LOCUS       sequence-128243-        6781 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of GFP tag at Atg11 C terminus, use auxotrophic marker 
            URA3(Candida albicans).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6781)
  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 6781)
  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: LD111
FEATURES             Location/Qualifiers
     source          1..6781
                     /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     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1349..2065
                     /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"
     terminator      2086..2273
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2141..2165)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2348..2691
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3510..3707
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(4593..4613)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4593..4611)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4632..4648)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4632..4648)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4645..4667)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4656..4672
                     /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(4680..4686,4687..4704,4705..4710))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4725..4746
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4863..4880)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5034..5622)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5114..5133)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5793..6584,6585..6653))
                     /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     6416..6435
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6654..6758)
                     /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 gccagtgaat tgtaatacga ctcactatag ggcgaattgg agctccaccg
      661 cggtggatct gaaaaggata cacagcaaag catttttctg gggaatatca ctcaattata
      721 tgacaataag ctttatgaag ttgccgttat caggagattc aaggatattg agaccttggc
      781 caaaaaatta actaaagaaa ataagatcaa gagaacgcta ttggagagat tccaaaggga
      841 aaaggtgact ttgaggaact ttcaaattgg cgatttagca ttatttttgc caacacgaga
      901 aaacgttaat tctgtcgggt ctatgtcgtc ttccacctca tcattgagct catcattttc
      961 atctgttgat ttatcaactc caccgccttt ggatgctatg tctatacaat cgtctccctc
     1021 ggtgatacat agtaatgtaa tcaatcaagc cagtatatct ggcagggaca agaataaact
     1081 gatgaggcct tgggctgcat ttacagcttt tgaagaaagc acaagatatt ttttaaagga
     1141 cgaaaaagga ttgacgaagg ggaaagaatg gtttgtaggc agaattgtca ctttggaaca
     1201 tttcgtagca gatagcccct ctaataatcc atttagacta ccaaagggct cagtttggtt
     1261 tcaggtcact gcggttgtgg tttcatacca gggagttgct tcgtacgctg caggtcgacg
     1321 gatcggtgac ggtgctggtt taattaacat gtctaaaggt gaagaattat tcactggtgt
     1381 tgtcccaatt ttggttgaat tagatggtga tgttaatggt cacaaatttt ctgtctccgg
     1441 tgaaggtgaa ggtgatgcta cttacggtaa attgacctta aaatttattt gtactactgg
     1501 taaattgcca gttccatggc caaccttagt cactacttta acttatggtg ttcaatgttt
     1561 ttctagatac ccagatcata tgaaacaaca tgactttttc aagtctgcca tgccagaagg
     1621 ttatgttcaa gaaagaacta tttttttcaa agatgacggt aactacaaga ccagagctga
     1681 agtcaagttt gaaggtgata ccttagttaa tagaatcgaa ttaaaaggta ttgattttaa
     1741 agaagatggt aacattttag gtcacaaatt ggaatacaac tataactctc acaatgttta
     1801 catcatggct gacaaacaaa agaatggtat caaagttaac ttcaaaatta gacacaacat
     1861 tgaagatggt tctgttcaat tagctgacca ttatcaacaa aatactccaa ttggtgatgg
     1921 tccagtcttg ttaccagaca accattactt atccactcaa tctgccttat ccaaagatcc
     1981 aaacgaaaag agagaccaca tggtcttgtt agaatttgtt actgctgctg gtattaccca
     2041 tggtattgat gaattgtaca aataaggcgc gccacttcta aataagcgaa tttcttatga
     2101 tttatgattt ttattattaa ataagttata aaaaaaataa gtgtatacaa attttaaagt
     2161 gactcttagg ttttaaaacg aaaattctta ttcttgagta actctttcct gtaggtcagg
     2221 ttgctttctc aggtatagta tgaggtcgct cttattgacc acacctctac cggcagatcc
     2281 gctagggata acagggtaat atagatctgt ttagcttgcc tcgtccccgc cgggtcaccc
     2341 ggccagcgac atggaggccc agaataccct ccttgacagt cttgacgtgc gcagctcagg
     2401 ggcatgatgt gactgtcgcc cgtacattta gcccatacat ccccatgtat aatcatttgc
     2461 atccatacat tttgatggcc gcacggcgcg aagcaaaaat tacggctcct cgctgcagac
     2521 ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct
     2581 gtagagaaat ataaaaggtt aggatttgcc actgaggttc ttctttcata tacttccttt
     2641 taaaatcttg ctaggataca gttctcacat cacatccgaa cataaacaac catgacagtc
