LOCUS       sequence-128287-        7534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of 2GFP tag at Atg11 C terminus, use auxotrophic marker 
            URA3(Candida albicans).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7534)
  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 7534)
  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: LD211
FEATURES             Location/Qualifiers
     source          1..7534
                     /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             1352..2065
                     /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             2102..2818
                     /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             2102..2815
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2839..3026
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2894..2918)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        3101..3444
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      4263..4460
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(5346..5366)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5346..5364)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5385..5401)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5385..5401)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5398..5420)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5409..5425
                     /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(5433..5439,5440..5457,5458..5463))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5478..5499
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5616..5633)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5787..6375)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5867..5886)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6546..7337,7338..7406))
                     /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     7169..7188
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7407..7511)
                     /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 ccgcggtgga tctgaaaagg atacacagca aagcattttt ctggggaata tcactcaatt
      721 atatgacaat aagctttatg aagttgccgt tatcaggaga ttcaaggata ttgagacctt
      781 ggccaaaaaa ttaactaaag aaaataagat caagagaacg ctattggaga gattccaaag
      841 ggaaaaggtg actttgagga actttcaaat tggcgattta gcattatttt tgccaacacg
      901 agaaaacgtt aattctgtcg ggtctatgtc gtcttccacc tcatcattga gctcatcatt
      961 ttcatctgtt gatttatcaa ctccaccgcc tttggatgct atgtctatac aatcgtctcc
     1021 ctcggtgata catagtaatg taatcaatca agccagtata tctggcaggg acaagaataa
     1081 actgatgagg ccttgggctg catttacagc ttttgaagaa agcacaagat attttttaaa
     1141 ggacgaaaaa ggattgacga aggggaaaga atggtttgta ggcagaattg tcactttgga
     1201 acatttcgta gcagatagcc cctctaataa tccatttaga ctaccaaagg gctcagtttg
     1261 gtttcaggtc actgcggttg tggtttcata ccagggagtt gcttcgtacg ctgcaggtcg
     1321 acggatcggt gacggtgctg gtttaattaa catgtctaaa ggtgaagaat tattcactgg
     1381 tgttgtccca attttggttg aattagatgg tgatgttaat ggtcacaaat tttctgtctc
     1441 cggtgaaggt gaaggtgatg ctacttacgg taaattgacc ttaaaattta tttgtactac
     1501 tggtaaattg ccagttccat ggccaacctt agtcactact ttaacttatg gtgttcaatg
     1561 tttttctaga tacccagatc atatgaaaca acatgacttt ttcaagtctg ccatgccaga
     1621 aggttatgtt caagaaagaa ctattttttt caaagatgac ggtaactaca agaccagagc
     1681 tgaagtcaag tttgaaggtg ataccttagt taatagaatc gaattaaaag gtattgattt
     1741 taaagaagat ggtaacattt taggtcacaa attggaatac aactataact ctcacaatgt
     1801 ttacatcatg gctgacaaac aaaagaatgg tatcaaagtt aacttcaaaa ttagacacaa
     1861 cattgaagat ggttctgttc aattagctga ccattatcaa caaaatactc caattggtga
     1921 tggtccagtc ttgttaccag acaaccatta cttatccact caatctgcct tatccaaaga
     1981 tccaaacgaa aagagagacc acatggtctt gttagaattt gttactgctg ctggtattac
     2041 ccatggtatt gatgaattgt acaaaattcc agttccaacg cgtgatctag agaagccatt
     2101 catgtctaaa ggtgaagaat tattcactgg tgttgtccca attttggttg aattagatgg
     2161 tgatgttaat ggtcacaaat tttctgtctc cggtgaaggt gaaggtgatg ctacttacgg
     2221 taaattgacc ttaaaattta tttgtactac tggtaaattg ccagttccat ggccaacctt
     2281 agtcactact ttaacttatg gtgttcaatg tttttctaga tacccagatc atatgaaaca
     2341 acatgacttt ttcaagtctg ccatgccaga aggttatgtt caagaaagaa ctattttttt
     2401 caaagatgac ggtaactaca agaccagagc tgaagtcaag tttgaaggtg ataccttagt
     2461 taatagaatc gaattaaaag gtattgattt taaagaagat ggtaacattt taggtcacaa
     2521 attggaatac aactataact ctcacaatgt ttacatcatg gctgacaaac aaaagaatgg
     2581 tatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggttctgttc aattagctga
     2641 ccattatcaa caaaatactc caattggtga tggtccagtc ttgttaccag acaaccatta
     2701 cttatccact caatctgcct tatccaaaga tccaaacgaa aagagagacc acatggtctt
     2761 gttagaattt gttactgctg ctggtattac ccatggtatg gatgaattat ataaataagg
     2821 cgcgccactt ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt
     2881 ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc
     2941 ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc
