LOCUS       sequence-268966-        7466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of TGL3-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 7466)
  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 7466)
  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: ZJOM68
FEATURES             Location/Qualifiers
     source          1..7466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     83..101
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(139..161)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     261..280
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        361..379
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     361..378
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             3379..4089
                     /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(3524..3542)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3792..3811
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      4119..4306
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4174..4198)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    4358..4391
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5394..5427
                     /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(5480..5499)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5481..5499)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5586..5603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5757..6345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5837..5856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6516..7307,7308..7376))
                     /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     7139..7158
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7377..7466,1..15))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
       61 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
      121 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
      181 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
      241 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgga
      301 catattgtcg ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg
      361 atttaggtga cactatagaa cgcggccgcc agctgaagct tgactggctg atagcctcga
      421 cgccagcttt gattagttca tccgggttcc cgtcaatttt tataaacgtg tctaaagttc
      481 tctctagata agtctttgcc ccttggcttc ttgcgccaat agctgtacca tatagttctg
      541 tgacatttcc ggaaggttgg aattctaaaa gatgagcacc gctcttgtca tacccgatga
      601 tcaacaaacc aacaccgtag ggtctgcccc cataggattg tgtatttttc tgtgctttat
      661 cgcaaagcaa atgacctgct ctctcgacgg caagctttct gttaaaaact aagctggagt
      721 aattacactg ttgtctcaaa taattactta aaactctagc atctggagcc aacccggcta
      781 acgagagccc catgtgctcg tcacatttga tgattttttt ctggtatgat gacaattcgt
      841 ctgcatttct tttcaacgct actaaaacag catgagtgtt cgaacgtaac cccacagtga
      901 cgctcccttg tttaatagct tccaaggcgt attccacttg gaacagacgg cctgtaggag
      961 aaaaagtgac tgtgtccccg tcgtaattgt tcctgaacat gttttgtaaa tatatgaagg
     1021 cttagttcaa tgtattctta cccacttctc ttgaacttca attgatcacc acgtctacct
     1081 aaatgtagtg gcgtcttggt tcttttccat actttgacga tacttttgaa tttttaattt
     1141 tgccactcta taatagggta tgttctttat aggtcatttt tactttttta gtagggcatt
     1201 ttaataaata ctaggttttt agtaatttcg gagatactta tcctaggtct gaaaattcaa
     1261 ccctaaccct ttttttctta tgtttttttt tttctgggta gcagctaata gaaattagta
     1321 ctgattacat agagaagtac tatgacacaa tagtaaggga atcatctatt catatatcac
     1381 atctttgagt tgccgttaag catgaaggaa acggcgcagg aatacaaggt gtctgctgta
     1441 ataccgaccc ttttgaaaaa ctggatactg cgtgtagtgt atgccacatt ggaccatata
     1501 cctccttttg tgtgggaaat tctacatgtc atcaccgata tttatttctt ctgggttcaa
     1561 aagctgatta actatgtgcg gccacactca agagtcattt attataacgc tattaagaag
     1621 ttagatgagt gtgatacgta tcaaatgtgg tgtcagcaag cgtccgtagt ggatgaaata
     1681 acaggcgcaa atttatggcg gcggaatttc ttttcgagga ggtacgactt caattctgtc
     1741 attgaacaat atagcatatt ggaaaatatg ttaagggagg agaagtatga tgtagttaag
     1801 gagaagtttt cgactacagg cccttgtatg ttgaggaact ttgcgggtat tggagataaa
     1861 aagttgttca caaaatcttt gatgggtaca aagctactta ttgaacaata tttgacccga
     1921 attctagaag gcttggatat attaaataat caaactttaa ctccaacatc ctttttccaa
     1981 agatgtaagt tgtcactggg cacaacagca ttaatcttgc aaggtggttc attgtttggg
     2041 ctgtttcacc ttggggtgat tagaggttta ttgttacagg acctgatgcc taatattata
     2101 agtggtagct ctatgggggc atgcgttgca agtttattcg gttgcttgtc caatgagcaa
     2161 ctaaagcaat tattgaccga tgacaacctc ttgaatatca tcaagaatga tgtagattta
     2221 ttaaaaagct gtggatacgg taatttagag cagcacctaa acctggggac tttgattcag
     2281 aacctaatac accacggtta ttctcaagac gtttatcttt ttatccggtt tgttatgaaa
     2341 tatattgtta aggaaaaaac ttttgaagaa gtttatcaaa ttactggaaa agttttcaat
     2401 attgttattc atccaactga taaatcatgc cctaatttac taaattatgt gacaacccca
     2461 aacgtattaa tcaagtcagc cattgaatgc tcacttggtt ctggtgtaat ttcagaggat
     2521 acatcactat tatgtaaaaa tttagaaaat gaaattgagc cttttcttaa catcaataaa
     2581 aataaacaag tcaaatttct taccccagaa aatgccaaca atccgagcat aacggagagc
     2641 ccctatacta ggttaacgga actattcaat gttaataact tcattgtttc tttggcgaga
     2701 ccatacttgg caccattggt ggtaaatgat ttgaagcatg aaataaaaac gtcaaaatat
     2761 tactattata agcattatcc aaatatgcct cctataaacg cgaatacagt aagaaaaaca
