LOCUS       sequence-308131-        7605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of VPS4-DuDre as an additional copy in genome, use 
            auxotrophic marker TRP1(Kluyveromyces lactis). Marker for yeast late
            endosome..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7605)
  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 7605)
  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: ZJOM43
FEATURES             Location/Qualifiers
     source          1..7605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      3881..4068
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3936..3960)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    4120..4153
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5156..5189
                     /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(5242..5261)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5243..5261)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5348..5365)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5519..6107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5599..5618)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6278..7069,7070..7138))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVARPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6901..6920
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7139..7243)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7311..7329
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7367..7389)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7489..7508
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(7589..7605,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(7589..7605,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcatcac ttttgttcct tttgtcgttt ttcttctttt
       61 agattccatc ttgaaaagag atgaaatgga caaagtaaca taatttagac ccaacaaagt
      121 aaaaagaagg ttgggttggc attgaagatc acaatgggaa ttcaggaaaa gacattgggg
      181 atacgaaaag agcgcaaatt ggtggttgtg cctcgcgaaa gaaatcatgt tagacacgct
      241 tcacaaagga cacgttctaa aaactacaag aatatctcca aaaagagggc tcaacaacac
      301 gcgtttggat tcaatattgc taagactctt gccaagatcc aagcgtttgt atgggggtct
      361 cccgcagatg aggaagaaga aagcgttgtc ccattgtcta aaaattctca ggattgtgtg
      421 cctctacaat ggcaagcaaa gtttgcgcaa ttgcggcagc agctgcactc tacacaaaag
      481 gaactacaat ttgtcaagga aaagtgtcat ctgttgcagt cggtgttgga cgacgcaaac
      541 atcgatcaaa gatacctgga gtctcgtcga gacatgaaaa atattgagcg agacaacctc
      601 aaaccgacag aaaatttacc accttcgcca gtaagagcag taaacccgtt agtgacttca
      661 agtccgattc acatgtcgcc actccagtcg cgccaacgac cagtttcttc cctacagccc
      721 ccaaagggcc ccaacttcta cgccaagtat cctaagcttc ctcaaacgaa cattctcaga
      781 gaatctccga ctgaggattc ggttccgcac gctgaataga aacacaacac gtcatggttg
      841 ttctttctgt acatctattt attcttgctt gatgatattt attgtattgc tttacgggta
      901 cccccgctgg gtgaaaaaaa aaaaaaaaat aagaacttca tgatatcgaa cgatttaatc
      961 gtgggttgag gacatggaag acaaaaataa agcagcatag agtgcctata gtagatgggg
     1021 tacaaatgag cacgggagat tttttaacta agggaataga actggttcaa aaggccattg
     1081 acctggacac agccacgcag tatgaggagg cgtatacagc atactataac ggattagatt
     1141 atttgatgtt ggctttgaaa tacgagaaaa accctaagtc gaaggatttg ataagagcga
     1201 agtttacaga gtatctaaac cgtgcagagc agttgaaaaa gcacttggaa agtgaagagg
     1261 caaacgcggc aaagaaatct cctagcgctg gtagtggttc caacggtggt aataagaaaa
     1321 tctctcagga agagggtgag gataatggtg gagaagacaa caaaaagcta agaggtgcct
     1381 tatcgagtgc tattttgtct gagaagccca atgttaagtg ggaagacgtg gctggactgg
     1441 aaggagccaa agaggccctt aaagaagccg ttattttacc tgtcaagttc ccacatttgt
     1501 tcaaaggtaa ccgtaagccc acttcgggaa tcttattgta cggaccacca ggtacaggta
     1561 aatcatattt ggcaaaagca gtggccaccg aggccaattc tacctttttc agtgttagtt
     1621 ccagtgattt ggtttctaag tggatgggtg aatctgaaaa acttgtcaaa cagttatttg
     1681 cgatggcaag agaaaataaa ccttctatta tttttataga tgaagtggat gcgctaacag
     1741 gtaccagagg agaaggagag agcgaagcaa gtagaagaat taaaacagaa ttattggtac
     1801 aaatgaatgg tgttgggaat gactcccaag gtgttttggt ccttggcgca acaaatatcc
     1861 catggcaact ggacagtgcc atcaggagaa gatttgaaag aagaatatat ataccattac
     1921 cagatttggc ggcaagaacc accatgttcg agattaacgt cggcgacact ccatgcgtat
     1981 tgaccaagga agattataga actttaggtg caatgactga aggttattcc gggagtgata
     2041 ttgctgtagt ggtgaaagat gcgctaatgc aaccgataag aaaaattcaa agcgcgacac
     2101 atttcaaaga tgtctctacg gaggacgacg aaacaagaaa actaacacca tgctctccag
     2161 gtgatgatgg cgccatcgag atgagttgga cggatattga agctgatgaa ttaaaagaac
     2221 cagatctcac cataaaggat ttcttaaagg ctatcaaatc gacgagacct accgtgaatg
     2281 aggatgactt gctgaagcaa gaacagttca ctagagattt tggtcaagaa ggtaacgctt
     2341 cgtacgctgc aggtcgtggg attcctgggt taattaacat cgctagcatg gtttccaaag
     2401 gtgaagagaa caatatggct gtcatcaagc ccttcatgag attcaaggtg cacatggagg
     2461 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggcaag ccctacgagg
     2521 gcacccagac cgccaagctg caggtgacca agggcggccc cctgcccttc gcctgggaca
     2581 tcctgtcccc ccagttccag tacggctcca aggtgtacgt gaagcacccc gccgacatcc
     2641 ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg ggagagagtg atgaacttcg
     2701 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc accttcatct
     2761 accacgtgaa gttcatcggc gtgaacttcc cctccgacgg ccccgtaatg cagaagaaga
     2821 ctctgggctg ggagccctcc accgagagac tgtaccccag agacggcgtg ctgaagggcg
