LOCUS       sequence-272372-        7801 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of GFP-PEP12 as an additional copy in genome, use 
            auxotrophic marker URA3(Kluyveromyces lactis).Marker for yeast late 
            endosome..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7801)
  AUTHORS   Zhu J, Zhang ZT, Tang SW, Zhao BS, Li H, Song JZ, Li D, Xie Z
  TITLE     A Validated Set of Fluorescent-Protein-Based Markers for Major 
            Organelles in Yeast (Saccharomyces cerevisiae).
  JOURNAL   MBio. 2019 Sep 3;10(5). pii: mBio.01691-19. doi: 
            10.1128/mBio.01691-19.
  PUBMED    31481383
REFERENCE   2  (bases 1 to 7801)
  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: "MBio."; 
            date: "2019-09-3"; volume: "10(5). pii: mBio.01691-19. doi"; pages: 
            " 10.1128/mBio.01691-19"
COMMENT     Sequence Label: ZJOM157
FEATURES             Location/Qualifiers
     source          1..7801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..217
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     protein_bind    complement(1161..1194)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             2264..2977
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2264..2977
                     /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"
     promoter        complement(4562..4580)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4601..4617)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4601..4617)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4614..4636)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4625..4641
                     /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(4649..4655,4656..4673,4674..4679))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4694..4715
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4832..4849)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5003..5591)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5083..5102)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5762..6553,6554..6622))
                     /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     6385..6404
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6623..6727)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6753..7208)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6890..6911)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7102..7121
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7335..7357
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7349..7366
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7350..7366
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7373..7392
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7373..7391
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ttcaattcat cttttttttt tttgttcttt tttttgattc cggtttcttt gaaatttttt
       61 tgattcggta atctccgagc agaaggaaga acgaaggaag gagcacagac ttagattggt
      121 atatatacgc atatgtggtg ttgaagaaac atgaaattgc ccagtattct taacccaact
      181 gcacagaaca aaaacctgca ggaaacgaag ataaatcctc gagatgtcca caaaatcata
      241 taccagtaga gctgagactc atgcaagtcc ggttgcatcg aaacttttac gtttaatgga
      301 tgaaaagaag accaatttgt gtgcttctct tgacgttcgt tcgactgatg agctattgaa
      361 acttgttgaa acgttgggtc catacatttg ccttttgaaa acacacgttg atatcttgga
      421 tgatttcagt tatgagggta ctgtcgttcc attgaaagca ttggcagaga aatacaagtt
      481 cttgatattt gaggacagaa aattcgccga tatcggtaac acagtcaaat tacaatatac
      541 atcgggcgtt taccgtatcg cagaatggtc tgatatcacc aacgcccacg gggttactgg
      601 tgctggtatt gttgctggct tgaaacaagg tgcgcaagag gtcaccaaag aaccaagggg
      661 attattgatg cttgctgaat tgtcttccaa gggttctcta gcacacggtg aatatactaa
      721 gggtaccgtt gatattgcaa agagtgataa agatttcgtt attgggttca ttgctcagaa
      781 cgatatggga ggaagagaag aagggtttga ttggctaatc atgaccccag gtgtaggttt
      841 agacgacaaa ggcgatgcat tgggtcagca gtacagaacc gtcgacgaag ttgtaagtgg
      901 tggatcagat atcatcattg ttggcagagg acttttcgcc aagggtagag atcctaaggt
      961 tgaaggtgaa agatacagaa atgctggatg ggaagcgtac caaaagagaa tcagcgctcc
     1021 ccattaatta tacaggaaac ttaatagaac aaatcacata tttaatctaa tagccacctg
     1081 cattggcacg gtgcaacact acttcaactt catcttacaa aaagatcacg tgatctgttg
     1141 tattgggatc tctagaccta ataacttcgt atagcataca ttatacgaag ttatattaag
     1201 ggttgtcgac ctgcagcgta catcgataag cttgatatcg aattcctgca gcccggggga
     1261 tcctacttag tcctttggtc aaaaatattt cagaacaaag tgttgtcaat gaaggatata
     1321 attctgatag ggttttgact atgatggaca atggatcatt tccaacgtct atttctgcct
     1381 ccacaattaa gctactcatg aaatcgacca ctgctttatc aaagtctttt tggaagcttg
     1441 attgtaattc agcaaaaatt ataagcatca tcgattttac ttcatcttcg cttatgcgca
     1501 aaataagttc cttcactaaa aatcgcacct ccttgaaatt gtacttgaac ttattcaata
     1561 gttgtagaat tatgctcagt aaatacgtga cgtgggtgtc ctcgccatag tgtatcctaa
     1621 tttgcagctc atttaacaat gcaattgttg cgtctttatt ggaaaagcaa ccaacgaaaa
     1681 cgtggatgga tgatcttgat aatatttcaa ataaattgag tgcgaaggga atatcattct
     1741 gtataagttt agataaatgt acgcgggact cagaatggtc ttggtaagac ctggtcaata
     1801 tttcttcgat tttctgacga tctttaaggg cttcactatt cgttaaatca gccggcattt
     1861 tcccagattc agatttttgc cttagctgga aaattgtatt tatagcatca ttgtattttt
     1921 ggacatcagg ggtggatttc aaagtcttgt caaatgcagc gcagaggcta tcaatagtag
     1981 agctatcgat tggatcattc cctttctcac acagtggcat cgtgattcta attttttagt
     2041 aatcttttca tgtgagtagt ttagataaaa cgtcttcact attaagtaga agattcgtaa
     2101 ttggatttaa gccttcgtgc cttatcgcgg ctgataatca tcatgatcga cgatgaacta
     2161 ctgcgccaga gtataatccc ctataaaaca ggcctgctat cactccaaag agaatgcttt
     2221 tatagaagaa tataacgtaa attactacaa taattgtgtt gagatgtcta aaggtgaaga
     2281 attattcact ggtgttgtcc caattttggt tgaattagat ggtgatgtta atggtcacaa
     2341 attttctgtc tccggtgaag gtgaaggtga tgctacttac ggtaaattga ccttaaaatt
     2401 tatttgtact actggtaaat tgccagttcc atggccaacc ttagtcacta ctttaactta
     2461 tggtgttcaa tgtttttcta gatacccaga tcatatgaaa caacatgact ttttcaagtc
