LOCUS       sequence-265883-        8461 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 8461)
  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 8461)
  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..8461
                     /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      4754..4941
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4809..4833)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    4993..5026
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    6029..6062
                     /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(6115..6134)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6116..6134)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6221..6238)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6392..6980)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6472..6491)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7151..7942,7943..8011))
                     /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     7774..7793
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8012..8116)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8184..8202
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8240..8262)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8362..8381
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaggtga cactatagaa cgcggccgcc agctgaagct tcgtacgctg caggtcgtgg
       61 gattcctggg ttaattaaca tcgctagctt caccgttaac atctgtcgta tcgaacaaat
      121 cggaaaagtc cgtgcctttg gtttctagtt acgtagaaga actgcgacta aagtattaca
      181 agacttctaa ctttttacaa gcgcctccta acttgccggt tgctctaaaa caaaaaaata
      241 atttgattca acccaagaat attaaggtca agttgaggac aagttctaag caaattggta
      301 tcaagcatgg taaagttaaa cagaaattac tggcgttgga gacaagaaat gaagaatcag
      361 atggaaccgc tacagggctg aaaaataaaa tcaatgttga ccacacaaag gaattccata
      421 aattgttagg gaaggaaaat gaaactggct ctatttcaaa aaaagaaggc actgacgcag
      481 aacaagccga agactacctc aaggacatac ccggtgatga agcatataac agtgatgata
      541 taatggcgcc tttacgagaa aaaagagggc aaaacggttc tgttgattcc gtgtcaagat
      601 ctaacacggt agtcagctac tacacaaggt ctcagaacag aatgagaagt ggaacactgg
      661 ataatgatta cgtgaacaga caaaagcttc ctacacatat ctctcttcaa gattatcgtg
      721 atgctaatgc tagaagtaat atatcgcgtc aggactctgt ctccacaacg aactctgatg
      781 tggtagacct cagctattct ctggggcatg gcttgcgtgt ggcaaaccct gattcagacc
      841 cagaatgata tccttctctg tagttttgta gatgtcatat atgtacgttt atggaacagc
      901 attttagaaa agtattaccc agcttatcac ttcgtttttt ttttctttcc gtgtactcgc
      961 tacacgtaga aagaaatcag aaaacaacag ctcgacaagt gaaatttgac gttcattaag
     1021 actcagttaa gattgccttg agaataaaca aaagtaatca cagttaacta ttgaacaaga
     1081 gtgcacctat gacttcactt tcttcacagc cagcgtacac gttggttttc gatccttctc
     1141 cgagtatgga gacttactcg agtaccgatt ttcagaaagc tcttgaaaag ggatctgatg
     1201 aacaaaaaat tgacacgatg aaatcaattt tagttacaat gctggaagga aatccaatgc
     1261 ctgaattgtt gatgcacata ataagatttg tcatgccttc taaaaataag gaattaaaaa
     1321 agcttttgta cttctactgg gaaattgttc ccaaactagc tgaagatgga aaattgagac
     1381 atgaaatgat tcttgtctgt aatgccattc aacacgattt gcaacatcct aatgaatata
     1441 ttagaggtaa cacattaagg tttttaacga aattgagaga ggccgaactc ttagaacaga
     1501 tggttccctc tgtcttagcg tgcttggaat accgtcatgc atatgttcgt aagtatgcaa
     1561 tcctagcagt tttctccatt ttcaaggtca gcgaacattt acttcccgat gctaaagaaa
     1621 tcatcaattc gttcatagta gctgaaactg atccaatatg taaaagaaat gcatttattg
     1681 ggttagctga attagatcgt gaaaatgcct tacactattt agagaacaat attgctgata
     1741 tagaaaacct agacccttta ttacaagctg tctttgttca atttatcaga caagatgcaa
     1801 acaggacccc tgctttgaaa gcccaatata tcgaattatt gatggaactg ctttcgacca
     1861 cgacttccga tgaagtcatc ttcgagaccg cattagccct aactgtgttg tctgccaatc
     1921 caaatgtctt ggttcctgcg gttaacaaat tgattgactt ggccgtcaag gtttctgata
