LOCUS       sequence-128284-        8483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of 2GFP tag at Atg6 C terminus, use auxotrophic marker 
            URA3(Candida albicans).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8483)
  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 8483)
  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: LD206
FEATURES             Location/Qualifiers
     source          1..8483
                     /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"
     primer_bind     677..693
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     677..693
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             1417..2130
                     /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             2167..2883
                     /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             2167..2880
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2904..3091
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2959..2983)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        3166..3509
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4328..4525
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(6249..6265)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6250..6266)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(6295..6315)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6295..6313)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6334..6350)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6334..6350)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6347..6369)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6358..6374
                     /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(6382..6388,6389..6406,6407..6412))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6427..6448
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6565..6582)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6736..7324)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6816..6835)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7495..8286,8287..8355))
                     /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     8118..8137
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8356..8460)
                     /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 ccgcggtggc ggccgctcta gaactagtgg atcccctttg ccactataaa tggattgaga
      721 ttaggcagta taccagaatc agtggtacct tggaaggaaa tcaatgcagc actgggccaa
      781 ttgattcttt tactggctac cataaacaaa aacctgaaga taaatttagt agactacgaa
      841 ttacaaccga tgggttcatt ttccaaaatc aagaaaagaa tggttaacag tgtcgaatat
      901 aataattcca ctacgaacgc tcccggtgat tggttgattc tgcctgtcta ctatgatgag
      961 aatttcaatt tggggagaat cttccgcaag gaaacgaagt ttgataaatc attagaaaca
     1021 acattagaaa taataagcga aatcactcgg cagctttcta cgatagcgtc atcatattca
     1081 tctcaaacac tcacaacaag ccaagatgaa tcttcaatga ataacgcaaa tgacgttgaa
     1141 aatagtactt ccattctcga gctaccctat atcatgaata aggacaagat taatggatta
     1201 tccgtcaaat tgcatggtag cagtcccaac cttgaatgga caacggctat gaaattttta
     1261 ttaaccaatg taaagtggtt gttagcgttt tcttccaatt tactatccaa gtcaattacc
     1321 ttaagcccta cagtgaatta caatgataag accatcagcg gaaacgcttc gtacgctgca
     1381 ggtcgacgga tcggtgacgg tgctggttta attaacatgt ctaaaggtga agaattattc
     1441 actggtgttg tcccaatttt ggttgaatta gatggtgatg ttaatggtca caaattttct
     1501 gtctccggtg aaggtgaagg tgatgctact tacggtaaat tgaccttaaa atttatttgt
     1561 actactggta aattgccagt tccatggcca accttagtca ctactttaac ttatggtgtt
     1621 caatgttttt ctagataccc agatcatatg aaacaacatg actttttcaa gtctgccatg
     1681 ccagaaggtt atgttcaaga aagaactatt tttttcaaag atgacggtaa ctacaagacc
     1741 agagctgaag tcaagtttga aggtgatacc ttagttaata gaatcgaatt aaaaggtatt
     1801 gattttaaag aagatggtaa cattttaggt cacaaattgg aatacaacta taactctcac
     1861 aatgtttaca tcatggctga caaacaaaag aatggtatca aagttaactt caaaattaga
     1921 cacaacattg aagatggttc tgttcaatta gctgaccatt atcaacaaaa tactccaatt
     1981 ggtgatggtc cagtcttgtt accagacaac cattacttat ccactcaatc tgccttatcc
     2041 aaagatccaa acgaaaagag agaccacatg gtcttgttag aatttgttac tgctgctggt
     2101 attacccatg gtattgatga attgtacaaa attccagttc caacgcgtga tctagagaag
     2161 ccattcatgt ctaaaggtga agaattattc actggtgttg tcccaatttt ggttgaatta
     2221 gatggtgatg ttaatggtca caaattttct gtctccggtg aaggtgaagg tgatgctact
     2281 tacggtaaat tgaccttaaa atttatttgt actactggta aattgccagt tccatggcca
     2341 accttagtca ctactttaac ttatggtgtt caatgttttt ctagataccc agatcatatg
     2401 aaacaacatg actttttcaa gtctgccatg ccagaaggtt atgttcaaga aagaactatt
     2461 tttttcaaag atgacggtaa ctacaagacc agagctgaag tcaagtttga aggtgatacc
     2521 ttagttaata gaatcgaatt aaaaggtatt gattttaaag aagatggtaa cattttaggt
     2581 cacaaattgg aatacaacta taactctcac aatgtttaca tcatggctga caaacaaaag
     2641 aatggtatca aagttaactt caaaattaga cacaacattg aagatggttc tgttcaatta
     2701 gctgaccatt atcaacaaaa tactccaatt ggtgatggtc cagtcttgtt accagacaac
     2761 cattacttat ccactcaatc tgccttatcc aaagatccaa acgaaaagag agaccacatg
     2821 gtcttgttag aatttgttac tgctgctggt attacccatg gtatggatga attatataaa
     2881 taaggcgcgc cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat
     2941 aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa
     3001 aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagtatg
     3061 aggtcgctct tattgaccac acctctaccg gcagatccgc tagggataac agggtaatat
     3121 agatctgttt agcttgcctc gtccccgccg ggtcacccgg ccagcgacat ggaggcccag
     3181 aataccctcc ttgacagtct tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg
     3241 tacatttagc ccatacatcc ccatgtataa tcatttgcat ccatacattt tgatggccgc
     3301 acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct gcgagcaggg aaacgctccc
     3361 ctcacagacg cgttgaattg tccccacgcc gcgcccctgt agagaaatat aaaaggttag
