LOCUS       sequence-265896-        8062 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of VPH1-mTagBFP2 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 8062)
  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 8062)
  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: ZJOM143
FEATURES             Location/Qualifiers
     source          1..8062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      3893..4080
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3948..3972)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    4132..4165
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5168..5201
                     /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(5254..5273)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5255..5273)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5360..5377)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5531..6119)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5611..5630)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6290..7081,7082..7150))
                     /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     6913..6932
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7151..7255)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7323..7341
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7379..7401)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7501..7520
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7601..7619
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7601..7618
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gttctcagct ttgccatttg cactcggtat cttccctcaa agacaagcaa tccatttaaa
       61 taaactagaa cctgaactac atggcaagaa ggataaggat gggaagccaa ttgaaaaggt
      121 ctattttaac agaggtattt agaagtgaag agacaaattt atatagttat agaataaata
      181 tcagatagat aagaaaaaaa aattagttaa acattaatat atatatatgt gtagtgactg
      241 acatacgtat gactgctagt aatccagttg ccgagctatt gttgcagatt gaaatgtcat
      301 cacgtggaca ttatatcatc attaacgact aatgcgtcaa aaaaaaaatg ggctaaaaaa
      361 aaaaaaagca aaaaaaagaa actcagataa gtcgatatgg aaaacctttt ttcttgagtg
      421 caaccctttt agaggttaca aaacaacaaa agtccaaata agtccatagc ttttgcctct
      481 caatatatct atacttgctt agagggctac ctgtgtgtat ttgctagctt ttacctttct
      541 ctatacaaca agtgagggtt tgaactttta aatccctttt aatttgaatc aaaaaaaaaa
      601 catttaaagg ttacacaagg aaaataatgg cagagaagga ggaagcgatt tttcgctctg
      661 ctgaaatggc tttagtccaa ttctatattc ctcaagaaat ttcaagagac tctgcttaca
      721 ctttaggtca attgggtctt gttcaattcc gtgacttgaa ctctaaggtg cgtgcgtttc
      781 aaagaacttt cgtgaacgaa attagaagac tggataatgt agaaagacag tatcgttact
      841 tttattctct tttgaagaaa cacgatatta agctctacga aggagacacg gacaaatatt
      901 tggacggctc aggtgaattg tacgttccac caagcggttc agtgatagat gattatgtcc
      961 ggaacgcttc atatttggaa gaaagattga ttcaaatgga ggatgcaacc gatcaaattg
     1021 aagtccagaa aaatgacttg gaacagtatc gctttatttt gcagtcaggt gatgaatttt
     1081 tcttgaaggg tgataatacc gacagcactt cctatatgga tgaagacatg atcgacgcta
     1141 atggggaaaa cattgctgct gctatcggtg cttctgtaaa ctatgtcact ggtgtcattg
     1201 ctagagacaa agttgccacc ttagaacaaa ttctttggag agtattaaga ggtaaccttt
     1261 tcttcaaaac tgttgaaatt gaacaacctg tttatgatgt caaaaccagg gagtataaac
     1321 ataaaaatgc ttttatcgta ttttctcacg gtgatctgat tattaaaaga atcagaaaga
     1381 ttgcggaatc attggatgcc aatctttacg atgttgactc ttccaacgag ggtagatcac
     1441 aacaattggc caaggtcaac aagaatttga gtgatttgta cacagttttg aaaaccactt
     1501 ctaccacttt agaaagtgaa ttatatgcca ttgccaaaga attggactct tggttccaag
     1561 atgttacccg tgaaaaggcg atttttgaaa ttttgaacaa gtctaactat gataccaata
     1621 gaaagatttt gattgctgaa ggttggatac caagagacga attggctact ttgcaagctc