     1981 ataacattaa gttaattgtt ctagaccgta ttcaagacat caatgctaat aacgtaggtg
     2041 ctttggaaga gttaaccctg gatattttga gagtcttgaa tgcagaagat ttagacgttc
     2101 gttcaaaggc gcttgatatt tcaatggact tggccacatc cagaaatgct gaagatgttg
     2161 ttcagctttt gaagaaagag ctgcaaacaa ccgtaaataa cccagatcaa gacaaggcaa
     2221 tgcagtacag acaattgtta ataaaaacta ttcgtaccgt ggctgtaaac tttgtagaaa
     2281 tggcagcaag tgttgtttcg ctattattag atttcatcgg tgatttaaac tcggttgccg
     2341 ccagtggtat cattgccttt atcaaagaag tgatcgaaaa atacccacaa cttagagcca
     2401 atatccttga aaacatggtt caaacgctag acaaagtgag atctgctaaa gcttaccgcg
     2461 gtgcattatg gattatgggt gagtatgctg aaggagaaag tgagatacaa cattgttgga
     2521 agcacattcg taacagcgta ggtgaagttc ctatccttca atcagaaatc aaaaagttaa
     2581 cacaaaacca agaacacacc gaagaaaatg aggttgacgc taccgccaag ccaactggtc
     2641 cagttattct accagacggt acgtatgcca ctgaaagcgc tttcgatgtg aagacttctc
     2701 aaaagtcagt taccgatgaa gaacgtgatt ctagacctcc aattcgccgg tttgttttaa
     2761 gtggtgattt ctacacagct gccattctgg ccaacaccat cattaaactt gttttaaaat
     2821 tcgaaaacgt ttccaagaac aaaactgtca tcaatgctct aaaggcggaa gctttactaa
     2881 ttttagttag tattgtaaga gtgggtcaaa gctctttggt ggagaaaaaa attgatgaag
     2941 attctttaga gagagttatg acatctattt ctattttatt ggatgaagtt aatcctgagg
     3001 aaaagaagga agaagttaaa cttctggagg ttgcattcct ggacaccacc aaatcctcat
     3061 tcaagagaca aattgaaatt gcaaagaaga acaagcataa gagagcatta aaagacagtt
     3121 gcaaaaacat cgaaccaatt gatacgccga tttctttcag gcaatttgct ggtgtggatt
     3181 ctactaatgt gcaaaaagat agtattgaag aagatttaca actggcaatg aaaggagatg
     3241 caatccacgc tactagcagc tctagtattt cgaagctgaa gaagatagta cctttatgtg
     3301 gcttttctga tccagtttac gccgaggctt gtattacaaa caatcaattt gacgtcgtat
     3361 tagatgttct tcttgttaat caaacgaaag aaacattgaa aaacctacat gtgcaatttg
     3421 caactcttgg tgatttgaag attattgaca caccacagaa gaccaacgtg attcctcatg
     3481 gcttccacaa attcactgtt actgtcaaag tttcctctgc tgacacaggt gtcattttcg
     3541 gtaatattat ttatgatggt gcgcatggtg aagatgctcg ttatgttatt ttaaacgacg
     3601 ttcatgttga cattatggat tatatcaaac cagccactgc tgacgatgaa catttccgta
     3661 ccatgtggaa tgcatttgag tgggagaaca aaatatcggt caaatcacaa ctaccaacat
     3721 tgcatgctta tttgagagaa ctggtcaagg gaactaatat gggtattcta acaccatcag
     3781 agtcgttggg agaagatgat tgtaggttct taagttgtaa tctgtatgcg aagtcgtcct
     3841 ttggtgaaga tgccctagcc aacttgtgta tcgaaaagga ttccaaaacc aatgatgtca
     3901 taggttatgt tcgtatccga tcaaagggac aaggtttggc tctgtcccta ggtgacagag
     3961 tggcattgat tgctaagaag accaataaac ttgctctcac tcatgttctt tctcttgaag
     4021 ttgctgaaga aatgagcgaa ctgatcaaag agaacatgca catgaagctg tacatggaag
     4081 gcaccgttga caaccaccac tttaagtgca cctctgaggg tgagggtaag ccctacgaag
     4141 gcacccaaac catgcgtatc aaagttgtgg agggcggtcc actgccattc gcttttgaca
     4201 ttctggccac cagcttcctg tacggttcca aaaccttcat taaccatact cagggcattc
     4261 ctgatttctt caaacagagc tttccggaag gtttcacctg ggagcgtgtc accacgtatg
     4321 aagatggtgg tgtgttgacc gccacccaag atacctccct gcaagatggc tgtctgatct
     4381 ataacgtgaa aattcgtggc gtcaacttta ctagcaatgg tcccgtgatg cagaagaaaa
     4441 ccctgggttg ggaggccttt acagaaaccc tgtatcctgc cgatggtggc ctggagggcc
     4501 gtaacgacat ggcactgaag ctggttggtg gcagccattt gatcgcaaat gccaagacca
     4561 cttacagaag caagaaaccc gcaaaaaatc tgaagatgcc gggtgtttac tatgtcgact
     4621 accgtctgga aagaattaaa gaagcaaata atgagactta cgtggagcag cacgaggttg
     4681 cagtcgcgag atattgcgac ttgcctagca agctgggtca taaactgaat tgaggcgcgc
     4741 cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat aagttataaa
     4801 aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt
     4861 cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagtatg aggtcgctct
     4921 tattgaccac acctctaccg gcagatccgc tagggataac agggtaatat ctgcaggtcg
