LOCUS       sequence-265901-        7300 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of VPS4-GFP as an additional copy in genome, use 
            auxotrophic marker URA3(Candida albicans). Marker for yeast late 
            endosome..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7300)
  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 7300)
  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: ZJOM10
FEATURES             Location/Qualifiers
     source          1..7300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2385..3095
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(2412..2440)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3035..3055
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3125..3312
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3180..3204)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    3364..3397
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4849..4882
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(4935..4954)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4936..4954)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5041..5058)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5212..5800)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5292..5311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5971..6762,6763..6831))
                     /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     6594..6613
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6832..6936)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7004..7022
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7060..7082)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7182..7201
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7282..7300
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7282..7299
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttcatcactt ttgttccttt tgtcgttttt cttcttttag
       61 attccatctt gaaaagagat gaaatggaca aagtaacata atttagaccc aacaaagtaa
      121 aaagaaggtt gggttggcat tgaagatcac aatgggaatt caggaaaaga cattggggat
      181 acgaaaagag cgcaaattgg tggttgtgcc tcgcgaaaga aatcatgtta gacacgcttc
      241 acaaaggaca cgttctaaaa actacaagaa tatctccaaa aagagggctc aacaacacgc
      301 gtttggattc aatattgcta agactcttgc caagatccaa gcgtttgtat gggggtctcc
      361 cgcagatgag gaagaagaaa gcgttgtccc attgtctaaa aattctcagg attgtgtgcc
      421 tctacaatgg caagcaaagt ttgcgcaatt gcggcagcag ctgcactcta cacaaaagga
      481 actacaattt gtcaaggaaa agtgtcatct gttgcagtcg gtgttggacg acgcaaacat
      541 cgatcaaaga tacctggagt ctcgtcgaga catgaaaaat attgagcgag acaacctcaa
      601 accgacagaa aatttaccac cttcgccagt aagagcagta aacccgttag tgacttcaag
      661 tccgattcac atgtcgccac tccagtcgcg ccaacgacca gtttcttccc tacagccccc
      721 aaagggcccc aacttctacg ccaagtatcc taagcttcct caaacgaaca ttctcagaga
      781 atctccgact gaggattcgg ttccgcacgc tgaatagaaa cacaacacgt catggttgtt
      841 ctttctgtac atctatttat tcttgcttga tgatatttat tgtattgctt tacgggtacc
      901 cccgctgggt gaaaaaaaaa aaaaaataag aacttcatga tatcgaacga tttaatcgtg
      961 ggttgaggac atggaagaca aaaataaagc agcatagagt gcctatagta gatggggtac
     1021 aaatgagcac gggagatttt ttaactaagg gaatagaact ggttcaaaag gccattgacc
     1081 tggacacagc cacgcagtat gaggaggcgt atacagcata ctataacgga ttagattatt
     1141 tgatgttggc tttgaaatac gagaaaaacc ctaagtcgaa ggatttgata agagcgaagt
     1201 ttacagagta tctaaaccgt gcagagcagt tgaaaaagca cttggaaagt gaagaggcaa
     1261 acgcggcaaa gaaatctcct agcgctggta gtggttccaa cggtggtaat aagaaaatct
     1321 ctcaggaaga gggtgaggat aatggtggag aagacaacaa aaagctaaga ggtgccttat
     1381 cgagtgctat tttgtctgag aagcccaatg ttaagtggga agacgtggct ggactggaag
     1441 gagccaaaga ggcccttaaa gaagccgtta ttttacctgt caagttccca catttgttca
     1501 aaggtaaccg taagcccact tcgggaatct tattgtacgg accaccaggt acaggtaaat
     1561 catatttggc aaaagcagtg gccaccgagg ccaattctac ctttttcagt gttagttcca
     1621 gtgatttggt ttctaagtgg atgggtgaat ctgaaaaact tgtcaaacag ttatttgcga
     1681 tggcaagaga aaataaacct tctattattt ttatagatga agtggatgcg ctaacaggta
     1741 ccagaggaga aggagagagc gaagcaagta gaagaattaa aacagaatta ttggtacaaa
     1801 tgaatggtgt tgggaatgac tcccaaggtg ttttggtcct tggcgcaaca aatatcccat
     1861 ggcaactgga cagtgccatc aggagaagat ttgaaagaag aatatatata ccattaccag
     1921 atttggcggc aagaaccacc atgttcgaga ttaacgtcgg cgacactcca tgcgtattga
     1981 ccaaggaaga ttatagaact ttaggtgcaa tgactgaagg ttattccggg agtgatattg
     2041 ctgtagtggt gaaagatgcg ctaatgcaac cgataagaaa aattcaaagc gcgacacatt
     2101 tcaaagatgt ctctacggag gacgacgaaa caagaaaact aacaccatgc tctccaggtg
     2161 atgatggcgc catcgagatg agttggacgg atattgaagc tgatgaatta aaagaaccag
     2221 atctcaccat aaaggatttc ttaaaggcta tcaaatcgac gagacctacc gtgaatgagg
     2281 atgacttgct gaagcaagaa cagttcacta gagattttgg tcaagaaggt aacgcttcgt
     2341 acgctgcagg tcgtgggatt cctgggttaa ttaacatcgc tagcagtaaa ggagaagaac
     2401 ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat
     2461 tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc cttaaattta
     2521 tttgcactac tggaaaacta cctgttccat ggccaacact tgtcactact ttcacttatg
     2581 gtgttcaatg cttttcaaga tacccagatc atatgaaacg gcatgacttt ttcaagagtg
