LOCUS       sequence-256879-        8464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of SEC26-mTagBFP2 as an additional copy in genome, use 
            auxotrophic marker TRP1(Kluyveromyces lactis). Marker for yeast 
            early golgi..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8464)
  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 8464)
  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: ZJOM96
FEATURES             Location/Qualifiers
     source          1..8464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      4737..4924
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4792..4816)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    4976..5009
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    6015..6048
                     /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(6101..6120)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6102..6120)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6207..6224)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6378..6966)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6458..6477)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7137..7928,7929..7997))
                     /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     7760..7779
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7998..8102)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8170..8188
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8226..8248)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8348..8367
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(8448..8464,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(8448..8464,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg tgggattcct gggttaatta
       61 acatcgctag cttcaccgtt aacatctgtc gtatcgaaca aatcggaaaa gtccgtgcct
      121 ttggtttcta gttacgtaga agaactgcga ctaaagtatt acaagacttc taacttttta
      181 caagcgcctc ctaacttgcc ggttgctcta aaacaaaaaa ataatttgat tcaacccaag
      241 aatattaagg tcaagttgag gacaagttct aagcaaattg gtatcaagca tggtaaagtt
      301 aaacagaaat tactggcgtt ggagacaaga aatgaagaat cagatggaac cgctacaggg
      361 ctgaaaaata aaatcaatgt tgaccacaca aaggaattcc ataaattgtt agggaaggaa
      421 aatgaaactg gctctatttc aaaaaaagaa ggcactgacg cagaacaagc cgaagactac
      481 ctcaaggaca tacccggtga tgaagcatat aacagtgatg atataatggc gcctttacga
      541 gaaaaaagag ggcaaaacgg ttctgttgat tccgtgtcaa gatctaacac ggtagtcagc
      601 tactacacaa ggtctcagaa cagaatgaga agtggaacac tggataatga ttacgtgaac
      661 agacaaaagc ttcctacaca tatctctctt caagattatc gtgatgctaa tgctagaagt
      721 aatatatcgc gtcaggactc tgtctccaca acgaactctg atgtggtaga cctcagctat
      781 tctctggggc atggcttgcg tgtggcaaac cctgattcag acccagaatg atatccttct
      841 ctgtagtttt gtagatgtca tatatgtacg tttatggaac agcattttag aaaagtatta
      901 cccagcttat cacttcgttt tttttttctt tccgtgtact cgctacacgt agaaagaaat
      961 cagaaaacaa cagctcgaca agtgaaattt gacgttcatt aagactcagt taagattgcc
     1021 ttgagaataa acaaaagtaa tcacagttaa ctattgaaca agagtgcacc tatgacttca
     1081 ctttcttcac agccagcgta cacgttggtt ttcgatcctt ctccgagtat ggagacttac
     1141 tcgagtaccg attttcagaa agctcttgaa aagggatctg atgaacaaaa aattgacacg
     1201 atgaaatcaa ttttagttac aatgctggaa ggaaatccaa tgcctgaatt gttgatgcac
     1261 ataataagat ttgtcatgcc ttctaaaaat aaggaattaa aaaagctttt gtacttctac
     1321 tgggaaattg ttcccaaact agctgaagat ggaaaattga gacatgaaat gattcttgtc
     1381 tgtaatgcca ttcaacacga tttgcaacat cctaatgaat atattagagg taacacatta
     1441 aggtttttaa cgaaattgag agaggccgaa ctcttagaac agatggttcc ctctgtctta
     1501 gcgtgcttgg aataccgtca tgcatatgtt cgtaagtatg caatcctagc agttttctcc
     1561 attttcaagg tcagcgaaca tttacttccc gatgctaaag aaatcatcaa ttcgttcata
     1621 gtagctgaaa ctgatccaat atgtaaaaga aatgcattta ttgggttagc tgaattagat
     1681 cgtgaaaatg ccttacacta tttagagaac aatattgctg atatagaaaa cctagaccct