     4981 acaaccctta atataacttc gtataatgta tgctatacga agttattagg tctagagcgg
     5041 ccgcggaacg atcattcact atatatatat caatttatat atacgtatgt gtaattgaag
     5101 aaagatacgt tttttcctct attgagaggc ctgctggatg aatagcttta ccttttctaa
     5161 atccttgata ccatcagtct ctactcctcc acttacatcg acaccgatcg catttggtaa
     5221 catattaatg gcaacagaaa cgttatcagg attcaatcca ccagcgataa tgaattttat
     5281 ctcgggatga cttgcagacc aactggaaat tgcactccaa ttcaatttct caccagtgcc
     5341 accttcacca gaatcgaaca acgtcagcac attgtctacg tgttcataca ggtccagtag
     5401 taattcacaa tcctgtggga actggaacct cttaatgatt ggaattgaag atgggatcaa
     5461 agatctgtat tctttaatat cttcatctcc atgtaattgt atcacatcta gattatattc
     5521 gtggtacagt tgaaggacat catcaacgga ctgatttcta aacaccccga ctagtttagt
     5581 acctttcacg ttctcttgtt ggtgaactgc agttgaaata cctttcgcaa cagatgagtc
     5641 aatggttctt ttcctaccgg gaacacaaat gatacctaag taatcagcac catcatccac
     5701 agcagtcttt gcagcttcaa cggtttgcaa accacacact ttaacgagca tcacagtaga
     5761 tttcgcaaga gggtagctcg ctcagagtac ccaagtaaat gattagtaaa ctgatgtttg
     5821 atagttcaat ttttcaatga aataacctta tattaaaatt gatattacta ttatacaaaa
     5881 ataaagaata aaggatttga gtttatacat aaaataccat tattatttgt tcagtgagag
     5941 atactggggt atatgggatg tgtgtagtga taccatgcaa tcatgtatca aacatgggcc
     6001 cggtaccgag ctctcgagaa cccttaatat aacttcgtat aatgtatgct atacgaagtt
     6061 attaggtgat atcagatcca ctagtggcct atgcggccgc ggatctgccg gtctccctat
     6121 agtgagtcgt attaatttcg ataagccagg ttaacctgca ttaatgaatc ggccaacgcg
     6181 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6241 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6301 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6361 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6421 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6481 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6541 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6601 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6661 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6721 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6781 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6841 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6901 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6961 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7021 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7081 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7141 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7201 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7261 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7321 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7381 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7441 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7501 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7561 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7621 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7681 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7741 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7801 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7861 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7921 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7981 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8041 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8101 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     8161 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     8221 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     8281 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     8341 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgg
     8401 acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac
     8461 g
//