     2641 ccatgcccga aggttatgta caggaaagaa ctatattttt caaagatgac gggaactaca
     2701 agacacgtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc gagttaaaag
     2761 gtattgattt taaagaagat ggaaacattc ttggacacaa attggaatac aactataact
     2821 cacacaatgt atacatcatg gcagacaaac aaaagaatgg aatcaaagtt aacttcaaaa
     2881 ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa caaaatactc
     2941 caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc
     3001 tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg
     3061 ctgggattac acatggcatg gatgaactat acaaacctag gtgaggcgcg ccacttctaa
     3121 ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa aaaaaataag
     3181 tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat tcttgagtaa
     3241 ctctttcctg taggtcaggt tgctttctca ggtatagtat gaggtcgctc ttattgacca
     3301 cacctctacc ggcagatccg ctagggataa cagggtaata tctgcaggtc gacaaccctt
     3361 aatataactt cgtataatgt atgctatacg aagttattag gtctagagat cccaatacaa
     3421 cagatcacgt gatctttttg taagatgaag ttgaagtagt gttgcaccgt gccaatgcag
     3481 gtggctatta gattaaatat gtgatttgtt ctattaagtt tcctgtataa ttaatgggga
     3541 gcgctgattc tcttttggta cgcttcccat ccagcatttc tgtatctttc accttcaacc
     3601 ttaggatctc tacccttggc gaaaagtcct ctgccaacaa tgatgatatc tgatccacca
     3661 cttacaactt cgtcgacggt tctgtactgc tgacccaatg catcgccttt gtcgtctaaa
     3721 cctacacctg gggtcatgat tagccaatca aacccttctt ctcttcctcc catatcgttc
     3781 tgagcaatga acccaataac gaaatcttta tcactctttg caatatcaac ggtaccctta
     3841 gtatattcac cgtgtgctag agaacccttg gaagacaatt cagcaagcat caataatccc
     3901 cttggttctt tggtgacctc ttgcgcacct tgtttcaagc cagcaacaat accagcacca
     3961 gtaaccccgt gggcgttggt gatatcagac cattctgcga tacggtaaac gcccgatgta
     4021 tattgtaatt tgactgtgtt accgatatcg gcgaattttc tgtcctcaaa tatcaagaac
     4081 ttgtatttct ctgccaatgc tttcaatgga acgacagtac cctcataact gaaatcatcc
     4141 aagatatcaa cgtgtgtttt caaaaggcaa atgtatggac ccaacgtttc aacaagtttc
     4201 aatagctcat cagtcgaacg aacgtcaaga gaagcacaca aattggtctt cttttcatcc
     4261 attaaacgta aaagtttcga tgcaaccgga cttgcatgag tctcagctct actggtatat
     4321 gattttgtgg acatggtgca actaattgac gggagtgtat tgacgctggc gtactggctt
     4381 tcacaaaatg gcccaatcac aaccacatct tagatagttg aaatgacttt agataacatc
     4441 aattgagatg agcttaatca tgtcaaagct aaaagtgtca ccatgaacga caattcttaa
     4501 gcaaatcacg tgatatagat ccacgaataa ccaccatttg atgctcgagg caagtaatgt
     4561 gtgtaaaaaa atgcgttacc accatccaat gcagaccgat cttctaccca gaatcacata
     4621 tatttatgta ccgagtacct tttttctatc ttccaattgc ttctcccata tgattgtctc
     4681 cgtaagctcg aaatttctaa gttggatttt aatcttcacg caggatgaca gttcgatgag
     4741 cttctgagga gtgtttagaa cataatcatt tatccatggt ctatctcttc ttgtcgcttt
     4801 ttctcctcga tagaacctaa ataaaacgag ctctcgagaa cccttaatat aacttcgtat
     4861 aatgtatgct atacgaagtt attaggtgat atcagatcca ctagtggcct atgcggccgc
     4921 ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca
     4981 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5041 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5101 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5161 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5221 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5281 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5341 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5401 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5461 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5521 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5581 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5641 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5701 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5761 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5821 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5881 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5941 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6001 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6061 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6121 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6181 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6241 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6301 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6361 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6421 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6481 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6541 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6601 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6661 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6721 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6781 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6841 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6901 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6961 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     7021 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     7081 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     7141 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     7201 actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac
     7261 cttatgtatc atacacatac gatttaggtg acactataga
//
