LOCUS       sequence-128249-        6684 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of GFP tag at Atg23 C terminus, use auxotrophic marker 
            URA3(Candida albicans).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6684)
  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 6684)
  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: LD123
FEATURES             Location/Qualifiers
     source          1..6684
                     /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             1277..1993
                     /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      2014..2201
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2069..2093)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2276..2619
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3438..3635
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(4496..4516)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4496..4514)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4535..4551)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4535..4551)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4548..4570)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4559..4575
                     /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(4583..4589,4590..4607,4608..4613))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4628..4649
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4766..4783)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4937..5525)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5017..5036)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5696..6487,6488..6556))
                     /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     6319..6338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6557..6661)
                     /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 ccgcggaact atcaacatga aaaattttgt tcatatgaag gatctcatag aactgaaaat
      721 tgaagatcta caagagcaac taatgagaaa taaaaatgag tcttctacag tactaacgca
      781 acgtgaatta tggctagatt gtcaaaaaaa agtaggagat ttggaaagca aattaataac
      841 caaacttcgg agcagcagca actccaaaat acctccaaat gaaatgtcag aaatgatcaa
      901 ttctacaata cagtacctaa ataaccttct tgactcttct gatgaaaagc tcacgacaac
      961 tttaatatca aacgaaaggg acgtactttc gaaagcttgt gaagaactcc atagcgaaag
     1021 taccactgca caagatgggt caagcgcact accttctaaa ccaattgata tacacaaaag
     1081 ccataaaggg tcgaacgcga gttcaaattt aaaacagccc tcgactccct cattcttggt
     1141 tgctagcaaa tcgccaccaa agattggcat ttctgaaagt gtagtaaacg ctaataaaaa
     1201 tgatgccata tcaaaaaaag ttgaagcttc gtacgctgca ggtcgacgga tcggtgacgg
     1261 tgctggttta attaacatgt ctaaaggtga agaattattc actggtgttg tcccaatttt
     1321 ggttgaatta gatggtgatg ttaatggtca caaattttct gtctccggtg aaggtgaagg
     1381 tgatgctact tacggtaaat tgaccttaaa atttatttgt actactggta aattgccagt
     1441 tccatggcca accttagtca ctactttaac ttatggtgtt caatgttttt ctagataccc
     1501 agatcatatg aaacaacatg actttttcaa gtctgccatg ccagaaggtt atgttcaaga
     1561 aagaactatt tttttcaaag atgacggtaa ctacaagacc agagctgaag tcaagtttga
     1621 aggtgatacc ttagttaata gaatcgaatt aaaaggtatt gattttaaag aagatggtaa
     1681 cattttaggt cacaaattgg aatacaacta taactctcac aatgtttaca tcatggctga
     1741 caaacaaaag aatggtatca aagttaactt caaaattaga cacaacattg aagatggttc
     1801 tgttcaatta gctgaccatt atcaacaaaa tactccaatt ggtgatggtc cagtcttgtt
     1861 accagacaac cattacttat ccactcaatc tgccttatcc aaagatccaa acgaaaagag
     1921 agaccacatg gtcttgttag aatttgttac tgctgctggt attacccatg gtattgatga
     1981 attgtacaaa taaggcgcgc cacttctaaa taagcgaatt tcttatgatt tatgattttt
     2041 attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt
     2101 ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag
     2161 gtatagtatg aggtcgctct tattgaccac acctctaccg gcagatccgc tagggataac
     2221 agggtaatat agatctgttt agcttgcctc gtccccgccg ggtcacccgg ccagcgacat
     2281 ggaggcccag aataccctcc ttgacagtct tgacgtgcgc agctcagggg catgatgtga
     2341 ctgtcgcccg tacatttagc ccatacatcc ccatgtataa tcatttgcat ccatacattt
     2401 tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct gcgagcaggg
     2461 aaacgctccc ctcacagacg cgttgaattg tccccacgcc gcgcccctgt agagaaatat
     2521 aaaaggttag gatttgccac tgaggttctt ctttcatata cttcctttta aaatcttgct
     2581 aggatacagt tctcacatca catccgaaca taaacaacca tgacagtcaa cactaagacc
     2641 tatagtgaga gagcagaaac tcatgcctca ccagtagcac agcgattatt tcgattaatg
