LOCUS       sequence-121962-        8034 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Integration of GFP tag at Atg24 C terminus, use auxotrophic marker 
            URA3(Candida albicans).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8034)
  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 8034)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: LD124
FEATURES             Location/Qualifiers
     source          1..8034
                     /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             2487..3203
                     /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             2487..3200
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      3224..3411
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3279..3303)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        3486..3829
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4648..4845
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(5846..5866)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5846..5864)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5885..5901)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5885..5901)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5898..5920)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5909..5925
                     /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(5933..5939,5940..5957,5958..5963))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5978..5999
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6116..6133)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6287..6875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6367..6386)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7046..7837,7838..7906))
                     /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     7669..7688
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7907..8011)
                     /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 ccgcggtttc tgctgaaagc ccaagctttc ataaattaat aggtttcgtg gtctagtcgg
      721 ttatggcatc tgcttaacac gcagaacgtc cccagttcga tcctgggcga aatcaatttt
      781 ttgttttttt aattcacaag aataacctgg tattgttata agcaagccac cctttgcgta
      841 cgatcaaaca ccctataaaa tctatatgca atactgagat attgtatttt ttctccttta
      901 attgtttata gttcagaata acagttcaaa aatctatgcc gcgctcacag ggtcctgccg
      961 ggtattcgcc ctccagcttg aaaggggtgg caatgcccaa gagcgatgta tagtttagta
     1021 tcaaggaatt gagagaggta ttacaccaag gatttccgtg aacgaagtga cgacgcaaga
     1081 ttaacagatt aaattctggt agtaggctaa gccgttatta aactttattt acggtatacc
     1141 acaatactgc tctttttgtt gaggatatga cagacaaagg caagaacgat ttaacaagca
     1201 aagctaagga caaagccaga ggcaaccccg agaagccacc atattggttt gaaatcattg
     1261 tttctgatcc acaaaaaaga actggtgacc caggctcttc atctggctac gtgagttatc
     1321 agatatcaac aaagactaac aacacgtcat tttatgataa tcgtggcgat ccagaatcta
     1381 ttatagttgt acatagaaga tacagtgacc tattgctcct gcatgatatt ttactcaaca
     1441 ggttccccac ctgcatcatc ccgccattac ctgataagaa agtttttcaa tacatagcag
     1501 gcgataggtt tagccaaaga tttacacaaa agagatgtca tagtttgcaa aatttcctaa
     1561 gaagggtctc tttgcatccc gatctttctc aatcaaaagt cttcaaaact tttttggtga
     1621 gtaaagattg ggaatctcac cgcaaagtcc tccaggatag cttgcaacca aataaagatg
     1681 aagttactga tgcttttatg aatgccttca aaacagttca caagcaaaac gaagaattca
     1741 ctgagataag ggagaaaagt gataaactag atcggactgt gacaaagata gataaacttt
     1801 ttcataaggt tgtgaaaaaa aatgattcga tgtctgaaga ctatacaaag ttgggcagta
     1861 atttacagga attacaagaa ttggtcacag gcgaaaacga ggaacttgct gcgaagttga
     1921 agatttttaa tgaaggtgta acacagctgt catatggctt gcaagatctt acaaagtatc
     1981 tggattacga atatattgtt gatttgaagg atcttgaaca ctacattgat agtatgcgac
     2041 agctgatcaa attgaaagac cagaaacaga tagactacga agaactaagt gattatttaa
     2101 ccaggtcaat caaagaaaaa aataacttga tttctggata cggaggtagt aatttctttg
     2161 ccaataaatt ggaagaacta gcgggaataa atcaggaggc ttcacgtaga gaaaaaatca
     2221 ataaactaga aggcaaaata acgtcattaa caggtgaatt agaaaacgct aaaaaggtgg
     2281 cggatggatt cgaacaggaa tgcttgaagg aaatagacca tttcgaaagt gtaaaaacgg
     2341 ctgaaatcaa gaaatctctg ggctcactag cggaccatca tatcgaattc tacgaaagga
     2401 tacttgaagc atgggaaaaa gtagacgata gtctagcttc gtacgctgca ggtcgacgga
     2461 tcggtgacgg tgctggttta attaacatgt ctaaaggtga agaattattc actggtgttg
     2521 tcccaatttt ggttgaatta gatggtgatg ttaatggtca caaattttct gtctccggtg
     2581 aaggtgaagg tgatgctact tacggtaaat tgaccttaaa atttatttgt actactggta
     2641 aattgccagt tccatggcca accttagtca ctactttaac ttatggtgtt caatgttttt
     2701 ctagataccc agatcatatg aaacaacatg actttttcaa gtctgccatg ccagaaggtt
     2761 atgttcaaga aagaactatt tttttcaaag atgacggtaa ctacaagacc agagctgaag
     2821 tcaagtttga aggtgatacc ttagttaata gaatcgaatt aaaaggtatt gattttaaag
     2881 aagatggtaa cattttaggt cacaaattgg aatacaacta taactctcac aatgtttaca
     2941 tcatggctga caaacaaaag aatggtatca aagttaactt caaaattaga cacaacattg
     3001 aagatggttc tgttcaatta gctgaccatt atcaacaaaa tactccaatt ggtgatggtc
     3061 cagtcttgtt accagacaac cattacttat ccactcaatc tgccttatcc aaagatccaa
     3121 acgaaaagag agaccacatg gtcttgttag aatttgttac tgctgctggt attacccatg
     3181 gtatggatga attgtacaaa taaggcgcgc cacttctaaa taagcgaatt tcttatgatt
     3241 tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga
     3301 ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt
     3361 gctttctcag gtatagtatg aggtcgctct tattgaccac acctctaccg gcagatccgc
     3421 tagggataac agggtaatat agatctgttt agcttgcctc gtccccgccg ggtcacccgg
     3481 ccagcgacat ggaggcccag aataccctcc ttgacagtct tgacgtgcgc agctcagggg
     3541 catgatgtga ctgtcgcccg tacatttagc ccatacatcc ccatgtataa tcatttgcat
     3601 ccatacattt tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct
     3661 gcgagcaggg aaacgctccc ctcacagacg cgttgaattg tccccacgcc gcgcccctgt
     3721 agagaaatat aaaaggttag gatttgccac tgaggttctt ctttcatata cttcctttta
     3781 aaatcttgct aggatacagt tctcacatca catccgaaca taaacaacca tgacagtcaa
     3841 cactaagacc tatagtgaga gagcagaaac tcatgcctca ccagtagcac agcgattatt
     3901 tcgattaatg gaactgaaga aaaccaattt atgtgcatca attgacgttg ataccactaa
     3961 ggaattcctt gaattaattg ataaattagg tccttatgta tgcttaatca agactcatat
     4021 tgatataatc aatgattttt cctatgaatc cactattgaa ccattattag aactttcacg
     4081 taaacatcaa tttatgattt ttgaagatag aaaatttgct gatattggta ataccgtaaa
     4141 gaaacaatat attggtggag tttataaaat tagtagttgg gcagatatta ccaatgctca
     4201 tggtgtcact gggaatggag tggttgaagg attaaaacag ggagctaaag aaaccaccac
     4261 caaccaagag ccaagagggt tattgatgtt agctgaatta tcatcagtgg gatcattagc
     4321 atatggagaa tattctcaaa aaactgttga aattgctaaa tccgataagg aatttgttat
     4381 tggatttatt gcccaacgtg atatgggtgg ccaagaagaa ggatttgatt ggcttattat
     4441 gacacctgga gttggattag atgataaagg tgatggatta ggacaacaat atagaactgt
     4501 tgatgaagtt gttagcactg gaactgatat tatcattgtt ggtagaggat tgtttggtaa
     4561 aggaagagat ccagatattg aaggtaaaag gtatagaaat gctggttgga atgcttattt
     4621 gaaaaagact ggccaattat aatcagtact gacaataaaa agattcttgt tttcaagaac
     4681 ttgtcatttg tatagttttt ttatattgta gttgttctat tttaatcaaa tgttagcgtg
     4741 atttatattt tttttcgcct cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa
     4801 gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc gattcgatac
     4861 taacgccgcc atccagtgtc gaaaacgagc tcgaattcat cgatgatatc agatccacta
     4921 gtggcctatg ctagtggtgc tcttaatgtt ttcccattac tactgataat accttgggca
     4981 ctacgattat ctgatcacat ttaacagaga aatttaaaaa aagaaaaata tctatataaa
     5041 aatatgtaaa tgttgctatt attttatgat gttctgattt ttttcgtcaa accttagatt
     5101 tcttatgtac attaaactta aacattttgc attgttcttc cagttcttgg attatattat
     5161 cttggtggga gtcaatcaaa tcccaatcga cgctattact ggacaagtca actttattcg
     5221 caaaacaagt cctgtaatca tcaccaaaag tctcaatgaa tgcttcttca ttaagatact
     5281 ttgaccattc cataggcgga aacgtgtttt tatccaacac tctgtcactc ttggaccgtg
     5341 gctttggcgc cctttcattt tgtcttacat aaaagtatct cagcagcatt attgcttctt
     5401 tcctcaagat ccccggtata ctctcgtacc ctgccgcact attgttcttg ggcactaatg
     5461 tacacgtatc atgatttact gacaagacag taccgttgcc tccaaatctc tcgttgccac
     5521 aaccgaacac cacctttcca atgtctaatt gcttgagagc agatgcacac attatacacg
     5581 gttctacagt aacatataga gtaatgtctt tgaacacgtc aacaactcct cgggagccca
     5641 acatcgcctt gatctgatcg atccccatga actcagcatg tgctactcca gttaatgact
     5701 tgttcgtatc gttcatgcca taagccatca cctggccagt tggcgtatgt acaaagatgc
     5761 aagctaccgg tgtctcatcg tggtcaagag cgtatctagc caaacggaca gcctcgaggg
     5821 ggggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     5881 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5941 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     6001 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6061 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6121 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6181 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6241 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6301 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6361 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6421 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6481 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6541 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6601 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6661 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6721 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6781 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6841 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6901 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6961 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7021 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7081 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7141 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7201 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7261 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7321 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7381 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7441 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7501 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7561 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7621 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7681 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7741 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7801 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7861 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7921 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7981 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccac
//