     1681 gtcttggtga aatgatcgca agattgggta ttgatgtccc atccattatc caagtcctgg
     1741 atacaaacca cactccacct accttccaca gaactaacaa gtttactgct ggtttccaaa
     1801 gtatctgtga ctgttacggt attgctcagt acagagaaat caatgctggt ttacccacaa
     1861 ttgtcacttt ccctttcatg tttgccatca tgtttggtga tatgggtcac gggttcttaa
     1921 tgaccttagc cgcattgtct cttgtattga atgaaaagaa aatcaacaaa atgaaaagag
     1981 gcgaaatttt cgatatggcc ttcactggta gatacattat tttgttgatg ggtgtctttt
     2041 ccatgtacac aggtttcctt tacaacgata tcttctctaa aactatgact attttcaagt
     2101 ctggttggaa atggcctgat cattggaaaa aaggtgagag tattactgct acatcggtgg
     2161 gtacataccc tatcggttta gattgggctt ggcatggaac tgaaaatgct ttgttatttt
     2221 ctaattctta caaaatgaaa ctatcaattt taatggggtt catccacatg acctattctt
     2281 atttcttttc gttggctaac cacctatact ttaactctat gattgatatc atcggtaact
     2341 ttattcctgg tttgctattt atgcaaggta tctttggtta tctttccgtt tgtattgttt
     2401 acaaatgggc tgttgattgg gttaaggacg gaaagcctgc tccaggtttg ttaaatatgt
     2461 tgatcaacat gtttttatca ccaggaacta ttgacgatga attataccct catcaagcaa
     2521 aggtccaagt gtttttgttg ttgatggcct tggtttgtat tccttggttg ctattggtga
     2581 agccattaca tttcaaattc actcataaaa agaaatctca cgaaccactg ccatcgactg
     2641 aagcagatgc tagttctgaa gatttggaag cacaacaatt aatttccgcg atggacgccg
     2701 atgacgctga agaagaagaa gttggttctg gatctcatgg tgaagacttt ggtgatatta
     2761 tgattcatca agttattcat acaattgaat tctgtttgaa ttgtgtttcg cacactgcat
     2821 cctatttacg tttatgggcc ttatcattgg cacatgctca attgtctagt gttttatgga
     2881 caatgacaat tcaaattgcc tttggattta gaggatttgt gggtgtgttt atgacggttg
     2941 cactttttgc catgtggttc gcactaacat gtgcagttct tgttttgatg gaaggtacat
     3001 ctgccatgct tcattcctta cgtttgcact gggttgaatc tatgtccaag tttttcgtgg
     3061 gtgaaggttt accatacgaa ccattcgcat ttgagtataa agacatggaa gtcgctgttg
     3121 ctagtgcaag ctcttccgct tcaagccttt ctcttgaagt tgctgaagaa atgagcgaac
     3181 tgatcaaaga gaacatgcac atgaagctgt acatggaagg caccgttgac aaccaccact
     3241 ttaagtgcac ctctgagggt gagggtaagc cctacgaagg cacccaaacc atgcgtatca
     3301 aagttgtgga gggcggtcca ctgccattcg cttttgacat tctggccacc agcttcctgt
     3361 acggttccaa aaccttcatt aaccatactc agggcattcc tgatttcttc aaacagagct
     3421 ttccggaagg tttcacctgg gagcgtgtca ccacgtatga agatggtggt gtgttgaccg
     3481 ccacccaaga tacctccctg caagatggct gtctgatcta taacgtgaaa attcgtggcg
     3541 tcaactttac tagcaatggt cccgtgatgc agaagaaaac cctgggttgg gaggccttta
     3601 cagaaaccct gtatcctgcc gatggtggcc tggagggccg taacgacatg gcactgaagc
     3661 tggttggtgg cagccatttg atcgcaaatg ccaagaccac ttacagaagc aagaaacccg
     3721 caaaaaatct gaagatgccg ggtgtttact atgtcgacta ccgtctggaa agaattaaag
     3781 aagcaaataa tgagacttac gtggagcagc acgaggttgc agtcgcgaga tattgcgact
     3841 tgcctagcaa gctgggtcat aaactgaatt gaggcgcgcc acttctaaat aagcgaattt
     3901 cttatgattt atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt
     3961 ttaaagtgac tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta
     4021 ggtcaggttg ctttctcagg tatagtatga ggtcgctctt attgaccaca cctctaccgg
     4081 cagatccgct agggataaca gggtaatatc tgcaggtcga caacccttaa tataacttcg
     4141 tataatgtat gctatacgaa gttattaggt ctagagcggc cgcggaacga tcattcacta
     4201 tatatatatc aatttatata tacgtatgtg taattgaaga aagatacgtt ttttcctcta
     4261 ttgagaggcc tgctggatga atagctttac cttttctaaa tccttgatac catcagtctc
     4321 tactcctcca cttacatcga caccgatcgc atttggtaac atattaatgg caacagaaac
     4381 gttatcagga ttcaatccac cagcgataat gaattttatc tcgggatgac ttgcagacca
     4441 actggaaatt gcactccaat tcaatttctc accagtgcca ccttcaccag aatcgaacaa
     4501 cgtcagcaca ttgtctacgt gttcatacag gtccagtagt aattcacaat cctgtgggaa
     4561 ctggaacctc ttaatgattg gaattgaaga tgggatcaaa gatctgtatt ctttaatatc
