LOCUS       sequence-269552-        6685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of ERG6-mCherry as an additional copy in genome, use 
            auxotrophic marker TRP1(Kluyveromyces lactis). Marker for yeast 
            lipid droplet..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6685)
  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 6685)
  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: ZJOM70
FEATURES             Location/Qualifiers
     source          1..6685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2225..2935
                     /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(2370..2388)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2638..2657
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      2965..3152
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3020..3044)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    3204..3237
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4240..4273
                     /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(4326..4345)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4327..4345)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4432..4449)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4603..5191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4683..4702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5362..6153,6154..6222))
                     /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     5985..6004
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6223..6327)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6395..6413
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6451..6473)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6573..6592
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(6673..6685,1..6)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(6673..6685,1..5)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 tatagaacgc ggccgccagc tgaatgctcg ctatcctcgc catcacgtgt accagtacgt
       61 ctatgttcat tttctatttc atcgtgacga gattttgaac cctcaaactc cgctaatgaa
      121 cccggagaaa cgacatccag cgacctcttg gcggtagaca cactcatggt ttaagaaaca
      181 acttttcctt ctttaaacgt tttgggtggc aaaaaacttc ccttaaacaa actaaaaatg
      241 gctcgtgttc atgcacaagt caaccagggg ctcaggaaaa aaagctaaac accaatccta
      301 tatataacgt aaaggtactg tgcttaatcg gtaaacggaa gactattaag caaatgtatc
      361 tcttaagacc ttacgcatcc atcagttgcc accctcctac tcttcatcgg agggtttccg
      421 gttcccatga caaagacggc gctttggcgg ggcccaaatt gttacccgca cccagaaaaa
      481 gaaaagagta acctaggtca aatcaaccaa acagctaggg ctggttacta tttttgaaaa
      541 gatgcaacag ggtaagatca gcaaaattta gggccgaaca aagccccgat cttcgtatat
      601 ggtacctcgt tcccgtactt attttcaatt tttggcgtcg tttactgccg tttccgataa
      661 atacaaaaaa gttttcgaga agcgagagta agtgaggagc agacaatgta ttattcactg
      721 taaactgctc cacttcgtct caatggtaca cttcattttt atcgccttac gaagcatgcg
      781 ttttatgcga agattggtga gaaacctcca atacttgctg ttgccgataa cttcttcatt
      841 gctttttata tagttcgggt gttttctcct atcctctgct gctctctttt tctttccatt
      901 cgtctttgta atttataagc tgttaatttt cattcaagtt tcccttatct gttttacttt
      961 cgatttaagt tttacataat ttaaaaaaac aagaataaaa taataatata gtaggcagca
     1021 taagatgagt gaaacagaat tgagaaaaag acaggcccaa ttcactaggg agttacatgg
     1081 tgatgatatt ggtaaaaaga caggtttgag tgcattgatg tcgaagaaca actctgccca
     1141 aaaggaagcc gttcagaagt acttgagaaa ttgggatggt agaaccgata aagatgccga
     1201 agaacgtcgt cttgaggatt ataatgaagc cacacattcc tactataacg tcgttacaga
     1261 tttctatgaa tatggttggg gttcctcttt ccatttcagc agattttata aaggtgagag
     1321 tttcgctgcc tcgatagcaa gacatgaaca ttatttagct tacaaggctg gtattcaaag
     1381 aggcgattta gttctcgacg ttggttgtgg tgttgggggc ccagcaagag agattgcaag
     1441 atttaccggt tgtaacgtca tcggtctaaa caataacgat taccaaattg ccaaggcaaa
     1501 atattacgct aaaaaataca atttgagtga ccaaatggac tttgtaaagg gtgatttcat
     1561 gaaaatggat ttcgaagaaa acactttcga caaagtttat gcaattgagg ccacatgtca
     1621 cgctccaaaa ttagaaggtg tatacagcga aatctacaag gttttgaaac cgggtggtac
     1681 ctttgctgtt tacgaatggg taatgactga taaatatgac gaaaacaatc ctgaacatag
     1741 aaagatcgct tatgaaattg aactaggtga tggtatccca aagatgttcc atgtcgacgt
     1801 ggctaggaaa gcattgaaga actgtggttt cgaagtcctc gttagcgaag acctggcgga
     1861 caatgatgat gaaatccctt ggtattaccc attaactggt gagtggaagt acgttcaaaa
     1921 cttagctaat ttggccacat ttttcagaac ttcttacttg ggtagacaat ttactacagc
     1981 aatggttact gtaatggaaa aattaggtct agccccagaa ggttccaagg aagttactgc
     2041 tgctctagaa aatgctgcgg ttggtttagt tgccggtggt aagtccaagt tattcactcc
     2101 aatgatgctt ttcgtcgcta ggaagccaga aaacgccgaa accccctccc aaacttccca
     2161 agaagcaact caagcttcgt acgctgcagg tcgtgggatt cctgggttaa ttaacatcgc
     2221 tagcatggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