     3001 gctcttattg accacacctc taccggcaga tccgctaggg ataacagggt aatatagatc
     3061 tgtttagctt gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac
     3121 cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat
     3181 ttagcccata catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc
     3241 gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac
     3301 agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt
     3361 gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca
     3421 catcacatcc gaacataaac aaccatgaca gtcaacacta agacctatag tgagagagca
     3481 gaaactcatg cctcaccagt agcacagcga ttatttcgat taatggaact gaagaaaacc
     3541 aatttatgtg catcaattga cgttgatacc actaaggaat tccttgaatt aattgataaa
     3601 ttaggtcctt atgtatgctt aatcaagact catattgata taatcaatga tttttcctat
     3661 gaatccacta ttgaaccatt attagaactt tcacgtaaac atcaatttat gatttttgaa
     3721 gatagaaaat ttgctgatat tggtaatacc gtaaagaaac aatatattgg tggagtttat
     3781 aaaattagta gttgggcaga tattaccaat gctcatggtg tcactgggaa tggagtggtt
     3841 gaaggattaa aacagggagc taaagaaacc accaccaacc aagagccaag agggttattg
     3901 atgttagctg aattatcatc agtgggatca ttagcatatg gagaatattc tcaaaaaact
     3961 gttgaaattg ctaaatccga taaggaattt gttattggat ttattgccca acgtgatatg
     4021 ggtggccaag aagaaggatt tgattggctt attatgacac ctggagttgg attagatgat
     4081 aaaggtgatg gattaggaca acaatataga actgttgatg aagttgttag cactggaact
     4141 gatattatca ttgttggtag aggattgttt ggtaaaggaa gagatccaga tattgaaggt
     4201 aaaaggtata gaaatgctgg ttggaatgct tatttgaaaa agactggcca attataatca
     4261 gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata
     4321 ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca
     4381 tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt
     4441 gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca gtgtcgaaaa
     4501 cgagctcgaa ttcatcgatg atatcagatc cactagtggc ctatgctgaa cagtgctaaa
     4561 cgtttgtcac aaatgacaag attttaatta tgtatctatt taacattata atgcgataag
     4621 tcttgttatc gactaccaag tttcctggat atatctatca gatttttcca tttctttcag
     4681 gtattttttg gcagtttctt catcactaaa attgccccat ttttttaaca tttcaatgaa
     4741 tgttaatctg acttggatag gcattttacc tgatgagcca cataagaaaa aaacggcgtc
     4801 tttattaact accaaattag ttatttcctc acctaatctc cataagtagt cttggacata
     4861 tttaacacct ggtgaatttt cctcatctct ggaaaaggat gaatgtaagg caattttgcc
     4921 cttacggaac caatcttcca gcatatcttt ataaatataa tctttatctt tatatctaca
     4981 accgaagagg agtttgatat ctttagatat ctccgcttta accactgata ataatggtgc
     5041 tagaccgaca cccggtccga caagaatcat aggtttattc aaaaattcct ttttgattat
     5101 gtgattattt tgtaacttat accttatctg ttccccctct tgcaatcgtg caatgtagtt
     5161 agtgcaaatt cctcttcgaa ttttccttaa gatagtttta tattttacta tcgcaactgt
     5221 taattcaata ttaggatccc cagggccgct agatatagag tagtaccttg gtttaatgat
     5281 gggcaagtaa tctagcacat atttccaggg cactcgaggg ggggcccggt acccagcttt
     5341 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     5401 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5461 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5521 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5581 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5641 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5701 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5761 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5821 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5881 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5941 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6001 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6061 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6121 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6181 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     6241 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6301 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6361 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6421 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6481 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6541 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6601 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6661 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6721 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6781 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6841 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6901 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6961 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7021 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7081 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7141 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7201 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7261 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7321 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7381 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7441 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7501 ggggttccgc gcacatttcc ccgaaaagtg ccac
//