     2521 tgccatgcca gaaggttatg ttcaagaaag aactattttt ttcaaagatg acggtaacta
     2581 caagaccaga gctgaagtca agtttgaagg tgatacctta gttaatagaa tcgaattaaa
     2641 aggtattgat tttaaagaag atggtaacat tttaggtcac aaattggaat acaactataa
     2701 ctctcacaat gtttacatca tggctgacaa acaaaagaat ggtatcaaag ttaacttcaa
     2761 aattagacac aacattgaag atggttctgt tcaattagct gaccattatc aacaaaatac
     2821 tccaattggt gatggtccag tcttgttacc agacaaccat tacttatcca ctcaatctgc
     2881 cttatccaaa gatccaaacg aaaagagaga ccacatggtc ttgttagaat ttgttactgc
     2941 tgctggtatt acccatggta tggatgaatt gtacaaagct tcgtacgctg caggtcgacg
     3001 gatccccggg ttaattaaca tgtcggaaga cgaatttttt ggtggtgata atgaagccgt
     3061 ttggaacggt tccagattca gtgattcacc tgagttccaa acgttgaaag aagaggttgc
     3121 tgcagagtta tttgaaataa atgggcaaat aagcacgctg cagcagttta ccgcgacact
     3181 taagtcattt atagatcggg gagatgttag tgcgaaagtt gtggaaagaa ttaataagag
     3241 atctgtggca aagatagaag aaataggcgg gctcattaaa aaggtcaata catcagtgaa
     3301 aaagatggat gcgattgagg aagctagcct ggataagact caaataatag cgagagagaa
     3361 acttgtgagg gatgtcagtt actcttttca agagtttcaa ggtattcagc ggcagtttac
     3421 ccaagtaatg aaacaagtta atgaaagagc aaaagaatct cttgaagcaa gtgagatggc
     3481 aaatgatgct gctttattag atgaagaaca aaggcagaat agctcaaaaa gtactcgaat
     3541 accaggcagc caaatagtca ttgagagaga cccgataaat aacgaagagt ttgcttatca
     3601 gcaaaatctt atcgagcaaa gagaccagga aatcagcaat attgaaagag gtataacgga
     3661 actgaacgaa gtttttaaag atttgggaag tgttgttcaa cagcagggtg tgctagttga
     3721 caatattgaa gcaaatattt atacaacgtc agataacact caattggctt cagacgagct
     3781 aaggaaggcc atgcggtacc aaaaacgtac gagcagatgg agggtgtatt tgttgattgt
     3841 gcttctcgta atgcttcttt ttatttttct cattatgaaa ttgtaagcgg ccgcaaaaaa
     3901 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
     3961 atcagttatt acccgggaat ctcggtcgta atgatttcta taatgacgaa aaaaaaaaaa
     4021 ttggaaagaa aaagcttcat ggcctttata aaaaggaact atccaatacc tcgccagaac
     4081 caagtaacag tattttacgg ggcacaaatc aagaacaata agacaggact gtaaagatgg
     4141 acgcattgaa ctccaaagaa caacaagagt tccaaaaagt agtggaacaa aagcaaatga
     4201 aggatttcat gcgtttgtac tctaatctgg tagaaagatg tttcacagac tgtgtcaatg
     4261 acttcacaac atcaaagcta accaataagg aacaaacatg catcatgaag tgctcagaaa
     4321 agttcttgaa gcatagcgaa cgtgtagggc agcgtttcca agaacaaaac gctgccttgg
     4381 gacaaggctt gggccgataa ggtgtactgg cgtatatata tctaattatg tatctctggt
     4441 gtagcccatt tttagcatgt aaatataaag agaaaccata tctaatctaa ccaaatccaa
     4501 acaaaattca atagttacgt agacgtccta cgattccgcg gtggagctcc agcttttgtt
     4561 ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4621 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4681 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4741 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4801 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4861 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4921 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4981 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5041 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5101 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5161 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5221 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5281 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5341 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5401 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5461 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5521 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5581 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5641 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5701 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5761 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5821 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5881 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5941 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6001 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6061 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6121 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6181 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6241 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6301 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6361 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6421 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6481 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6541 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6601 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6661 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6721 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     6781 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     6841 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     6901 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     6961 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     7021 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     7081 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     7141 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     7201 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     7261 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     7321 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga
     7381 ctcactatag ggcgaattgg gtacccgttg ctaaattcga gtgaaacaca ggaagaccag
     7441 aaaatcctca tttcatccat attaacaata atttcaaatg tttatttgca ttatttgaaa
     7501 ctagggaaga caagcaacga aacgtttttg aaaattttga gtattttcaa taaatttgta
     7561 gaggactcag atattgaaaa aaagctacag caattaatac ttgataagaa gagtattgag
     7621 aagggcaacg gttcatcatc tcatggatct gcacatgaac aaacaccaga gtcaaacgac
     7681 gttgaaattg aggctactgc gccaattgag gccttttttg cgaggcatat ttatggtgaa
     7741 ggataagttt tgaccatcaa agaaggttaa tgtggctgtg gtttcagggt ccataaagct
     7801 t
//
