LOCUS       sequence-121964-        7688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of GFP tag at Atg29 C terminus, use auxotrophic marker 
            URA3(Candida albicans).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7688)
  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 7688)
  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: LD129
FEATURES             Location/Qualifiers
     source          1..7688
                     /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             2207..2923
                     /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             2207..2920
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2944..3131
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2999..3023)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        3206..3549
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4368..4565
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(5500..5520)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5500..5518)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5539..5555)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5539..5555)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5552..5574)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5563..5579
                     /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(5587..5593,5594..5611,5612..5617))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5632..5653
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5770..5787)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5941..6529)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6021..6040)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6700..7491,7492..7560))
                     /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     7323..7342
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7561..7665)
                     /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 ccgcggccaa ataagttaaa atctgcggtc cattgtagca agtaaggaat atgtgattca
      721 taaatttcga atttctttcc gaaaatcttg ccatctctaa tcaattccga aatcctactt
      781 agacaagaag gattaagcaa agaaattttg taaaacagat tccagccaac gtggtaacca
      841 taaaatggta taccctttac aacagagaca tcagcgacaa aattgagatt tccaagtttg
      901 tcgccggcta aatcgtgttt atcatctttt ttccttttaa aggatgccct aatttttact
      961 tccagcgttt tatgaacttg ggcacatctt tggtgtctca atgtactcgt gtcagttatt
     1021 tgtccatcat acttgatgaa catgtaaggt agaatgccat ggacgtggca caatacttga
     1081 tgccctgtgg gtaaagcacc aaacaccctg atgtttggga cctgtgaaaa ctggtttaag
     1141 ggcaggcttt caccgtgcga tgggtccaaa aaggtgggtt ttgacatata gtagtcttga
     1201 ttatttagct ggattctaaa atagtctgat ttagaggatg atctaaccgt atcgctctgt
     1261 attgtgtcgt tcgactccct cgacatttcc aaacatttta tttcgccaca acttgtagtt
     1321 ttgtattgac tcaaatactt tctcttgttt tgtgcttcta ttacagagat ggcttttttc
     1381 ttcaagtaca ctatttctag acgcactgca ctcgtcaaaa aaatgattca aggtgaaaat
     1441 aaaggggtgt tagtgaatac ttatcaagaa gagacataca taattgtact gctgtgcatt
     1501 ttcctacttg actttcatga ttatgaatag tacaaacaca gttgtatata tcaaagttaa
     1561 gggtaggaga ccacagggat tcttggatcc tcccaaattt gagtggaatg gaacaaagga
     1621 gcgacagctt tggacaatgg tatcaaattt gaattattca caagatcaaa tagattggca
     1681 gaatttatca aaaatatttg aaacgcctga attttttctt aaaaagagaa catataaatt
     1741 atttgctgaa cacttagagc ttttacaact acaattggag aaaaaaagag atcttgagaa
     1801 gtattcaaat gatcaagtga atgaagggat gtctgattta atacataaat atacccctac
     1861 tttacaaaat gataatctat taaatgtatc cgcaagccca ctgaccacgg aaagacagga
     1921 ttctgaagaa gttgagacag aagtaacaaa tgaggcgtta caacatttgc aaacttccaa
     1981 aattttgaac attcacaaga aaacatcgga tagtgagaat aagcctaatg acaaattgga
     2041 taaggatggt attaataaag aaatggagtg cggtagttca gatgacgatt tatcttcgag
     2101 cttaagtgtt agtaaatctg cgttggaaga agcgctaatg gacagattgc aattcgcttc
     2161 gtacgctgca ggtcgacgga tcggtgacgg tgctggttta attaacatgt ctaaaggtga
     2221 agaattattc actggtgttg tcccaatttt ggttgaatta gatggtgatg ttaatggtca
     2281 caaattttct gtctccggtg aaggtgaagg tgatgctact tacggtaaat tgaccttaaa
     2341 atttatttgt actactggta aattgccagt tccatggcca accttagtca ctactttaac
     2401 ttatggtgtt caatgttttt ctagataccc agatcatatg aaacaacatg actttttcaa
     2461 gtctgccatg ccagaaggtt atgttcaaga aagaactatt tttttcaaag atgacggtaa
     2521 ctacaagacc agagctgaag tcaagtttga aggtgatacc ttagttaata gaatcgaatt
     2581 aaaaggtatt gattttaaag aagatggtaa cattttaggt cacaaattgg aatacaacta
     2641 taactctcac aatgtttaca tcatggctga caaacaaaag aatggtatca aagttaactt
     2701 caaaattaga cacaacattg aagatggttc tgttcaatta gctgaccatt atcaacaaaa
     2761 tactccaatt ggtgatggtc cagtcttgtt accagacaac cattacttat ccactcaatc
     2821 tgccttatcc aaagatccaa acgaaaagag agaccacatg gtcttgttag aatttgttac
     2881 tgctgctggt attacccatg gtatggatga attgtacaaa taaggcgcgc cacttctaaa
     2941 taagcgaatt tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt
     3001 gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac
     3061 tctttcctgt aggtcaggtt gctttctcag gtatagtatg aggtcgctct tattgaccac
     3121 acctctaccg gcagatccgc tagggataac agggtaatat agatctgttt agcttgcctc
