      721 agcagtataa taaagaatgg tgatgtgatc gtattcaact tggaaacatt acagccaacc
      781 atggtcatcg aagctcataa gggcgagatt gctgcaatgg caattagttt tgatgggaca
      841 ctaatggcta ccgcctctga taaaggtact atcatcaggg tctttgacat tgaaacgggt
      901 gataagatct accaattcag gagagggacg tacgcgacaa gaatttactc catatcattc
      961 agtgaagata gccagtactt ggcggttacc ggctcttcca aaaccgtgca tatcttcaaa
     1021 ttggggcatt caatgagcaa caataaacta gacagcgatg atagcaacat ggaagaagct
     1081 gcagccgatg attcatcgct cgataccacc agtatcgatg cgctgagtga cgaggaaaac
     1141 ccgacaagac tcgcaagaga accatatgtg gatgcatcaa gaaagacaat gggtaggatg
     1201 atacgttact cttctcaaaa gctatcccga agagctgcca gaacattggg tcagattttc
     1261 cccatcaaag ttacatcgtt gttggaatcc tcgcgccatt ttgcgtcttt gaaacttccc
     1321 gttgaaacca attcccatgt aatgaccata tcaagtatag gctctccaat agatatagac
     1381 acatccgagt atccggaact cttcgaaact ggcaattccg caagtacaga gtcctaccat
     1441 gagcctgtta tgaagatggt ccccatcagg gtcgtttcct cggatggata cctatacaac
     1501 tttgttatgg acccggagag aggcggcgat tgcttaatat tgtcacagta ttccatcttg
     1561 atggatgctt cgtacgctgc aggtcgacgg atcggtgacg gtgctggttt aattaacatg
     1621 tctaaaggtg aagaattatt cactggtgtt gtcccaattt tggttgaatt agatggtgat
     1681 gttaatggtc acaaattttc tgtctccggt gaaggtgaag gtgatgctac ttacggtaaa
     1741 ttgaccttaa aatttatttg tactactggt aaattgccag ttccatggcc aaccttagtc
     1801 actactttaa cttatggtgt tcaatgtttt tctagatacc cagatcatat gaaacaacat
     1861 gactttttca agtctgccat gccagaaggt tatgttcaag aaagaactat ttttttcaaa
     1921 gatgacggta actacaagac cagagctgaa gtcaagtttg aaggtgatac cttagttaat
     1981 agaatcgaat taaaaggtat tgattttaaa gaagatggta acattttagg tcacaaattg
     2041 gaatacaact ataactctca caatgtttac atcatggctg acaaacaaaa gaatggtatc
     2101 aaagttaact tcaaaattag acacaacatt gaagatggtt ctgttcaatt agctgaccat
     2161 tatcaacaaa atactccaat tggtgatggt ccagtcttgt taccagacaa ccattactta
     2221 tccactcaat ctgccttatc caaagatcca aacgaaaaga gagaccacat ggtcttgtta
     2281 gaatttgtta ctgctgctgg tattacccat ggtatggatg aattgtacaa ataaggcgcg
     2341 ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
     2401 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
     2461 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat gaggtcgctc
     2521 ttattgacca cacctctacc ggcagatccg ctagggataa cagggtaata tagatctgtt
     2581 tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc
     2641 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
     2701 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
     2761 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
     2821 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
     2881 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
     2941 acatccgaac ataaacaacc atgacagtca acactaagac ctatagtgag agagcagaaa
     3001 ctcatgcctc accagtagca cagcgattat ttcgattaat ggaactgaag aaaaccaatt
     3061 tatgtgcatc aattgacgtt gataccacta aggaattcct tgaattaatt gataaattag
     3121 gtccttatgt atgcttaatc aagactcata ttgatataat caatgatttt tcctatgaat
     3181 ccactattga accattatta gaactttcac gtaaacatca atttatgatt tttgaagata
     3241 gaaaatttgc tgatattggt aataccgtaa agaaacaata tattggtgga gtttataaaa
     3301 ttagtagttg ggcagatatt accaatgctc atggtgtcac tgggaatgga gtggttgaag
     3361 gattaaaaca gggagctaaa gaaaccacca ccaaccaaga gccaagaggg ttattgatgt
     3421 tagctgaatt atcatcagtg ggatcattag catatggaga atattctcaa aaaactgttg
     3481 aaattgctaa atccgataag gaatttgtta ttggatttat tgcccaacgt gatatgggtg
     3541 gccaagaaga aggatttgat tggcttatta tgacacctgg agttggatta gatgataaag
     3601 gtgatggatt aggacaacaa tatagaactg ttgatgaagt tgttagcact ggaactgata
     3661 ttatcattgt tggtagagga ttgtttggta aaggaagaga tccagatatt gaaggtaaaa
     3721 ggtatagaaa tgctggttgg aatgcttatt tgaaaaagac tggccaatta taatcagtac
     3781 tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt
     3841 agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat
     3901 ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat
     3961 gctggtcgct atactgctgt cgattcgata ctaacgccgc catccagtgt cgaaaacgag
     4021 ctcgaattca tcgatgatat cagatccact agtggcctat gctgatcacg gaagtgtctc
     4081 gcgctgcctt ccgtacgtca caacgcatac acaatatagt ttatataaac tcctgcgtat
     4141 aatctatcct tttgcataat attttcgtca ccattatctt gttgcggtgt ttttatttaa
     4201 aacaatagaa ctcgcctaaa ggggaaattt tcgatataaa aattcaaaaa aatggctttc
     4261 atggatcgag tatttgtttg tcgaaaaagg atcactggag tgacgttact acgctacgaa
     4321 gcctccttcc ggcttagccc tgttgattac gaatttggat cagttggtat ttgattcctc
     4381 ggaagagtta acttcacgcg ggtaaatcac ttgtgatgcg gtatattctc tatacggcta
     4441 atagatgaat cagggtgttt taaagtgcgt ataaaccctt ttgctatttt cgtttatata
     4501 attggcgttg ataaagagcc aatatctatt gttgctacat agaggcagct ctcttagcaa
     4561 aataaaaata caaaaagttc gacatgggct tcagtagcgg taaatcaact aagaaaaagc
     4621 ctctgctttt cgatatcaga cttaaaaatg ttgacaacga tgtaatactc ctcaaaggtc
     4681 ctccaaacga ggccccctcg gtgcttttat ctggttgcat cgttttatcg attaacgaac
     4741 ccatgcagat caaaagcata tcattgagac tttatgggaa gatacaaata gacgtaccat
     4801 tagagaggcc ccaggacgct agttcttcgt cgttgtcttc atcgccgcca aagatcagaa
     4861 agtacaacaa agttttttat aattacgcat gggataatgt taacctcaag gagtatctga
     4921 gtggtttaag agggcaatct ggccttgcgg gcagtagctc atcaagtaat atcttgggca
     4981 ctcgccaaag agctcagtcc acaagttcct tgaagtcttt aaaggggtcc tcctcaccct
     5041 cttcatgtac tttagataag ggctcgaggg ggggcccggt acccagcttt tgttcccttt
     5101 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5161 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5221 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5281 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5341 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5401 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5461 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5521 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5581 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5641 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5701 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5761 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5821 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5881 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5941 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     6001 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6061 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6121 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6181 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6241 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6301 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6361 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6421 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6481 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6541 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6601 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6661 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6721 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6781 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6841 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6901 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6961 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7021 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7081 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7141 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7201 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7261 gcacatttcc ccgaaaagtg ccac
//
