LOCUS       sequence-292111-        6214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of mTagBFP2-Tlg1 as an additional copy in genome, use 
            auxotrophic marker TRP1(Kluyveromyces lactis). Marker for yeast late
            golgi/early endosome..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6214)
  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 6214)
  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: ZJOM108
FEATURES             Location/Qualifiers
     source          1..6214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      2507..2694
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2562..2586)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    2746..2779
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3782..3815
                     /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(3868..3887)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3869..3887)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3974..3991)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4145..4733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4225..4244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4904..5695,5696..5764))
                     /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     5527..5546
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5765..5869)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5937..5955
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5993..6015)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6115..6134
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaggtga cactatagaa cgcggccgcc agctgaagct tcgtacgctg caggtcgtgg
       61 gattcctggg ttaattaaca tcgctagcga tagtccccca ttttttttta tgttcgccca
      121 tggtattcca gtggcccttt ttctccacaa tttttgtatc caattttcct ccaccacttg
      181 tctatatcca aaggtttaat agtaaggaaa aataaataca catgtatatt tatataatta
      241 tggttatcat ttactagcat tactctcttt ttctccactc ttcagtatat tgctctgctc
      301 gattagatat tcacctaaaa cacgtaacgg atcttctggt tgttgtacgg ctatgagcct
      361 catgcccgct aataaatgcg gtgtcacatt tgtattcagg tattttctag tttgtgaacc
      421 acctaccgtc gcggcaaggt tcacattttc taattttgat attttaggtt cagatgactc
      481 ttcaatgact gaatttgtgc tactcgattc ttctattttt acattttggt tggtgacact
      541 tttgatgttt ttcaaatctt tcgttgcaga agatggtaca ctgtcaaatt gtaagttatc
      601 agagttatta ttgtgattgc cattcatatc gaagcattca ttgtcatgtt tatcgtagtt
      661 cttattgtct tcgcgttgaa tttgctccat tggaatatca gcattacttg aagtatcctc
      721 taccatagga accgtaactt cattatgttg gggactattc tcactttcat tcattgtata
      781 ggtttcccta attctcttaa gagaacctgc ttatgaaggc ttttttttgt ttggatcacc
      841 atctgttttt ctttttttat cgagttttta attatccacg agatattcga acctacgaca
      901 taaagtagta aatatcagaa gataggacaa ttcgaagcac acaacacaga acatatacta
      961 aaattatatt tgctgtcttt ctgacaaggt ttgtgaacat tttaatttct acgttttccc
     1021 tgtaactgaa gtaaacgtga aagacgttta actgtaaaat tgtattgcta agatcctttc
     1081 tttaacaaat gagcgaactg atcaaagaga acatgcacat gaagctgtac atggaaggca
     1141 ccgttgacaa ccaccacttt aagtgcacct ctgagggtga gggtaagccc tacgaaggca
     1201 cccaaaccat gcgtatcaaa gttgtggagg gcggtccact gccattcgct tttgacattc
     1261 tggccaccag cttcctgtac ggttccaaaa ccttcattaa ccatactcag ggcattcctg
     1321 atttcttcaa acagagcttt ccggaaggtt tcacctggga gcgtgtcacc acgtatgaag
     1381 atggtggtgt gttgaccgcc acccaagata cctccctgca agatggctgt ctgatctata
     1441 acgtgaaaat tcgtggcgtc aactttacta gcaatggtcc cgtgatgcag aagaaaaccc
     1501 tgggttggga ggcctttaca gaaaccctgt atcctgccga tggtggcctg gagggccgta
     1561 acgacatggc actgaagctg gttggtggca gccatttgat cgcaaatgcc aagaccactt
     1621 acagaagcaa gaaacccgca aaaaatctga agatgccggg tgtttactat gtcgactacc
     1681 gtctggaaag aattaaagaa gcaaataatg agacttacgt ggagcagcac gaggttgcag
     1741 tcgcgagata ttgcgacttg cctagcaagc tgggtcataa actgaatctt tctcttgaag
     1801 ttgctgaaga aatgaacaac agtgaagatc cgtttcaaca agttgttaag gacaccaagg
     1861 agcaattgaa ccgcataaac aattacataa ctcgtcacaa tactgctggt gatgacgatc
     1921 aagaggagga aatacaagat attttaaagg atgttgagga aacaatagtt gatttggaca
     1981 gaagcataat cgtaatgaaa agggatgaaa acgaagacgt gagtggtagg gaagcacaag
     2041 ttaaaaatat aaaacagcaa cttgatgctc tgaagttgcg ttttgatcga agaatacagg
     2101 aatctactca aacaactatt cctctagaag agacggtgga aaattcaaca cttaacacca
     2161 gcatggctga gaacaatgat ggtggtatgt ccaatccgtt tcaggaacaa atgttaagag
