LOCUS       sequence-128242-        7733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of GFP 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 7733)
  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 7733)
  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: LD106
FEATURES             Location/Qualifiers
     source          1..7733
                     /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..2133
                     /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      2154..2341
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2209..2233)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2416..2759
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3578..3775
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(5499..5515)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5500..5516)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5545..5565)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5545..5563)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5584..5600)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5584..5600)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5597..5619)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5608..5624
                     /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(5632..5638,5639..5656,5657..5662))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5677..5698
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5815..5832)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5986..6574)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6066..6085)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6745..7536,7537..7605))
                     /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     7368..7387
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7606..7710)
                     /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 taaggcgcgc cacttctaaa taagcgaatt
     2161 tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat
     2221 tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt
     2281 aggtcaggtt gctttctcag gtatagtatg aggtcgctct tattgaccac acctctaccg
     2341 gcagatccgc tagggataac agggtaatat agatctgttt agcttgcctc gtccccgccg
     2401 ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct tgacgtgcgc
     2461 agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc ccatgtataa
     2521 tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg
     2581 ctgcagacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg tccccacgcc
     2641 gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt ctttcatata
     2701 cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca taaacaacca
     2761 tgacagtcaa cactaagacc tatagtgaga gagcagaaac tcatgcctca ccagtagcac
     2821 aacgattatt tcgattaatg gaactgaaga aaaccaattt atgtgcatca attgatgttg
     2881 ataccactaa ggaattcctt gaattaattg ataaattggg tccttatgta tgcttaatca
     2941 agactcatat tgatataatc aatgattttt cctatgaatc cactattgaa ccattattag
     3001 aactttcacg taaacatcaa tttatgattt ttgaagatag aaaatttgct gatattggta
     3061 ataccgtgaa gaaacaatat attggtggag tttataaaat tagtagttgg gcagatatta
     3121 ctaatgctca tggtgtcact gggaatggag tagttgaagg attaaaacag ggagctaaag
     3181 aaaccaccac caaccaagag ccaagagggt tattgatgtt agctgaatta tcatcagtgg
     3241 gatcattagc atatggagaa tattctcaaa aaactgttga aattgctaaa tccgataagg
     3301 aatttgttat tggatttatt gcccaacgtg atatgggtgg acaagaagaa ggatttgatt
     3361 ggcttattat gacacctgga gttggattag atgataaagg tgatggatta ggacaacaat
     3421 atagaactgt tgatgaagtt gttagcactg gaactgatat tatcattgtt ggtagaggat
     3481 tgtttggtaa aggaagagat ccagatattg aaggtaaaag gtatagagat gctggttgga
     3541 atgcttattt gaaaaagact ggccaattat aatcagtact gacaataaaa agattcttgt
     3601 tttcaagaac ttgtcatttg tatagttttt ttatattgta gttgttctat tttaatcaaa
     3661 tgttagcgtg atttatattt tttttcgcct cgacatcatc tgcccagatg cgaagttaag
     3721 tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc
     3781 gattcgatac taacgccgcc atccagtgtc gaaaacgagc tcgaattcat cgatgatatc
     3841 agatccacta gtggcctatg ctaagtagtt catataaatg cattttttca taaatgtgat
     3901 aaagggaaaa tttctttttc cacttgaaag atatgtacgt atttaagaac tgatcttctt
     3961 gaccactttt tttatgcgtg caactttaaa tgctttcata catttccttg gtctttcatc
     4021 agatgatatg ccagatatca acaattatag acctaatata acattgagtt attttttaca
     4081 cgtataataa taactccttc tccggcaatg aatattttgc ttccaaattg caaaattcaa
     4141 gatttaatta taaaaactct gaagtaatag accacctatt tcaaaaatat tccgtatttt
     4201 tccctctcta taaaaacaaa gggcttccaa aatatccatt ccaataaacg gtaaactcac
     4261 ccatttatga ttgggacaag agcaaaaaaa aaggaaattg ggaaaaaaaa agagaaagaa
     4321 agcaacagaa taaagggaac ttgataaaac tgaagtaccg tgggtcaagt agcgtgaaag
     4381 agtcaattca tcgactaatg gatgcataaa acgtatgact gtatagtact ggagtgaaaa
     4441 cgaagagata aagaaaggaa aaagggagat taataaaaag aaaaaaatta aaggagttct
     4501 atatttggat tagtctttta ttctttctgc ttaccaccat gaagcgttgc tatttccgta
     4561 accgtttttc tcattatcat ttctaaaccc acctccaatc gaaccccaat ttgatgttcc
     4621 agtattcctt ggcctcgtgc tcccaaatga gccatttcca ccatgtttac ggtagtctct
     4681 tgaaccgtta ttacgactat tgaaaaaacc tcctcctccc ctagttcttc cacctcttga
     4741 attctgtctt gacaggtcac ttaaaaacgt gggaacttct tgattcgctt catttaaaat
     4801 ttccatcaaa cccttcacaa tattctgatt attactattg aagaatgaag tagccacgcc
     4861 tgtgttacca gcacggcctg ttctaccaat tctgtgaacg tagtcatcga tgtcggaggg
     4921 caaatcataa ttaatgacat gtgtgacatt cggtatatcc aaacctctcg ctgctacagc
     4981 tgttgcgacc aggatatcag ctacgttagc tttgaaagca gataaggcac gttcacgttc
     5041 agcctgtgtg cggtcaccat gtatggctgt agctttgaaa ttttgcatga tcaaaaaatc
     5101 tgtgagttga tccgccattc ttttcgtttc aacaaaaatc aatgtcaatc ccttatgctc
     5161 tgcagacagt aaatctaaca acgcagactt tttgtccatg tcatcgacat ataatattct
     5221 ttgcgtgata ttttctgagg tggaaccgac tcttcctact gataaaaaaa tataattatc
     5281 taagaaatcg cgtgccaaat gctggatatc tactgggaat gtggcggaaa acatcaaggt
     5341 ctgtctattt tccacagagg gcatatcaca ttcttcaact atatggcgaa tttggggttc
     5401 gaatcccata tctagcattc tatcagcctc atccaatacc aggtatttga tattagccaa
     5461 agaaactttt ccgcgttcta ataagtcatt aagcttatcg ataccgtcga cctcgagggg
     5521 gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     5581 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5641 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5701 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5761 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5821 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5881 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5941 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6001 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6061 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6121 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6181 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6241 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6301 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6361 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6421 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6481 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6541 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6601 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6661 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6721 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6781 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6841 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6901 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6961 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7021 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7081 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7141 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7201 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7261 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7321 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7381 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7441 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7501 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7561 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7621 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7681 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac
//