     2281 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
     2341 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
     2401 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
     2461 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
     2521 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
     2581 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     2641 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
     2701 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     2761 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     2821 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     2881 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtagcctag
     2941 gtgaggcgcg ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa
     3001 taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga
     3061 aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat
     3121 gaggtcgctc ttattgacca cacctctacc ggcagatccg ctagggataa cagggtaata
     3181 tctgcaggtc gacaaccctt aatataactt cgtataatgt atgctatacg aagttattag
     3241 gtctagagcg gccgcggaac gatcattcac tatatatata tcaatttata tatacgtatg
     3301 tgtaattgaa gaaagatacg ttttttcctc tattgagagg cctgctggat gaatagcttt
     3361 accttttcta aatccttgat accatcagtc tctactcctc cacttacatc gacaccgatc
     3421 gcatttggta acatattaat ggcaacagaa acgttatcag gattcaatcc accagcgata
     3481 atgaatttta tctcgggatg acttgcagac caactggaaa ttgcactcca attcaatttc
     3541 tcaccagtgc caccttcacc agaatcgaac aacgtcagca cattgtctac gtgttcatac
     3601 aggtccagta gtaattcaca atcctgtggg aactggaacc tcttaatgat tggaattgaa
     3661 gatgggatca aagatctgta ttctttaata tcttcatctc catgtaattg tatcacatct
     3721 agattatatt cgtggtacag ttgaaggaca tcatcaacgg actgatttct aaacaccccg
     3781 actagtttag tacctttcac gttctcttgt tggtgaactg cagttgaaat acctttcgca
     3841 acagatgagt caatggttct tttcctaccg ggaacacaaa tgatacctaa gtaatcagca
     3901 ccatcatcca cagcagtctt tgcagcttca acggtttgca aaccacacac tttaacgagc
     3961 atcacagtag atttcgcaag agggtagctc gctcagagta cccaagtaaa tgattagtaa
     4021 actgatgttt gatagttcaa tttttcaatg aaataacctt atattaaaat tgatattact
     4081 attatacaaa aataaagaat aaaggatttg agtttataca taaaatacca ttattatttg
     4141 ttcagtgaga gatactgggg tatatgggat gtgtgtagtg ataccatgca atcatgtatc
     4201 aaacatgggc ccggtaccga gctctcgaga acccttaata taacttcgta taatgtatgc
     4261 tatacgaagt tattaggtga tatcagatcc actagtggcc tatgcggccg cggatctgcc
     4321 ggtctcccta tagtgagtcg tattaatttc gataagccag gttaacctgc attaatgaat
     4381 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4441 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4501 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4561 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4621 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4681 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4741 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4801 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4861 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4921 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4981 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5041 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5101 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5161 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5221 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5281 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5341 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5401 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5461 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5521 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5581 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5641 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5701 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5761 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5821 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5881 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5941 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6001 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6061 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6121 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6181 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6241 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6301 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     6361 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     6421 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     6481 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     6541 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     6601 gcaccatatg gacatattgt cgttagaacg cggctacaat taatacataa ccttatgtat
     6661 catacacata cgatttaggt gacac
//
