LOCUS       sequence-292109-        7553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of CHS5-mCherry as an additional copy in genome, use 
            auxotrophic marker TRP1(Kluyveromyces lactis). Marker for yeast late
            golgi/early endosome..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7553)
  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 7553)
  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: ZJOM73
FEATURES             Location/Qualifiers
     source          1..7553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             3106..3816
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3251..3269)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3519..3538
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      3846..4033
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3901..3925)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    4085..4118
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5121..5154
                     /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(5207..5226)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5208..5226)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5313..5330)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5484..6072)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5564..5583)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6243..7034,7035..7103))
                     /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     6866..6885
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7104..7208)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7276..7294
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7332..7354)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7454..7473
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaggtga cactatagaa cgcggccgcc agctgaagct tctaactata ataactggta
       61 gctttgtcac tcgtaccagg aaaagtgaag attaaactga attttaaaat gaacgatcga
      121 ttagttacgg aagagcaaga gttgtttaca aaattgcgcg agattgtagg ttcaagtatt
      181 cgcttttggg aggaacaact gttttatcaa gttcaagatg taagcaccat agaaaaccac
      241 gtcattctca gtttaaaatg tacaatttta acggatgctc agataagtac gttcataagc
      301 aaacccagag agcttcatac gcatgccaaa ggatatcctg aaatctatta cctttccgag
      361 ttatcaacaa ctgtcaattt tttttctaaa gagggaaact atgtcgaaat aagccaggtt
      421 attcctcatt ttaatgaata tttttcctct ttaatagtgt ctcaattgga atttgaatac
      481 ccgatggtct tctccatgat ttcaaggctc cgattgaagt ggcaacaaag ttcgctcgct
      541 ccgatatcct acgccctaac gagcaattca gtacttcttc caataatgct taacatgatt
      601 gcccaagaca aatcttcaac aaccgcgtat caaattctgt gtcgaagaag aggtcctcca
      661 attcagaatt ttcaaatttt ttccttaccg gctgtaacgt acaataagta gcatgcataa
      721 aatataattt aatcaaatac ttttgggcaa ttaaaatttt agttaacaat agttatgcaa
      781 tgcgctttat gttcatatga taccgtttat aagctattgc catatcctta tcttattgct
      841 tccagtagcc tcgagtcgac cactaaaaag atgtcactta agacggaaat tatgtagctg
      901 cacttctttt ttaacaagtt cggtcggccc ttcaagttct cctttctaaa gcctcattat
      961 ttattgcgta gatgctaaat gttatcgcgg tttagcttgc atgttacgtt tccgttttag
     1021 aacctggtcg agtagcgaat aatgtcttca gttgatgtac tgttaacagt aggtaagttg
     1081 gatgcctcat tggcgttact gactactcag gatcatcatg ttattgagtt tcctacagta
     1141 ttattaccag aaaatgttaa agctggatct atcataaaaa tgcaagtttc acaaaattta
     1201 gaggaggaaa aaaaacaaag gaatcatttt aagagtatac aagccaaaat tttggagaag
     1261 tatggtaccc ataaaccgga gagcccagtt ttgaaaattg ttaacgttac gcaaacgagc
     1321 tgtgttctag catgggatcc attgaaactt ggctcagcaa aattgaaatc actgatcctt
     1381 tataggaagg gaatacgttc aatggtaatt ccaaatccat tcaaagtgac taccacgaaa
     1441 atatccggtc tttccgttga tacgccatac gaatttcaat tgaaactgat aaccacgtca
     1501 ggaacattat ggtctgaaaa ggttatattg cgtacacata agatgactga catgtctggt
     1561 atcactgtat gtttgggtcc attggatcca ttgaaagaaa tttcagactt acagatatcc
     1621 caatgtttgt ctcacatcgg ggcgagacct ttacaacgtc atgttgcgat agatactacg
     1681 cattttgtct gtaacgatct agacaatgaa gaaagcaatg aagagcttat aagggcaaaa
     1741 cataacaaca taccaattgt cagaccggaa tgggtgagag cttgtgaggt tgagaaaaga
     1801 atcgttggtg ttagaggatt ttacttagat gcagatcaaa gtatactgaa aaactacaca
     1861 ttcccaccag ttaatgagga agaactttcg tactcaaagg agaatgagcc ggtagccgaa
     1921 gtagcggatg aaaataagat gcccgaggac acaacagatg tcgaacaggt tgcatcacct
     1981 aatgacaatg agagtaatcc ttcagaagct aaggaacaag gagaaaagag tggacatgaa
     2041 actgccccag taagtcctgt agaagatcca ttgcatgctt cgacggcttt ggagaatgaa
     2101 accaccatcg aaaccgtcaa cccctccgta agaagtttga aaagcgaacc tgttggtact
     2161 cccaatatag aggaaaacaa agcggactct tccgcagaag ccgtggtaga agaaccgaat
     2221 gaagctgtgg ctgaaagttc tccaaatgaa gaagcaacgg gacagaaaag tgaggatacc
     2281 gatacacatt ctaacgaaca agctgataat ggatttgtac agactgaaga agtagctgaa
     2341 aacaacataa ccacagaaag tgcaggggaa aataacgaac ctgcagatga tgcagcaatg
     2401 gaatttggac gtccagaagc tgaaattgaa actccagaag taaatgagtc tatagaagat
     2461 gccaatgaac ctgcggagga ttccaatgaa cctgtggagg attccaacaa acctgtgaag
     2521 gattccaaca aacctgtgga ggattccaac aaacctgtgg aggattccaa caaacctgtg
     2581 gaggattcca acaaacctgt ggaggatgcc aatgaacctg tggaagatac cagtgaacct
     2641 gtggaggatg ccggtgaacc cgtacaagaa accaacgagt ttactaccga cattgcctct
     2701 ccaagacatc aagaagaaga tatagaactt gaagccgaac ctaaagatgc taccgaaagt