     2821 cagagatcct cttcgcaatc acccataaag gcaggaactg tcgaggatct tgagccagag
     2881 cctttaatga gccctgtgcc cccaagttcg gcagtcaacg attcagcaga gtacataatc
     2941 ccagaattgg gcataccgca gttgaacttt actgaaatgg aaccgctggc gtttaaattt
     3001 aagtatcatc tggaaagaaa attaaaaaat attgccacta tggaatttcg acataggatg
     3061 gaggtattgg ataatttagg cctgttgtgc tcattaataa aaagactaat aattgatgaa
     3121 aagactccca gatcagctac ggagattgct gtggtaccaa ggatgaaaag tctatcatta
     3181 acaaggatta ttgaaggtca actaaacaac attccatact ggataaaatc tggtgaacga
     3241 agtacatggc ccgcactagc tttaattaag acaagatgtg cagtcgaatt taaattagac
     3301 gacataataa gagcaagacg gagtagggct tcgtacgctg caggtcgtgg gattcctggg
     3361 ttaattaaca tcgctagcat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
     3421 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
     3481 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     3541 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
     3601 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
     3661 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
     3721 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     3781 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
     3841 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     3901 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     3961 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     4021 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     4081 tacaagtagc ctaggtgagg cgcgccactt ctaaataagc gaatttctta tgatttatga
     4141 tttttattat taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt
     4201 aggttttaaa acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt
     4261 ctcaggtata gtatgaggtc gctcttattg accacacctc taccggcaga tccgctaggg
     4321 ataacagggt aatatctgca ggtcgacaac ccttaatata acttcgtata atgtatgcta
     4381 tacgaagtta ttaggtctag agcggccgcg gaacgatcat tcactatata tatatcaatt
     4441 tatatatacg tatgtgtaat tgaagaaaga tacgtttttt cctctattga gaggcctgct
     4501 ggatgaatag ctttaccttt tctaaatcct tgataccatc agtctctact cctccactta
     4561 catcgacacc gatcgcattt ggtaacatat taatggcaac agaaacgtta tcaggattca
     4621 atccaccagc gataatgaat tttatctcgg gatgacttgc agaccaactg gaaattgcac
     4681 tccaattcaa tttctcacca gtgccacctt caccagaatc gaacaacgtc agcacattgt
     4741 ctacgtgttc atacaggtcc agtagtaatt cacaatcctg tgggaactgg aacctcttaa
     4801 tgattggaat tgaagatggg atcaaagatc tgtattcttt aatatcttca tctccatgta
     4861 attgtatcac atctagatta tattcgtggt acagttgaag gacatcatca acggactgat
     4921 ttctaaacac cccgactagt ttagtacctt tcacgttctc ttgttggtga actgcagttg
     4981 aaataccttt cgcaacagat gagtcaatgg ttcttttcct accgggaaca caaatgatac
     5041 ctaagtaatc agcaccatca tccacagcag tctttgcagc ttcaacggtt tgcaaaccac
     5101 acactttaac gagcatcaca gtagatttcg caagagggta gctcgctcag agtacccaag
     5161 taaatgatta gtaaactgat gtttgatagt tcaatttttc aatgaaataa ccttatatta
     5221 aaattgatat tactattata caaaaataaa gaataaagga tttgagttta tacataaaat
     5281 accattatta tttgttcagt gagagatact ggggtatatg ggatgtgtgt agtgatacca
     5341 tgcaatcatg tatcaaacat gggcccggta ccgagctctc gagaaccctt aatataactt
     5401 cgtataatgt atgctatacg aagttattag gtgatatcag atccactagt ggcctatgcg
     5461 gccgcggatc tgccggtctc cctatagtga gtcgtattaa tttcgataag ccaggttaac
     5521 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5581 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5641 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5701 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5761 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5821 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5881 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5941 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6001 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6061 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6121 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6181 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6241 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6301 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6361 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6421 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6481 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6541 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6601 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6661 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6721 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6781 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6841 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6901 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6961 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7021 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7081 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7141 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7201 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7261 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7321 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7381 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7441 tttgaatgta tttagaaaaa taaaca
//
