LOCUS       sequence-267529-        8060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of VPH1-mCherry as an additional copy in genome, use 
            auxotrophic marker TRP1(Kluyveromyces lactis). Marker for yeast 
            vacuole..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8060)
  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 8060)
  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: ZJOM65
FEATURES             Location/Qualifiers
     source          1..8060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3594..4304
                     /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(3739..3757)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4007..4026
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      4334..4521
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4389..4413)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    4573..4606
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5609..5642
                     /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(5695..5714)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5696..5714)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5801..5818)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5972..6560)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6052..6071)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6731..7522,7523..7591))
                     /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     7354..7373
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7592..7696)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7764..7782
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7820..7842)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7942..7961
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        8042..8060
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     8042..8059
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc tttgtggtgt tgcgttaggt ctaaataacc tagttcccag
       61 gttgaaaaac atttctccac attcaaagtt gatcttgggc cgtttggtgc catttgccgc
      121 cgtggtcagt gccggtattg tcaacgtctt cctaatgaga ggcaacgaaa ttagaaaggg
      181 tatttctgtg tttgactcta acggtgacga agtcggaaag tccaagaagg ctgccttcat
      241 ggcagttggt gaaactgcat taagtagagt gattaacgcc actccaacta tggtcatccc
      301 acctttgatt ttggttagac tacaacgcgg tgtcctcaaa ggtaagtctc tcggtgtgca
      361 aactttggcc aatttggggt tgatctcagt cacaatgttc tcagctttgc catttgcact
      421 cggtatcttc cctcaaagac aagcaatcca tttaaataaa ctagaacctg aactacatgg
      481 caagaaggat aaggatggga agccaattga aaaggtctat tttaacagag gtatttagaa
      541 gtgaagagac aaatttatat agttatagaa taaatatcag atagataaga aaaaaaaatt
      601 agttaaacat taatatatat atatgtgtag tgactgacat acgtatgact gctagtaatc
      661 cagttgccga gctattgttg cagattgaaa tgtcatcacg tggacattat atcatcatta
      721 acgactaatg cgtcaaaaaa aaaatgggct aaaaaaaaaa aaagcaaaaa aaagaaactc
      781 agataagtcg atatggaaaa ccttttttct tgagtgcaac ccttttagag gttacaaaac
      841 aacaaaagtc caaataagtc catagctttt gcctctcaat atatctatac ttgcttagag
      901 ggctacctgt gtgtatttgc tagcttttac ctttctctat acaacaagtg agggtttgaa
      961 cttttaaatc ccttttaatt tgaatcaaaa aaaaaacatt taaaggttac acaaggaaaa
     1021 taatggcaga gaaggaggaa gcgatttttc gctctgctga aatggcttta gtccaattct
     1081 atattcctca agaaatttca agagactctg cttacacttt aggtcaattg ggtcttgttc
     1141 aattccgtga cttgaactct aaggtgcgtg cgtttcaaag aactttcgtg aacgaaatta
     1201 gaagactgga taatgtagaa agacagtatc gttactttta ttctcttttg aagaaacacg
     1261 atattaagct ctacgaagga gacacggaca aatatttgga cggctcaggt gaattgtacg
     1321 ttccaccaag cggttcagtg atagatgatt atgtccggaa cgcttcatat ttggaagaaa
     1381 gattgattca aatggaggat gcaaccgatc aaattgaagt ccagaaaaat gacttggaac
     1441 agtatcgctt tattttgcag tcaggtgatg aatttttctt gaagggtgat aataccgaca
     1501 gcacttccta tatggatgaa gacatgatcg acgctaatgg ggaaaacatt gctgctgcta
     1561 tcggtgcttc tgtaaactat gtcactggtg tcattgctag agacaaagtt gccaccttag
     1621 aacaaattct ttggagagta ttaagaggta accttttctt caaaactgtt gaaattgaac
     1681 aacctgttta tgatgtcaaa accagggagt ataaacataa aaatgctttt atcgtatttt
     1741 ctcacggtga tctgattatt aaaagaatca gaaagattgc ggaatcattg gatgccaatc
     1801 tttacgatgt tgactcttcc aacgagggta gatcacaaca attggccaag gtcaacaaga
     1861 atttgagtga tttgtacaca gttttgaaaa ccacttctac cactttagaa agtgaattat
     1921 atgccattgc caaagaattg gactcttggt tccaagatgt tacccgtgaa aaggcgattt
     1981 ttgaaatttt gaacaagtct aactatgata ccaatagaaa gattttgatt gctgaaggtt
     2041 ggataccaag agacgaattg gctactttgc aagctcgtct tggtgaaatg atcgcaagat
     2101 tgggtattga tgtcccatcc attatccaag tcctggatac aaaccacact ccacctacct
     2161 tccacagaac taacaagttt actgctggtt tccaaagtat ctgtgactgt tacggtattg
     2221 ctcagtacag agaaatcaat gctggtttac ccacaattgt cactttccct ttcatgtttg
     2281 ccatcatgtt tggtgatatg ggtcacgggt tcttaatgac cttagccgca ttgtctcttg
     2341 tattgaatga aaagaaaatc aacaaaatga aaagaggcga aattttcgat atggccttca
     2401 ctggtagata cattattttg ttgatgggtg tcttttccat gtacacaggt ttcctttaca
     2461 acgatatctt ctctaaaact atgactattt tcaagtctgg ttggaaatgg cctgatcatt
     2521 ggaaaaaagg tgagagtatt actgctacat cggtgggtac ataccctatc ggtttagatt
     2581 gggcttggca tggaactgaa aatgctttgt tattttctaa ttcttacaaa atgaaactat
     2641 caattttaat ggggttcatc cacatgacct attcttattt cttttcgttg gctaaccacc
     2701 tatactttaa ctctatgatt gatatcatcg gtaactttat tcctggtttg ctatttatgc
     2761 aaggtatctt tggttatctt tccgtttgta ttgtttacaa atgggctgtt gattgggtta
     2821 aggacggaaa gcctgctcca ggtttgttaa atatgttgat caacatgttt ttatcaccag
     2881 gaactattga cgatgaatta taccctcatc aagcaaaggt ccaagtgttt ttgttgttga
     2941 tggccttggt ttgtattcct tggttgctat tggtgaagcc attacatttc aaattcactc