     2221 aacaagatgt tcatttagat ggtattcaca agacaatgca aaatttgcat attcaagctc
     2281 aaacaatggg ggatgaatta gagaaccagg gacaattgtt ggataatatg gacgagggta
     2341 tggacggtgt tgtaaataag ctggctagag gtcgtaggca attggaatgg gtctacgaaa
     2401 aaaataaaga aaaatacgac gattgttgta taggacttct tattgtcgtc ttgatagttt
     2461 tattagtttt ggcattcatt gcttgaggcg cgccacttct aaataagcga atttcttatg
     2521 atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag
     2581 tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag
     2641 gttgctttct caggtatagt atgaggtcgc tcttattgac cacacctcta ccggcagatc
     2701 cgctagggat aacagggtaa tatctgcagg tcgacaaccc ttaatataac ttcgtataat
     2761 gtatgctata cgaagttatt aggtctagag cggccgcgga acgatcattc actatatata
     2821 tatcaattta tatatacgta tgtgtaattg aagaaagata cgttttttcc tctattgaga
     2881 ggcctgctgg atgaatagct ttaccttttc taaatccttg ataccatcag tctctactcc
     2941 tccacttaca tcgacaccga tcgcatttgg taacatatta atggcaacag aaacgttatc
     3001 aggattcaat ccaccagcga taatgaattt tatctcggga tgacttgcag accaactgga
     3061 aattgcactc caattcaatt tctcaccagt gccaccttca ccagaatcga acaacgtcag
     3121 cacattgtct acgtgttcat acaggtccag tagtaattca caatcctgtg ggaactggaa
     3181 cctcttaatg attggaattg aagatgggat caaagatctg tattctttaa tatcttcatc
     3241 tccatgtaat tgtatcacat ctagattata ttcgtggtac agttgaagga catcatcaac
     3301 ggactgattt ctaaacaccc cgactagttt agtacctttc acgttctctt gttggtgaac
     3361 tgcagttgaa atacctttcg caacagatga gtcaatggtt cttttcctac cgggaacaca
     3421 aatgatacct aagtaatcag caccatcatc cacagcagtc tttgcagctt caacggtttg
     3481 caaaccacac actttaacga gcatcacagt agatttcgca agagggtagc tcgctcagag
     3541 tacccaagta aatgattagt aaactgatgt ttgatagttc aatttttcaa tgaaataacc
     3601 ttatattaaa attgatatta ctattataca aaaataaaga ataaaggatt tgagtttata
     3661 cataaaatac cattattatt tgttcagtga gagatactgg ggtatatggg atgtgtgtag
     3721 tgataccatg caatcatgta tcaaacatgg gcccggtacc gagctctcga gaacccttaa
     3781 tataacttcg tataatgtat gctatacgaa gttattaggt gatatcagat ccactagtgg
     3841 cctatgcggc cgcggatctg ccggtctccc tatagtgagt cgtattaatt tcgataagcc
     3901 aggttaacct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3961 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4021 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4081 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4141 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4201 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4261 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4321 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4381 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4441 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4501 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4561 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4621 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4681 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4741 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4801 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4861 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4921 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4981 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5041 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5101 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5161 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5221 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5281 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5341 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5401 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5461 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5521 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5581 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5641 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5701 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5761 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5821 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5881 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     5941 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     6001 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     6061 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     6121 cagagcagat tgtactgaga gtgcaccata tggacatatt gtcgttagaa cgcggctaca
     6181 attaatacat aaccttatgt atcatacaca tacg
//