     2881 agatccacaa ggcgctgaag ctgaagggcg gcggccacta cctggtggag ttcaagtcaa
     2941 tctacatggc caagaagccc gtgaagctgc ccggctacta ctacgtggac tccaagctgg
     3001 acatcacctc ccacaacgag gactacaccg tggtggagca gtacgagaga gccgaggcca
     3061 gacaccacct gttcttaatt ccagttccaa cgcgtgatct tgagaagcca ttcaacgtca
     3121 tcaagccctt catgagattc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
     3181 agatcgaggg cgagggcgag ggcaagccct acgagggcac ccagaccgcc aagctgcagg
     3241 tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag ttccagtacg
     3301 gctccaaggt gtacgtgaag caccccgccg acatccccga ctacaagaag ctgtccttcc
     3361 ccgagggctt caagtgggag agagtgatga acttcgagga cggcggcgtg gtgaccgtga
     3421 cccaggactc ctccctgcag gacggcacct tcatctacca cgtgaagttc atcggcgtga
     3481 acttcccctc cgacggcccc gtaatgcaga agaagactct gggctgggag ccctccaccg
     3541 agagactgta ccccagagac ggcgtgctga agggcgagat ccacaaggcg ctgaagctga
     3601 agggcggcgg ccactacctg gtggagttca agtcaatcta catggccaag aagcccgtga
     3661 agctgcccgg ctactactac gtggactcca agctggacat cacctcccac aacgaggact
     3721 acaccgtggt ggagcagtac gagagagccg aggccagaca ccacctgttc ttaattgtcg
     3781 ttctgaacag agctgagact ccattgccat tggatccaac cggtaaggtc aaggctgagc
     3841 tggatactag gcctaggtga ggcgcgccac ttctaaataa gcgaatttct tatgatttat
     3901 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     3961 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     4021 ttctcaggta tagtatgagg tcgctcttat tgaccacacc tctaccggca gatccgctag
     4081 ggataacagg gtaatatctg caggtcgaca acccttaata taacttcgta taatgtatgc
     4141 tatacgaagt tattaggtct agagcggccg cggaacgatc attcactata tatatatcaa
     4201 tttatatata cgtatgtgta attgaagaaa gatacgtttt ttcctctatt gagaggcctg
     4261 ctggatgaat agctttacct tttctaaatc cttgatacca tcagtctcta ctcctccact
     4321 tacatcgaca ccgatcgcat ttggtaacat attaatggca acagaaacgt tatcaggatt
     4381 caatccacca gcgataatga attttatctc gggatgactt gcagaccaac tggaaattgc
     4441 actccaattc aatttctcac cagtgccacc ttcaccagaa tcgaacaacg tcagcacatt
     4501 gtctacgtgt tcatacaggt ccagtagtaa ttcacaatcc tgtgggaact ggaacctctt
     4561 aatgattgga attgaagatg ggatcaaaga tctgtattct ttaatatctt catctccatg
     4621 taattgtatc acatctagat tatattcgtg gtacagttga aggacatcat caacggactg
     4681 atttctaaac accccgacta gtttagtacc tttcacgttc tcttgttggt gaactgcagt
     4741 tgaaatacct ttcgcaacag atgagtcaat ggttcttttc ctaccgggaa cacaaatgat
     4801 acctaagtaa tcagcaccat catccacagc agtctttgca gcttcaacgg tttgcaaacc
     4861 acacacttta acgagcatca cagtagattt cgcaagaggg tagctcgctc agagtaccca
     4921 agtaaatgat tagtaaactg atgtttgata gttcaatttt tcaatgaaat aaccttatat
     4981 taaaattgat attactatta tacaaaaata aagaataaag gatttgagtt tatacataaa
     5041 ataccattat tatttgttca gtgagagata ctggggtata tgggatgtgt gtagtgatac
     5101 catgcaatca tgtatcaaac atgggcccgg taccgagctc tcgagaaccc ttaatataac
     5161 ttcgtataat gtatgctata cgaagttatt aggtgatatc agatccacta gtggcctatg
     5221 cggccgcgga tctgccggtc tccctatagt gagtcgtatt aatttcgata agccaggtta
     5281 acctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5341 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5401 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5461 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5521 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5581 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5641 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5701 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5761 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5821 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5881 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5941 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6001 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6061 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6121 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6181 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6241 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6301 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6361 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6421 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6481 gcagaagtgg tcgtgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6541 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6601 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6661 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6721 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6781 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6841 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6901 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6961 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7021 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7081 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7141 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7201 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7261 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     7321 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     7381 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     7441 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     7501 agattgtact gagagtgcac catatggaca tattgtcgtt agaacgcggc tacaattaat
     7561 acataacctt atgtatcata cacatacgat ttaggtgaca ctata
//