     2761 gttgcagtcg agccatccaa tgaagatgta aaaccagaag aaaaaggttc agaggcagaa
     2821 gacgatatca acaacgtttc caaggaggct gcctctggtg agagtactac ccaccaaaaa
     2881 actgaggcct ctgcttctct tgaaagcagt gccgtcacgg aagaacaaga gacaacggaa
     2941 gccgaagtaa atacagatga cgttttgtcc actaaagaag ctaaaaaaaa tactggcaac
     3001 agcaacagta ataagaagaa gaataagaag aataagaaga aagggaaaaa gaaagcttcg
     3061 tacgctgcag gtcgtgggat tcctgggtta attaacatcg ctagcatggt gagcaagggc
     3121 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     3181 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     3241 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     3301 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     3361 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     3421 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     3481 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     3541 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     3601 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     3661 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     3721 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     3781 cactccaccg gcggcatgga cgagctgtac aagtagccta ggtgaggcgc gccacttcta
     3841 aataagcgaa tttcttatga tttatgattt ttattattaa ataagttata aaaaaaataa
     3901 gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta ttcttgagta
     3961 actctttcct gtaggtcagg ttgctttctc aggtatagta tgaggtcgct cttattgacc
     4021 acacctctac cggcagatcc gctagggata acagggtaat atctgcaggt cgacaaccct
     4081 taatataact tcgtataatg tatgctatac gaagttatta ggtctagagc ggccgcggaa
     4141 cgatcattca ctatatatat atcaatttat atatacgtat gtgtaattga agaaagatac
     4201 gttttttcct ctattgagag gcctgctgga tgaatagctt taccttttct aaatccttga
     4261 taccatcagt ctctactcct ccacttacat cgacaccgat cgcatttggt aacatattaa
     4321 tggcaacaga aacgttatca ggattcaatc caccagcgat aatgaatttt atctcgggat
     4381 gacttgcaga ccaactggaa attgcactcc aattcaattt ctcaccagtg ccaccttcac
     4441 cagaatcgaa caacgtcagc acattgtcta cgtgttcata caggtccagt agtaattcac
     4501 aatcctgtgg gaactggaac ctcttaatga ttggaattga agatgggatc aaagatctgt
     4561 attctttaat atcttcatct ccatgtaatt gtatcacatc tagattatat tcgtggtaca
     4621 gttgaaggac atcatcaacg gactgatttc taaacacccc gactagttta gtacctttca
     4681 cgttctcttg ttggtgaact gcagttgaaa tacctttcgc aacagatgag tcaatggttc
     4741 ttttcctacc gggaacacaa atgataccta agtaatcagc accatcatcc acagcagtct
     4801 ttgcagcttc aacggtttgc aaaccacaca ctttaacgag catcacagta gatttcgcaa
     4861 gagggtagct cgctcagagt acccaagtaa atgattagta aactgatgtt tgatagttca
     4921 atttttcaat gaaataacct tatattaaaa ttgatattac tattatacaa aaataaagaa
     4981 taaaggattt gagtttatac ataaaatacc attattattt gttcagtgag agatactggg
     5041 gtatatggga tgtgtgtagt gataccatgc aatcatgtat caaacatggg cccggtaccg
     5101 agctctcgag aacccttaat ataacttcgt ataatgtatg ctatacgaag ttattaggtg
     5161 atatcagatc cactagtggc ctatgcggcc gcggatctgc cggtctccct atagtgagtc
     5221 gtattaattt cgataagcca ggttaacctg cattaatgaa tcggccaacg cgcggggaga
     5281 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5341 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5401 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5461 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5521 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5581 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5641 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5701 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5761 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5821 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5881 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5941 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6001 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6061 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6121 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6181 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6241 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6301 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6361 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6421 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6481 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6541 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6601 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6661 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6721 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6781 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6841 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6901 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6961 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7021 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7081 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7141 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7201 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     7261 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     7321 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     7381 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     7441 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat ggacatattg
     7501 tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat acg
//