     2701 gaactgaaga aaaccaattt atgtgcatca attgacgttg ataccactaa ggaattcctt
     2761 gaattaattg ataaattagg tccttatgta tgcttaatca agactcatat tgatataatc
     2821 aatgattttt cctatgaatc cactattgaa ccattattag aactttcacg taaacatcaa
     2881 tttatgattt ttgaagatag aaaatttgct gatattggta ataccgtaaa gaaacaatat
     2941 attggtggag tttataaaat tagtagttgg gcagatatta ccaatgctca tggtgtcact
     3001 gggaatggag tggttgaagg attaaaacag ggagctaaag aaaccaccac caaccaagag
     3061 ccaagagggt tattgatgtt agctgaatta tcatcagtgg gatcattagc atatggagaa
     3121 tattctcaaa aaactgttga aattgctaaa tccgataagg aatttgttat tggatttatt
     3181 gcccaacgtg atatgggtgg ccaagaagaa ggatttgatt ggcttattat gacacctgga
     3241 gttggattag atgataaagg tgatggatta ggacaacaat atagaactgt tgatgaagtt
     3301 gttagcactg gaactgatat tatcattgtt ggtagaggat tgtttggtaa aggaagagat
     3361 ccagatattg aaggtaaaag gtatagaaat gctggttgga atgcttattt gaaaaagact
     3421 ggccaattat aatcagtact gacaataaaa agattcttgt tttcaagaac ttgtcatttg
     3481 tatagttttt ttatattgta gttgttctat tttaatcaaa tgttagcgtg atttatattt
     3541 tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat
     3601 gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc gattcgatac taacgccgcc
     3661 atccagtgtc gaaaacgagc tcgaattcat cgatgatatc agatccacta gtggcctatg
     3721 ctgaaatgca aatagctaga gtagccatga ggataatgta aatttaaaca ttgtatgccc
     3781 acgtaatagt caaactcttt cacgtcaaat tgaaaaaaaa tgttcttttt gctgaaataa
     3841 tgaaaaataa tgaaaaataa tgttgctatt aaattcaata taaatgtata caccaatata
     3901 tatgcaggta taattcctaa cacttttact tcaagatcat gatctaggct ttcttttttt
     3961 tgggataaat gggctagatg aagcattccg tctcgaactg ttgcgtgtgc ctgtgggccc
     4021 actaccaggt cctttgaatg ctgaagattg tggttgcgga tgggggttta aatcatatgg
     4081 atcttctgcc tcagcaggat agctgtcttg cggatgggcg ggcagagttt catgcagttc
     4141 gcctgtaccc acaggagccg ttgagtattg attcgcatta cccacatgtt gagagagggt
     4201 cctatttgct ccattctgtc cattttgatt gggttcaccg taatcggaag cagtcaggat
     4261 cctcgaagaa ccaccggata aaacactatt tgaacttctg tgttttgtaa tagcagcagg
     4321 tacctgcgtc tgctttgctt gcgaaacttg tggatcgtcg tatatactag cgctgaaatt
     4381 ttggttatga gaagccaccc tgttgttacg accatttgct tcacttttca ttaatgaaga
     4441 atcggagatt gacggcgtat aactcgaggg ggggcccggt acccagcttt tgttcccttt
     4501 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4561 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4621 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4681 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4741 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4801 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4861 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4921 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4981 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5041 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5101 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5161 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5221 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5281 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5341 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     5401 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5461 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5521 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5581 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5641 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5701 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5761 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5821 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5881 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5941 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6001 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6061 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6121 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6181 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6241 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6301 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6361 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6421 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6481 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6541 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6601 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6661 gcacatttcc ccgaaaagtg ccac
//