     4621 ttcatctcca tgtaattgta tcacatctag attatattcg tggtacagtt gaaggacatc
     4681 atcaacggac tgatttctaa acaccccgac tagtttagta cctttcacgt tctcttgttg
     4741 gtgaactgca gttgaaatac ctttcgcaac agatgagtca atggttcttt tcctaccggg
     4801 aacacaaatg atacctaagt aatcagcacc atcatccaca gcagtctttg cagcttcaac
     4861 ggtttgcaaa ccacacactt taacgagcat cacagtagat ttcgcaagag ggtagctcgc
     4921 tcagagtacc caagtaaatg attagtaaac tgatgtttga tagttcaatt tttcaatgaa
     4981 ataaccttat attaaaattg atattactat tatacaaaaa taaagaataa aggatttgag
     5041 tttatacata aaataccatt attatttgtt cagtgagaga tactggggta tatgggatgt
     5101 gtgtagtgat accatgcaat catgtatcaa acatgggccc ggtaccgagc tctcgagaac
     5161 ccttaatata acttcgtata atgtatgcta tacgaagtta ttaggtgata tcagatccac
     5221 tagtggccta tgcggccgcg gatctgccgg tctccctata gtgagtcgta ttaatttcga
     5281 taagccaggt taacctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5341 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5401 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5461 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5521 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5581 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5641 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5701 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5761 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5821 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5881 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5941 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6001 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6061 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6121 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6181 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6241 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6301 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6361 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6421 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6481 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6541 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6601 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6661 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6721 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6781 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6841 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6901 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6961 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7021 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7081 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7141 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7201 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7261 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     7321 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     7381 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     7441 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     7501 cggcatcaga gcagattgta ctgagagtgc accatatgga catattgtcg ttagaacgcg
     7561 gctacaatta atacataacc ttatgtatca tacacatacg atttaggtga cactatagaa
     7621 cgcggccgcc agctgaagct tcgtacgctg caggtcgtgg gattcctggg ttaattaaca
     7681 tcgctagctg tggtgttgcg ttaggtctaa ataacctagt tcccaggttg aaaaacattt
     7741 ctccacattc aaagttgatc ttgggccgtt tggtgccatt tgccgccgtg gtcagtgccg
     7801 gtattgtcaa cgtcttccta atgagaggca acgaaattag aaagggtatt tctgtgtttg
     7861 actctaacgg tgacgaagtc ggaaagtcca agaaggctgc cttcatggca gttggtgaaa
     7921 ctgcattaag tagagtgatt aacgccactc caactatggt catcccacct ttgattttgg
     7981 ttagactaca acgcggtgtc ctcaaaggta agtctctcgg tgtgcaaact ttggccaatt
     8041 tggggttgat ctcagtcaca at
//
