LOCUS       sequence-128289-        7945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of 2GFP tag at Atg13 C terminus, use auxotrophic marker 
            URA3(Candida albicans).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7945)
  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 7945)
  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: LD213
FEATURES             Location/Qualifiers
     source          1..7945
                     /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             1432..2145
                     /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"
     CDS             2182..2898
                     /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             2182..2895
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2919..3106
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2974..2998)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        3181..3524
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4343..4540
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(5711..5727)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5712..5728)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5757..5777)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5757..5775)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5796..5812)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5796..5812)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5809..5831)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5820..5836
                     /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(5844..5850,5851..5868,5869..5874))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5889..5910
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6027..6044)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6198..6786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6278..6297)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6957..7748,7749..7817))
                     /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     7580..7599
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7818..7922)
                     /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 ccgcggcaga atttgaagcc aagtaatgac ttattaagtg aagatttatc cgtaagttta
      721 tccatggatc caaatcatac atatcacaga ggcagatcag attcccactc accattgcct
      781 tcaatatccc cttcgatgca ttatggatcg ttgaactcga gaatgtctca aggcgccaat
      841 gcaagccatt tgattgcaag aggcggtggg aattcatcta ctagtgcctt gaatagtaga
      901 aggaattctt tagataagag ctcaaacaag cagggtatgt caggcttacc tcctattttt
      961 ggtggagaga gtacttcata tcaccacgac aacaaaatac aaaagtacaa ccaattagga
     1021 gtagaagaag atgatgatga cgagaatgac cgtttgctca accaaatggg aaacagtgct
     1081 acaaaattca aaagttcaat atctccaaga tcaattgata gcatttcaag ttctttcata
     1141 aaaagtagga tacctatcag acaaccatac cattactctc aaccaactac tgcgcccttt
     1201 caagctcagg cgaaatttca taaacctgca aataagttaa tcgataatgg taataggagt
     1261 aatagtaaca ataacaatca taatgggaat gatgcagttg gtgtgatgca taatgacgag
     1321 gatgatcaag atgatgatct agtatttttc atgagtgata tgaacctttc taaagaaggt
     1381 gcttcgtacg ctgcaggtcg acggatcggt gacggtgctg gtttaattaa catgtctaaa
     1441 ggtgaagaat tattcactgg tgttgtccca attttggttg aattagatgg tgatgttaat
     1501 ggtcacaaat tttctgtctc cggtgaaggt gaaggtgatg ctacttacgg taaattgacc
     1561 ttaaaattta tttgtactac tggtaaattg ccagttccat ggccaacctt agtcactact
     1621 ttaacttatg gtgttcaatg tttttctaga tacccagatc atatgaaaca acatgacttt
     1681 ttcaagtctg ccatgccaga aggttatgtt caagaaagaa ctattttttt caaagatgac
     1741 ggtaactaca agaccagagc tgaagtcaag tttgaaggtg ataccttagt taatagaatc
     1801 gaattaaaag gtattgattt taaagaagat ggtaacattt taggtcacaa attggaatac
     1861 aactataact ctcacaatgt ttacatcatg gctgacaaac aaaagaatgg tatcaaagtt
     1921 aacttcaaaa ttagacacaa cattgaagat ggttctgttc aattagctga ccattatcaa
     1981 caaaatactc caattggtga tggtccagtc ttgttaccag acaaccatta cttatccact
     2041 caatctgcct tatccaaaga tccaaacgaa aagagagacc acatggtctt gttagaattt
     2101 gttactgctg ctggtattac ccatggtatt gatgaattgt acaaaattcc agttccaacg
     2161 cgtgatctag agaagccatt catgtctaaa ggtgaagaat tattcactgg tgttgtccca
     2221 attttggttg aattagatgg tgatgttaat ggtcacaaat tttctgtctc cggtgaaggt
     2281 gaaggtgatg ctacttacgg taaattgacc ttaaaattta tttgtactac tggtaaattg
     2341 ccagttccat ggccaacctt agtcactact ttaacttatg gtgttcaatg tttttctaga
     2401 tacccagatc atatgaaaca acatgacttt ttcaagtctg ccatgccaga aggttatgtt
     2461 caagaaagaa ctattttttt caaagatgac ggtaactaca agaccagagc tgaagtcaag
     2521 tttgaaggtg ataccttagt taatagaatc gaattaaaag gtattgattt taaagaagat
     2581 ggtaacattt taggtcacaa attggaatac aactataact ctcacaatgt ttacatcatg
     2641 gctgacaaac aaaagaatgg tatcaaagtt aacttcaaaa ttagacacaa cattgaagat
     2701 ggttctgttc aattagctga ccattatcaa caaaatactc caattggtga tggtccagtc
     2761 ttgttaccag acaaccatta cttatccact caatctgcct tatccaaaga tccaaacgaa
     2821 aagagagacc acatggtctt gttagaattt gttactgctg ctggtattac ccatggtatg
     2881 gatgaattat ataaataagg cgcgccactt ctaaataagc gaatttctta tgatttatga
     2941 tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt
     3001 aggttttaaa acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt
     3061 ctcaggtata gtatgaggtc gctcttattg accacacctc taccggcaga tccgctaggg
     3121 ataacagggt aatatagatc tgtttagctt gcctcgtccc cgccgggtca cccggccagc
