LOCUS       sequence-128244-        7195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of GFP 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 7195)
  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 7195)
  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: LD113
FEATURES             Location/Qualifiers
     source          1..7195
                     /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..2148
                     /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      2169..2356
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2224..2248)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        2431..2774
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3593..3790
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(4961..4977)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4962..4978)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5007..5027)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5007..5025)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5046..5062)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5046..5062)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5059..5081)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5070..5086
                     /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(5094..5100,5101..5118,5119..5124))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5139..5160
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5277..5294)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5448..6036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5528..5547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6207..6998,6999..7067))
                     /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     6830..6849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7068..7172)
                     /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 acaaataagg cgcgccactt
     2161 ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa
     2221 taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga
     2281 gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc gctcttattg
     2341 accacacctc taccggcaga tccgctaggg ataacagggt aatatagatc tgtttagctt
     2401 gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac
     2461 agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata
     2521 catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa
     2581 aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg
     2641 aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg
     2701 ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc
     2761 gaacataaac aaccatgaca gtcaacacta agacctatag tgagagagca gaaactcatg
     2821 cctcaccagt agcacagcga ttatttcgat taatggaact gaagaaaacc aatttatgtg
     2881 catcaattga cgttgatacc actaaggaat tccttgaatt aattgataaa ttaggtcctt
     2941 atgtatgctt aatcaagact catattgata taatcaatga tttttcctat gaatccacta
     3001 ttgaaccatt attagaactt tcacgtaaac atcaatttat gatttttgaa gatagaaaat
     3061 ttgctgatat tggtaatacc gtaaagaaac aatatattgg tggagtttat aaaattagta
     3121 gttgggcaga tattaccaat gctcatggtg tcactgggaa tggagtggtt gaaggattaa
     3181 aacagggagc taaagaaacc accaccaacc aagagccaag agggttattg atgttagctg
     3241 aattatcatc agtgggatca ttagcatatg gagaatattc tcaaaaaact gttgaaattg
     3301 ctaaatccga taaggaattt gttattggat ttattgccca acgtgatatg ggtggccaag
     3361 aagaaggatt tgattggctt attatgacac ctggagttgg attagatgat aaaggtgatg
     3421 gattaggaca acaatataga actgttgatg aagttgttag cactggaact gatattatca
     3481 ttgttggtag aggattgttt ggtaaaggaa gagatccaga tattgaaggt aaaaggtata
     3541 gaaatgctgg ttggaatgct tatttgaaaa agactggcca attataatca gtactgacaa
     3601 taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt
     3661 tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc
     3721 agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt
     3781 cgctatactg ctgtcgattc gatactaacg ccgccatcca gtgtcgaaaa cgagctcgaa
     3841 ttcatcgatg atatcagatc cactagtggc ctatgctaaa aatggtaaag ttttatttta
     3901 aagggcacaa ctaaagaaaa ataatcaaaa aaaaggtgaa atagatatgc aagaaaagaa
     3961 aaaaaaatac aatactagta aactgaaata aaaaagatta gccataatgg atttgtttct
     4021 ctttgttatc gatattctta cctcttgaag atccatgtgt ctatcatttg aattgaatat
     4081 ataattatat acagtgagat ataccaattg cagtaacaaa atggcaaata tgtatatcag
     4141 taatataatt agcatataaa ataaaaaaaa aaaatgctta tttgtcaccc gctgactgcg
     4201 tttccttttg aattataaaa ggttccttat ggttctggtc cactagcgtt ttattttctt
     4261 ccttttcctg taatattttt aactttagct catgtacttt gtgcttgtct atatgtttct
     4321 cgtatttttc ctttgtcttg aaagtcctag aacaattatt gtaaaagcat ctgtattttc
     4381 ttccaacggt attcaataaa aggtttagcc cattttcacc agactcaagt cgcttttcta
     4441 aatccacttc attctgtaaa aactctacat tattgttttc ggtaagtttt ctccttttcc
     4501 tcaacctatt ggaaatcttc tcttcatcct gctcattcga gtgtttgcta ttccccagtt
     4561 gaactccgtg gtcttggaca agttgttgaa tatcagaaat tttatatttt agttccagtg
     4621 ggatatcttc ttccgtatga atagacccat aatgcgtgag aagatcatgt tttctagaaa
     4681 aagacatatc aggacatata tgacacttcc aatttttggt tactagtgag tcgtcatgaa
     4741 taatcatgtg catttgtaaa ccattttccc ccacacatgg tttgctacaa ataggacatt
     4801 ttaatttagg atgatcattt tttatatgcg attgcaattg tgaccatatc cgaaactctt
     4861 tgcaacaacc agcaaaagta cattggtaag gattttctac ctcaggatcg tgatgtttag
     4921 aaatgtgatt tcttagccta tatggcctct gaaagcttat cgataccgtc gacctcgagg
     4981 gggggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     5041 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5101 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5161 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5221 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5281 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5341 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5401 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5461 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5521 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5581 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5641 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5701 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5761 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5821 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5881 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5941 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6001 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6061 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6121 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6181 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6241 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6301 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6361 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6421 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6481 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6541 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6601 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6661 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6721 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6781 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6841 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6901 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6961 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7021 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7081 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7141 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccac
//