     3421 gatttgccac tgaggttctt ctttcatata cttcctttta aaatcttgct aggatacagt
     3481 tctcacatca catccgaaca taaacaacca tgacagtcaa cactaagacc tatagtgaga
     3541 gagcagaaac tcatgcctca ccagtagcac agcgattatt tcgattaatg gaactgaaga
     3601 aaaccaattt atgtgcatca attgacgttg ataccactaa ggaattcctt gaattaattg
     3661 ataaattagg tccttatgta tgcttaatca agactcatat tgatataatc aatgattttt
     3721 cctatgaatc cactattgaa ccattattag aactttcacg taaacatcaa tttatgattt
     3781 ttgaagatag aaaatttgct gatattggta ataccgtaaa gaaacaatat attggtggag
     3841 tttataaaat tagtagttgg gcagatatta ccaatgctca tggtgtcact gggaatggag
     3901 tggttgaagg attaaaacag ggagctaaag aaaccaccac caaccaagag ccaagagggt
     3961 tattgatgtt agctgaatta tcatcagtgg gatcattagc atatggagaa tattctcaaa
     4021 aaactgttga aattgctaaa tccgataagg aatttgttat tggatttatt gcccaacgtg
     4081 atatgggtgg ccaagaagaa ggatttgatt ggcttattat gacacctgga gttggattag
     4141 atgataaagg tgatggatta ggacaacaat atagaactgt tgatgaagtt gttagcactg
     4201 gaactgatat tatcattgtt ggtagaggat tgtttggtaa aggaagagat ccagatattg
     4261 aaggtaaaag gtatagaaat gctggttgga atgcttattt gaaaaagact ggccaattat
     4321 aatcagtact gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt
     4381 ttatattgta gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct
     4441 cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg
     4501 tatgtgaatg ctggtcgcta tactgctgtc gattcgatac taacgccgcc atccagtgtc
     4561 gaaaacgagc tcgaattcat cgatgatatc agatccacta gtggcctatg ctaagtagtt
     4621 catataaatg cattttttca taaatgtgat aaagggaaaa tttctttttc cacttgaaag
     4681 atatgtacgt atttaagaac tgatcttctt gaccactttt tttatgcgtg caactttaaa
     4741 tgctttcata catttccttg gtctttcatc agatgatatg ccagatatca acaattatag
     4801 acctaatata acattgagtt attttttaca cgtataataa taactccttc tccggcaatg
     4861 aatattttgc ttccaaattg caaaattcaa gatttaatta taaaaactct gaagtaatag
     4921 accacctatt tcaaaaatat tccgtatttt tccctctcta taaaaacaaa gggcttccaa
     4981 aatatccatt ccaataaacg gtaaactcac ccatttatga ttgggacaag agcaaaaaaa
     5041 aaggaaattg ggaaaaaaaa agagaaagaa agcaacagaa taaagggaac ttgataaaac
     5101 tgaagtaccg tgggtcaagt agcgtgaaag agtcaattca tcgactaatg gatgcataaa
     5161 acgtatgact gtatagtact ggagtgaaaa cgaagagata aagaaaggaa aaagggagat
     5221 taataaaaag aaaaaaatta aaggagttct atatttggat tagtctttta ttctttctgc
     5281 ttaccaccat gaagcgttgc tatttccgta accgtttttc tcattatcat ttctaaaccc
     5341 acctccaatc gaaccccaat ttgatgttcc agtattcctt ggcctcgtgc tcccaaatga
     5401 gccatttcca ccatgtttac ggtagtctct tgaaccgtta ttacgactat tgaaaaaacc
     5461 tcctcctccc ctagttcttc cacctcttga attctgtctt gacaggtcac ttaaaaacgt
     5521 gggaacttct tgattcgctt catttaaaat ttccatcaaa cccttcacaa tattctgatt
     5581 attactattg aagaatgaag tagccacgcc tgtgttacca gcacggcctg ttctaccaat
     5641 tctgtgaacg tagtcatcga tgtcggaggg caaatcataa ttaatgacat gtgtgacatt
     5701 cggtatatcc aaacctctcg ctgctacagc tgttgcgacc aggatatcag ctacgttagc
     5761 tttgaaagca gataaggcac gttcacgttc agcctgtgtg cggtcaccat gtatggctgt
     5821 agctttgaaa ttttgcatga tcaaaaaatc tgtgagttga tccgccattc ttttcgtttc
     5881 aacaaaaatc aatgtcaatc ccttatgctc tgcagacagt aaatctaaca acgcagactt
     5941 tttgtccatg tcatcgacat ataatattct ttgcgtgata ttttctgagg tggaaccgac
     6001 tcttcctact gataaaaaaa tataattatc taagaaatcg cgtgccaaat gctggatatc
     6061 tactgggaat gtggcggaaa acatcaaggt ctgtctattt tccacagagg gcatatcaca
     6121 ttcttcaact atatggcgaa tttggggttc gaatcccata tctagcattc tatcagcctc
     6181 atccaatacc aggtatttga tattagccaa agaaactttt ccgcgttcta ataagtcatt
     6241 aagcttatcg ataccgtcga cctcgagggg gggcccggta cccagctttt gttcccttta
     6301 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6361 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6421 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6481 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6541 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6601 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6661 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6721 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6781 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6841 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6901 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6961 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     7021 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7081 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7141 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     7201 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7261 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7321 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7381 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7441 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7501 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7561 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7621 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7681 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7741 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7801 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7861 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7921 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7981 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8041 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8101 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8161 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8221 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8281 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8341 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8401 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8461 cacatttccc cgaaaagtgc cac
//