     3181 gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga
     3241 tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata
     3301 cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag
     3361 cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga
     3421 aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc
     3481 ttgctaggat acagttctca catcacatcc gaacataaac aaccatgaca gtcaacacta
     3541 agacctatag tgagagagca gaaactcatg cctcaccagt agcacagcga ttatttcgat
     3601 taatggaact gaagaaaacc aatttatgtg catcaattga cgttgatacc actaaggaat
     3661 tccttgaatt aattgataaa ttaggtcctt atgtatgctt aatcaagact catattgata
     3721 taatcaatga tttttcctat gaatccacta ttgaaccatt attagaactt tcacgtaaac
     3781 atcaatttat gatttttgaa gatagaaaat ttgctgatat tggtaatacc gtaaagaaac
     3841 aatatattgg tggagtttat aaaattagta gttgggcaga tattaccaat gctcatggtg
     3901 tcactgggaa tggagtggtt gaaggattaa aacagggagc taaagaaacc accaccaacc
     3961 aagagccaag agggttattg atgttagctg aattatcatc agtgggatca ttagcatatg
     4021 gagaatattc tcaaaaaact gttgaaattg ctaaatccga taaggaattt gttattggat
     4081 ttattgccca acgtgatatg ggtggccaag aagaaggatt tgattggctt attatgacac
     4141 ctggagttgg attagatgat aaaggtgatg gattaggaca acaatataga actgttgatg
     4201 aagttgttag cactggaact gatattatca ttgttggtag aggattgttt ggtaaaggaa
     4261 gagatccaga tattgaaggt aaaaggtata gaaatgctgg ttggaatgct tatttgaaaa
     4321 agactggcca attataatca gtactgacaa taaaaagatt cttgttttca agaacttgtc
     4381 atttgtatag tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta
     4441 tatttttttt cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat
     4501 atcatgcgtc aatcgtatgt gaatgctggt cgctatactg ctgtcgattc gatactaacg
     4561 ccgccatcca gtgtcgaaaa cgagctcgaa ttcatcgatg atatcagatc cactagtggc
     4621 ctatgctaaa aatggtaaag ttttatttta aagggcacaa ctaaagaaaa ataatcaaaa
     4681 aaaaggtgaa atagatatgc aagaaaagaa aaaaaaatac aatactagta aactgaaata
     4741 aaaaagatta gccataatgg atttgtttct ctttgttatc gatattctta cctcttgaag
     4801 atccatgtgt ctatcatttg aattgaatat ataattatat acagtgagat ataccaattg
     4861 cagtaacaaa atggcaaata tgtatatcag taatataatt agcatataaa ataaaaaaaa
     4921 aaaatgctta tttgtcaccc gctgactgcg tttccttttg aattataaaa ggttccttat
     4981 ggttctggtc cactagcgtt ttattttctt ccttttcctg taatattttt aactttagct
     5041 catgtacttt gtgcttgtct atatgtttct cgtatttttc ctttgtcttg aaagtcctag
     5101 aacaattatt gtaaaagcat ctgtattttc ttccaacggt attcaataaa aggtttagcc
     5161 cattttcacc agactcaagt cgcttttcta aatccacttc attctgtaaa aactctacat
     5221 tattgttttc ggtaagtttt ctccttttcc tcaacctatt ggaaatcttc tcttcatcct
     5281 gctcattcga gtgtttgcta ttccccagtt gaactccgtg gtcttggaca agttgttgaa
     5341 tatcagaaat tttatatttt agttccagtg ggatatcttc ttccgtatga atagacccat
     5401 aatgcgtgag aagatcatgt tttctagaaa aagacatatc aggacatata tgacacttcc
     5461 aatttttggt tactagtgag tcgtcatgaa taatcatgtg catttgtaaa ccattttccc
     5521 ccacacatgg tttgctacaa ataggacatt ttaatttagg atgatcattt tttatatgcg
     5581 attgcaattg tgaccatatc cgaaactctt tgcaacaacc agcaaaagta cattggtaag
     5641 gattttctac ctcaggatcg tgatgtttag aaatgtgatt tcttagccta tatggcctct
     5701 gaaagcttat cgataccgtc gacctcgagg gggggcccgg tacccagctt ttgttccctt
     5761 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5821 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5881 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5941 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6001 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6061 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6121 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6181 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6241 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6301 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6361 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6421 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6481 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6541 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6601 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     6661 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6721 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6781 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6841 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6901 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6961 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7021 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7081 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7141 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7201 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7261 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7321 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7381 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7441 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7501 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7561 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7621 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7681 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7741 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7801 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7861 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7921 cgcacatttc cccgaaaagt gccac
//