     1741 ttattacaag ctgtctttgt tcaatttatc agacaagatg caaacaggac ccctgctttg
     1801 aaagcccaat atatcgaatt attgatggaa ctgctttcga ccacgacttc cgatgaagtc
     1861 atcttcgaga ccgcattagc cctaactgtg ttgtctgcca atccaaatgt cttggttcct
     1921 gcggttaaca aattgattga cttggccgtc aaggtttctg ataataacat taagttaatt
     1981 gttctagacc gtattcaaga catcaatgct aataacgtag gtgctttgga agagttaacc
     2041 ctggatattt tgagagtctt gaatgcagaa gatttagacg ttcgttcaaa ggcgcttgat
     2101 atttcaatgg acttggccac atccagaaat gctgaagatg ttgttcagct tttgaagaaa
     2161 gagctgcaaa caaccgtaaa taacccagat caagacaagg caatgcagta cagacaattg
     2221 ttaataaaaa ctattcgtac cgtggctgta aactttgtag aaatggcagc aagtgttgtt
     2281 tcgctattat tagatttcat cggtgattta aactcggttg ccgccagtgg tatcattgcc
     2341 tttatcaaag aagtgatcga aaaataccca caacttagag ccaatatcct tgaaaacatg
     2401 gttcaaacgc tagacaaagt gagatctgct aaagcttacc gcggtgcatt atggattatg
     2461 ggtgagtatg ctgaaggaga aagtgagata caacattgtt ggaagcacat tcgtaacagc
     2521 gtaggtgaag ttcctatcct tcaatcagaa atcaaaaagt taacacaaaa ccaagaacac
     2581 accgaagaaa atgaggttga cgctaccgcc aagccaactg gtccagttat tctaccagac
     2641 ggtacgtatg ccactgaaag cgctttcgat gtgaagactt ctcaaaagtc agttaccgat
     2701 gaagaacgtg attctagacc tccaattcgc cggtttgttt taagtggtga tttctacaca
     2761 gctgccattc tggccaacac catcattaaa cttgttttaa aattcgaaaa cgtttccaag
     2821 aacaaaactg tcatcaatgc tctaaaggcg gaagctttac taattttagt tagtattgta
     2881 agagtgggtc aaagctcttt ggtggagaaa aaaattgatg aagattcttt agagagagtt
     2941 atgacatcta tttctatttt attggatgaa gttaatcctg aggaaaagaa ggaagaagtt
     3001 aaacttctgg aggttgcatt cctggacacc accaaatcct cattcaagag acaaattgaa
     3061 attgcaaaga agaacaagca taagagagca ttaaaagaca gttgcaaaaa catcgaacca
     3121 attgatacgc cgatttcttt caggcaattt gctggtgtgg attctactaa tgtgcaaaaa
     3181 gatagtattg aagaagattt acaactggca atgaaaggag atgcaatcca cgctactagc
     3241 agctctagta tttcgaagct gaagaagata gtacctttat gtggcttttc tgatccagtt
     3301 tacgccgagg cttgtattac aaacaatcaa tttgacgtcg tattagatgt tcttcttgtt
     3361 aatcaaacga aagaaacatt gaaaaaccta catgtgcaat ttgcaactct tggtgatttg
     3421 aagattattg acacaccaca gaagaccaac gtgattcctc atggcttcca caaattcact
     3481 gttactgtca aagtttcctc tgctgacaca ggtgtcattt tcggtaatat tatttatgat
     3541 ggtgcgcatg gtgaagatgc tcgttatgtt attttaaacg acgttcatgt tgacattatg
     3601 gattatatca aaccagccac tgctgacgat gaacatttcc gtaccatgtg gaatgcattt
     3661 gagtgggaga acaaaatatc ggtcaaatca caactaccaa cattgcatgc ttatttgaga
     3721 gaactggtca agggaactaa tatgggtatt ctaacaccat cagagtcgtt gggagaagat
     3781 gattgtaggt tcttaagttg taatctgtat gcgaagtcgt cctttggtga agatgcccta
     3841 gccaacttgt gtatcgaaaa ggattccaaa accaatgatg tcataggtta tgttcgtatc
     3901 cgatcaaagg gacaaggttt ggctctgtcc ctaggtgaca gagtggcatt gattgctaag
     3961 aagaccaata aacttgctct cactcatgtt ctttctcttg aagttgctga agaaatgagc
     4021 gaactgatca aagagaacat gcacatgaag ctgtacatgg aaggcaccgt tgacaaccac
     4081 cactttaagt gcacctctga gggtgagggt aagccctacg aaggcaccca aaccatgcgt
     4141 atcaaagttg tggagggcgg tccactgcca ttcgcttttg acattctggc caccagcttc
     4201 ctgtacggtt ccaaaacctt cattaaccat actcagggca ttcctgattt cttcaaacag
     4261 agctttccgg aaggtttcac ctgggagcgt gtcaccacgt atgaagatgg tggtgtgttg
     4321 accgccaccc aagatacctc cctgcaagat ggctgtctga tctataacgt gaaaattcgt
     4381 ggcgtcaact ttactagcaa tggtcccgtg atgcagaaga aaaccctggg ttgggaggcc
     4441 tttacagaaa ccctgtatcc tgccgatggt ggcctggagg gccgtaacga catggcactg
     4501 aagctggttg gtggcagcca tttgatcgca aatgccaaga ccacttacag aagcaagaaa
     4561 cccgcaaaaa atctgaagat gccgggtgtt tactatgtcg actaccgtct ggaaagaatt
     4621 aaagaagcaa ataatgagac ttacgtggag cagcacgagg ttgcagtcgc gagatattgc
     4681 gacttgccta gcaagctggg tcataaactg aattgaggcg cgccacttct aaataagcga
     4741 atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca
     4801 aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc
     4861 tgtaggtcag gttgctttct caggtatagt atgaggtcgc tcttattgac cacacctcta
     4921 ccggcagatc cgctagggat aacagggtaa tatctgcagg tcgacaaccc ttaatataac
     4981 ttcgtataat gtatgctata cgaagttatt aggtctagag cggccgcgga acgatcattc
     5041 actatatata tatcaattta tatatacgta tgtgtaattg aagaaagata cgttttttcc
     5101 tctattgaga ggcctgctgg atgaatagct ttaccttttc taaatccttg ataccatcag
     5161 tctctactcc tccacttaca tcgacaccga tcgcatttgg taacatatta atggcaacag
     5221 aaacgttatc aggattcaat ccaccagcga taatgaattt tatctcggga tgacttgcag
     5281 accaactgga aattgcactc caattcaatt tctcaccagt gccaccttca ccagaatcga
     5341 acaacgtcag cacattgtct acgtgttcat acaggtccag tagtaattca caatcctgtg
     5401 ggaactggaa cctcttaatg attggaattg aagatgggat caaagatctg tattctttaa
     5461 tatcttcatc tccatgtaat tgtatcacat ctagattata ttcgtggtac agttgaagga
     5521 catcatcaac ggactgattt ctaaacaccc cgactagttt agtacctttc acgttctctt
     5581 gttggtgaac tgcagttgaa atacctttcg caacagatga gtcaatggtt cttttcctac
     5641 cgggaacaca aatgatacct aagtaatcag caccatcatc cacagcagtc tttgcagctt
     5701 caacggtttg caaaccacac actttaacga gcatcacagt agatttcgca agagggtagc
     5761 tcgctcagag tacccaagta aatgattagt aaactgatgt ttgatagttc aatttttcaa
     5821 tgaaataacc ttatattaaa attgatatta ctattataca aaaataaaga ataaaggatt
     5881 tgagtttata cataaaatac cattattatt tgttcagtga gagataccgg ggtatatggg
     5941 atgtgtgtag tgataccatg caatcatgta tcaaacatgg gcccggtacc gagctctggt
     6001 cgagaaccct taatataact tcgtataatg tatgctatac gaagttatta ggtgatatca
     6061 gatccactag tggcctatgc ggccgcggat ctgccggtct ccctatagtg agtcgtatta
     6121 atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6181 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6241 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6301 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6361 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6421 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6481 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6541 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     6601 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6661 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6721 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6781 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     6841 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6901 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6961 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7021 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7081 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7141 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7201 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7261 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7321 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7381 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7441 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7501 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7561 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7621 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7681 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7741 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7801 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7861 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7921 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7981 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8041 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8101 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     8161 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     8221 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     8281 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     8341 aactatgcgg catcagagca gattgtactg agagtgcacc atatggacat attgtcgtta
     8401 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
     8461 tata
//
