LOCUS       sequence-265888-        6577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of ELO3-mTagBFP2 as an additional copy in genome, use 
            auxotrophic marker TRP1(Kluyveromyces lactis). Marker for yeast 
            endoplasmic reticulum (ER)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6577)
  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 6577)
  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: ZJOM136
FEATURES             Location/Qualifiers
     source          1..6577
                     /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      2870..3057
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2925..2949)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    3109..3142
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4145..4178
                     /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(4231..4250)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4232..4250)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4337..4354)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4508..5096)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4588..4607)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5267..6058,6059..6127))
                     /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     5890..5909
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6128..6232)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6300..6318
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6356..6378)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6478..6497
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaggtga cactatagaa cgcggccgcc agctgaagct tcgtacgctg caggtcgtgg
       61 gattcctggg ttaattaaca tcgctagcgt aaaaagggtt ttcagatcgt tagcatttca
      121 cagaatgctc atgaatcctt gttagatccg gccgatacat cattagagtt tgcactaagg
      181 gatactttga aaccaatggc aatctatcgt gtgggaaata tgtttttgct gtgttacaca
      241 gaatttgcct tctttgtcaa taatcagggt tggagaaaga aagaatcgca tattattcat
      301 tgggagggag agccccaaaa atttgcaatt tggtacccgt acatcttggc attcgacagc
      361 aacttcattg agatcagaaa gattgaaact ggcgaattaa ttcgttgcgt ccttgctgat
      421 aagattcgac ttctacaaac aagtactcaa gaaatcctgt actgctacga ggattatcgc
      481 gggtatgata cagttgcgtc gttagatttc tggggttaag atagcaacgt atatagactt
      541 tccttatttt ctttagaata aaaagcataa ctaaagttgt aaaaggagca gaattatgaa
      601 taataaacat tgatgacaat agtggaaaac gacataaata ataattttat aatataataa
      661 tgataattca ccgtcagagg gatttgtatg caaatgattg atgacgcgag aaaaaaaaac
      721 gcgaaatttt tcttcccaaa gcttcgaagt ttttattccg tccacaaagt gaaaaatttt
      781 cagaaaattt tccagactgt gaataaacaa aaggttggct tacgcatttg gcttgtcttt
      841 tttttttttc acaggttctt agtaaataat tgtataaaat atttatctgg agtcttctgt
      901 ttgttgttta gccttgtttg aagttttatt taacttatta ccccagttag gtactgtgta
      961 ttttccgcat atctggtgtc tttttatttg gtcattgatt ggtttttgac agctcttcac
     1021 tcgcttcaag cttacttcta gtttatttat tcggcttttt tccgtttgtt tacgaaacat
     1081 aaacagtcat gaacactacc acatctactg ttatagcagc agttgccgac cagttccagt
     1141 ctttgaactc ttcttcttca tgtttcttga aggttcatgt tccttccatt gagaacccat
     1201 tcggtattga attatggcca attttctcca aagtgtttga atactttagt ggctatccag
     1261 ctgagcaatt cgagtttatt cacaataaga ctttcttggc taacgggtat catgctgtta
     1321 gtattattat cgtttattac attattatct ttggtggcca agctatctta cgcgccttga
     1381 acgcctctcc attaaagttt aaattgcttt tcgagataca caacttgttt ttgacttcta
     1441 tttctctagt tttatggttg ctgatgttag aacagttggt tcctatggtt tatcacaacg
     1501 gtctattctg gtctatctgc tctaaggaag ccttcgcacc aaaattagtt actctttact
     1561 atttgaacta tttgaccaaa ttcgtagaat tgattgacac tgtgttttta gttttgagaa
     1621 gaaagaagtt attgtttttg cacacttacc atcacggtgc caccgctttg ttgtgctaca
     1681 ctcaattaat tggtcgtact tctgttgaat gggtagttat cctactaaac ttgggtgttc
     1741 acgttatcat gtactggtac tacttcttga gttcatgtgg tattagagtt tggtggaagc
     1801 aatgggtcac tagattccaa attattcaat ttttgattga cttggtattt gtttactttg
     1861 ctacctatac attctatgct cacaaatact tggacggtat tttaccaaac aagggtactt
     1921 gttatggtac tcaggctgct gctgcttatg ggtatttgat tctaacatct tatttgcttt
     1981 tgtttatttc cttctacatc caatcttaca agaaaggtgg taaaaagaca gtcaagaagg
     2041 aatctgaagt ttccggctcc gttgcatccg gttcttctac tggtgtcaag acctctaaca
     2101 ccaaggtctc ttccaggaaa gctctttctc ttgaagttgc tgaagaaatg agcgaactga
     2161 tcaaagagaa catgcacatg aagctgtaca tggaaggcac cgttgacaac caccacttta
     2221 agtgcacctc tgagggtgag ggtaagccct acgaaggcac ccaaaccatg cgtatcaaag
     2281 ttgtggaggg cggtccactg ccattcgctt ttgacattct ggccaccagc ttcctgtacg
     2341 gttccaaaac cttcattaac catactcagg gcattcctga tttcttcaaa cagagctttc