     3001 ataaaaagaa atctcacgaa ccactgccat cgactgaagc agatgctagt tctgaagatt
     3061 tggaagcaca acaattaatt tccgcgatgg acgccgatga cgctgaagaa gaagaagttg
     3121 gttctggatc tcatggtgaa gactttggtg atattatgat tcatcaagtt attcatacaa
     3181 ttgaattctg tttgaattgt gtttcgcaca ctgcatccta tttacgttta tgggccttat
     3241 cattggcaca tgctcaattg tctagtgttt tatggacaat gacaattcaa attgcctttg
     3301 gatttagagg atttgtgggt gtgtttatga cggttgcact ttttgccatg tggttcgcac
     3361 taacatgtgc agttcttgtt ttgatggaag gtacatctgc catgcttcat tccttacgtt
     3421 tgcactgggt tgaatctatg tccaagtttt tcgtgggtga aggtttacca tacgaaccat
     3481 tcgcatttga gtataaagac atggaagtcg ctgttgctag tgcaagctct tccgcttcaa
     3541 gcgcttcgta cgctgcaggt cgtgggattc ctgggttaat taacatcgct agcatggtga
     3601 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     3661 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     3721 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     3781 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     3841 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     3901 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     3961 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     4021 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     4081 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     4141 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     4201 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     4261 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtagcctagg tgaggcgcgc
     4321 cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat aagttataaa
     4381 aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt
     4441 cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagtatg aggtcgctct
     4501 tattgaccac acctctaccg gcagatccgc tagggataac agggtaatat ctgcaggtcg
     4561 acaaccctta atataacttc gtataatgta tgctatacga agttattagg tctagagcgg
     4621 ccgcggaacg atcattcact atatatatat caatttatat atacgtatgt gtaattgaag
     4681 aaagatacgt tttttcctct attgagaggc ctgctggatg aatagcttta ccttttctaa
     4741 atccttgata ccatcagtct ctactcctcc acttacatcg acaccgatcg catttggtaa
     4801 catattaatg gcaacagaaa cgttatcagg attcaatcca ccagcgataa tgaattttat
     4861 ctcgggatga cttgcagacc aactggaaat tgcactccaa ttcaatttct caccagtgcc
     4921 accttcacca gaatcgaaca acgtcagcac attgtctacg tgttcataca ggtccagtag
     4981 taattcacaa tcctgtggga actggaacct cttaatgatt ggaattgaag atgggatcaa
     5041 agatctgtat tctttaatat cttcatctcc atgtaattgt atcacatcta gattatattc
     5101 gtggtacagt tgaaggacat catcaacgga ctgatttcta aacaccccga ctagtttagt
     5161 acctttcacg ttctcttgtt ggtgaactgc agttgaaata cctttcgcaa cagatgagtc
     5221 aatggttctt ttcctaccgg gaacacaaat gatacctaag taatcagcac catcatccac
     5281 agcagtcttt gcagcttcaa cggtttgcaa accacacact ttaacgagca tcacagtaga
     5341 tttcgcaaga gggtagctcg ctcagagtac ccaagtaaat gattagtaaa ctgatgtttg
     5401 atagttcaat ttttcaatga aataacctta tattaaaatt gatattacta ttatacaaaa
     5461 ataaagaata aaggatttga gtttatacat aaaataccat tattatttgt tcagtgagag
     5521 atactggggt atatgggatg tgtgtagtga taccatgcaa tcatgtatca aacatgggcc
     5581 cggtaccgag ctctcgagaa cccttaatat aacttcgtat aatgtatgct atacgaagtt
     5641 attaggtgat atcagatcca ctagtggcct atgcggccgc ggatctgccg gtctccctat
     5701 agtgagtcgt attaatttcg ataagccagg ttaacctgca ttaatgaatc ggccaacgcg
     5761 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5821 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5881 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5941 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6001 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6061 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6121 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6181 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6241 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6301 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6361 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6421 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6481 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6541 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6601 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6661 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6721 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6781 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6841 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6901 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6961 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7021 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7081 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7141 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7201 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7261 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7321 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7381 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7441 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7501 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7561 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7621 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7681 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     7741 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     7801 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     7861 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     7921 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgg
     7981 acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac
     8041 gatttaggtg acactataga
//