     3181 gtccccgccg ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct
     3241 tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc
     3301 ccatgtataa tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta
     3361 cggctcctcg ctgcagacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg
     3421 tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt
     3481 ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca
     3541 taaacaacca tgacagtcaa cactaagacc tatagtgaga gagcagaaac tcatgcctca
     3601 ccagtagcac agcgattatt tcgattaatg gaactgaaga aaaccaattt atgtgcatca
     3661 attgacgttg ataccactaa ggaattcctt gaattaattg ataaattagg tccttatgta
     3721 tgcttaatca agactcatat tgatataatc aatgattttt cctatgaatc cactattgaa
     3781 ccattattag aactttcacg taaacatcaa tttatgattt ttgaagatag aaaatttgct
     3841 gatattggta ataccgtaaa gaaacaatat attggtggag tttataaaat tagtagttgg
     3901 gcagatatta ccaatgctca tggtgtcact gggaatggag tggttgaagg attaaaacag
     3961 ggagctaaag aaaccaccac caaccaagag ccaagagggt tattgatgtt agctgaatta
     4021 tcatcagtgg gatcattagc atatggagaa tattctcaaa aaactgttga aattgctaaa
     4081 tccgataagg aatttgttat tggatttatt gcccaacgtg atatgggtgg ccaagaagaa
     4141 ggatttgatt ggcttattat gacacctgga gttggattag atgataaagg tgatggatta
     4201 ggacaacaat atagaactgt tgatgaagtt gttagcactg gaactgatat tatcattgtt
     4261 ggtagaggat tgtttggtaa aggaagagat ccagatattg aaggtaaaag gtatagaaat
     4321 gctggttgga atgcttattt gaaaaagact ggccaattat aatcagtact gacaataaaa
     4381 agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta gttgttctat
     4441 tttaatcaaa tgttagcgtg atttatattt tttttcgcct cgacatcatc tgcccagatg
     4501 cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta
     4561 tactgctgtc gattcgatac taacgccgcc atccagtgtc gaaaacgagc tcgaattcat
     4621 cgatgatatc agatccacta gtggcctatg ctttacatct ttcacaagca caatcaaaaa
     4681 accaactgtc ggccaaaaag gcccggcgtt ttactgttgg taagtccagt accccggaat
     4741 aatcgataca gatctgctca tcctttttta tatctctatt catagtgaat agcattgaat
     4801 tacctttacg atatttcgtt atattggggt tacaagagtg gttgaaatag gaggcttctg
     4861 gaaataccca gtatccaaaa tattctctgc tgtctgaggc ttcgccctct tgccataaac
     4921 caaaagcatt cccatattca gtacctaata tatgtctcaa tagcggtatg gagagcattc
     4981 tatgcaaatg tgaaggtaat agaatataga gtgtttgaaa tactaatttt tgaaagtgca
     5041 gtaagacagg aaatttagat attttgctaa gctcattcga ttgtaacata ttgaaagctc
     5101 ggtatgttat gcactgagga tccatatatt taagattaaa caaactctcg cacacaaacc
     5161 taatgcagca atattcatct tcgcaagtgg gtggtaattg atttattctc ttcgcagatt
     5221 tcatgttgtt gatgcgcggt atccatttac tttcaatctc atcccatgaa gactgtatga
     5281 cattttccga aattaagata gagtttaatt tttcctcctg ttcagaggta taattatatc
     5341 ttttcagcat gcttggaaaa tgatgcagta atatctcata acattcaatt aattcaatga
     5401 tatttggtat ttgaagataa gaagttctgc aatgttctga acagaaccac agtccagcgc
     5461 ccagaactcg agggggggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc
     5521 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5581 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5641 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5701 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5761 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5821 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5881 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5941 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6001 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6061 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6121 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6181 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6241 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6301 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6361 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     6421 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6481 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6541 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6601 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6661 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6721 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6781 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6841 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6901 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6961 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7021 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7081 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7141 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7201 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7261 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7321 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7381 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7441 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7501 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7561 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7621 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7681 agtgccac
//