     2401 cggaaggttt cacctgggag cgtgtcacca cgtatgaaga tggtggtgtg ttgaccgcca
     2461 cccaagatac ctccctgcaa gatggctgtc tgatctataa cgtgaaaatt cgtggcgtca
     2521 actttactag caatggtccc gtgatgcaga agaaaaccct gggttgggag gcctttacag
     2581 aaaccctgta tcctgccgat ggtggcctgg agggccgtaa cgacatggca ctgaagctgg
     2641 ttggtggcag ccatttgatc gcaaatgcca agaccactta cagaagcaag aaacccgcaa
     2701 aaaatctgaa gatgccgggt gtttactatg tcgactaccg tctggaaaga attaaagaag
     2761 caaataatga gacttacgtg gagcagcacg aggttgcagt cgcgagatat tgcgacttgc
     2821 ctagcaagct gggtcataaa ctgaattgag gcgcgccact tctaaataag cgaatttctt
     2881 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta
     2941 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt
     3001 caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct ctaccggcag
     3061 atccgctagg gataacaggg taatatctgc aggtcgacaa cccttaatat aacttcgtat
     3121 aatgtatgct atacgaagtt attaggtcta gagcggccgc ggaacgatca ttcactatat
     3181 atatatcaat ttatatatac gtatgtgtaa ttgaagaaag atacgttttt tcctctattg
     3241 agaggcctgc tggatgaata gctttacctt ttctaaatcc ttgataccat cagtctctac
     3301 tcctccactt acatcgacac cgatcgcatt tggtaacata ttaatggcaa cagaaacgtt
     3361 atcaggattc aatccaccag cgataatgaa ttttatctcg ggatgacttg cagaccaact
     3421 ggaaattgca ctccaattca atttctcacc agtgccacct tcaccagaat cgaacaacgt
     3481 cagcacattg tctacgtgtt catacaggtc cagtagtaat tcacaatcct gtgggaactg
     3541 gaacctctta atgattggaa ttgaagatgg gatcaaagat ctgtattctt taatatcttc
     3601 atctccatgt aattgtatca catctagatt atattcgtgg tacagttgaa ggacatcatc
     3661 aacggactga tttctaaaca ccccgactag tttagtacct ttcacgttct cttgttggtg
     3721 aactgcagtt gaaatacctt tcgcaacaga tgagtcaatg gttcttttcc taccgggaac
     3781 acaaatgata cctaagtaat cagcaccatc atccacagca gtctttgcag cttcaacggt
     3841 ttgcaaacca cacactttaa cgagcatcac agtagatttc gcaagagggt agctcgctca
     3901 gagtacccaa gtaaatgatt agtaaactga tgtttgatag ttcaattttt caatgaaata
     3961 accttatatt aaaattgata ttactattat acaaaaataa agaataaagg atttgagttt
     4021 atacataaaa taccattatt atttgttcag tgagagatac tggggtatat gggatgtgtg
     4081 tagtgatacc atgcaatcat gtatcaaaca tgggcccggt accgagctct cgagaaccct
     4141 taatataact tcgtataatg tatgctatac gaagttatta ggtgatatca gatccactag
     4201 tggcctatgc ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa
     4261 gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4321 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4381 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4441 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4501 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4561 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4621 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4681 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4741 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4801 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4861 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4921 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4981 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5041 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5101 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5161 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5221 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5281 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5341 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5401 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5461 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5521 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5581 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5641 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5701 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5761 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5821 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5881 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5941 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6001 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6061 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6121 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6181 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6241 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     6301 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     6361 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     6421 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     6481 catcagagca gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct
     6541 acaattaata cataacctta tgtatcatac acatacg
//