     2701 aacactaaga cctatagtga gagagcagaa actcatgcct caccagtagc acaacgatta
     2761 tttcgattaa tggaactgaa gaaaaccaat ttatgtgcat caattgatgt tgataccact
     2821 aaggaattcc ttgaattaat tgataaattg ggtccttatg tatgcttaat caagactcat
     2881 attgatataa tcaatgattt ttcctatgaa tccactattg aaccattatt agaactttca
     2941 cgtaaacatc aatttatgat ttttgaagat agaaaatttg ctgatattgg taataccgtg
     3001 aagaaacaat atattggtgg agtttataaa attagtagtt gggcagatat tactaatgct
     3061 catggtgtca ctgggaatgg agtagttgaa ggattaaaac agggagctaa agaaaccacc
     3121 accaaccaag agccaagagg gttattgatg ttagctgaat tatcatcagt gggatcatta
     3181 gcatatggag aatattctca aaaaactgtt gaaattgcta aatccgataa ggaatttgtt
     3241 attggattta ttgcccaacg tgatatgggt ggacaagaag aaggatttga ttggcttatt
     3301 atgacacctg gagttggatt agatgataaa ggtgatggat taggacaaca atatagaact
     3361 gttgatgaag ttgttagcac tggaactgat attatcattg ttggtagagg attgtttggt
     3421 aaaggaagag atccagatat tgaaggtaaa aggtatagag atgctggttg gaatgcttat
     3481 ttgaaaaaga ctggccaatt ataatcagta ctgacaataa aaagattctt gttttcaaga
     3541 acttgtcatt tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg
     3601 tgatttatat tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga
     3661 aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat
     3721 actaacgccg ccatccagtg tcgaaaacga gctcgaattc atcgatgata tcagatccac
     3781 tagtggccta tgctgaacag tgctaaacgt ttgtcacaaa tgacaagatt ttaattatgt
     3841 atctatttaa cattataatg cgataagtct tgttatcgac taccaagttt cctggatata
     3901 tctatcagat ttttccattt ctttcaggta ttttttggca gtttcttcat cactaaaatt
     3961 gccccatttt tttaacattt caatgaatgt taatctgact tggataggca ttttacctga
     4021 tgagccacat aagaaaaaaa cggcgtcttt attaactacc aaattagtta tttcctcacc
     4081 taatctccat aagtagtctt ggacatattt aacacctggt gaattttcct catctctgga
     4141 aaaggatgaa tgtaaggcaa ttttgccctt acggaaccaa tcttccagca tatctttata
     4201 aatataatct ttatctttat atctacaacc gaagaggagt ttgatatctt tagatatctc
     4261 cgctttaacc actgataata atggtgctag accgacaccc ggtccgacaa gaatcatagg
     4321 tttattcaaa aattcctttt tgattatgtg attattttgt aacttatacc ttatctgttc
     4381 cccctcttgc aatcgtgcaa tgtagttagt gcaaattcct cttcgaattt tccttaagat
     4441 agttttatat tttactatcg caactgttaa ttcaatatta ggatccccag ggccgctaga
     4501 tatagagtag taccttggtt taatgatggg caagtaatct agcacatatt tccagggcac
     4561 tcgagggggg gcccggtacc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     4621 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4681 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4741 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4801 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4861 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4921 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4981 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5041 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5101 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5161 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5221 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5281 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5341 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5401 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5461 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5521 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5581 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5641 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5701 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5761 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5821 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5881 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5941 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6001 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6061 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6121 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6181 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6241 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6301 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6361 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6421 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6481 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6541 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6601 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6661 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6721 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6781 c
//
