LOCUS       sequence-267526-        6575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of ELO3-mCherry 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 6575)
  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 6575)
  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: ZJOM149
FEATURES             Location/Qualifiers
     source          1..6575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1720..2430
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1865..1883)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2133..2152
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      2460..2647
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2515..2539)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    2699..2732
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3735..3768
                     /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(3821..3840)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3822..3840)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3927..3944)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4098..4686)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4178..4197)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4857..5648,5649..5717))
                     /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     5480..5499
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5718..5822)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5890..5908
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5946..5968)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6068..6087
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6168..6186
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6168..6185
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 cctgtactgc tacgaggatt atcgcgggta tgatacagtt gcgtcgttag atttctgggg
       61 ttaagatagc aacgtatata gactttcctt attttcttta gaataaaaag cataactaaa
      121 gttgtaaaag gagcagaatt atgaataata aacattgatg acaatagtgg aaaacgacat
      181 aaataataat tttataatat aataatgata attcaccgtc agagggattt gtatgcaaat
      241 gattgatgac gcgagaaaaa aaaacgcgaa atttttcttc ccaaagcttc gaagttttta
      301 ttccgtccac aaagtgaaaa attttcagaa aattttccag actgtgaata aacaaaaggt
      361 tggcttacgc atttggcttg tctttttttt ttttcacagg ttcttagtaa ataattgtat
      421 aaaatattta tctggagtct tctgtttgtt gtttagcctt gtttgaagtt ttatttaact
      481 tattacccca gttaggtact gtgtattttc cgcatatctg gtgtcttttt atttggtcat
      541 tgattggttt ttgacagctc ttcactcgct tcaagcttac ttctagttta tttattcggc
      601 ttttttccgt ttgtttacga aacataaaca gtcatgaaca ctaccacatc tactgttata
      661 gcagcagttg ccgaccagtt ccagtctttg aactcttctt cttcatgttt cttgaaggtt
      721 catgttcctt ccattgagaa cccattcggt attgaattat ggccaatttt ctccaaagtg
      781 tttgaatact ttagtggcta tccagctgag caattcgagt ttattcacaa taagactttc
      841 ttggctaacg ggtatcatgc tgttagtatt attatcgttt attacattat tatctttggt
      901 ggccaagcta tcttacgcgc cttgaacgcc tctccattaa agtttaaatt gcttttcgag
      961 atacacaact tgtttttgac ttctatttct ctagttttat ggttgctgat gttagaacag
     1021 ttggttccta tggtttatca caacggtcta ttctggtcta tctgctctaa ggaagccttc
     1081 gcaccaaaat tagttactct ttactatttg aactatttga ccaaattcgt agaattgatt
     1141 gacactgtgt ttttagtttt gagaagaaag aagttattgt ttttgcacac ttaccatcac
     1201 ggtgccaccg ctttgttgtg ctacactcaa ttaattggtc gtacttctgt tgaatgggta
     1261 gttatcctac taaacttggg tgttcacgtt atcatgtact ggtactactt cttgagttca
     1321 tgtggtatta gagtttggtg gaagcaatgg gtcactagat tccaaattat tcaatttttg
     1381 attgacttgg tatttgttta ctttgctacc tatacattct atgctcacaa atacttggac
     1441 ggtattttac caaacaaggg tacttgttat ggtactcagg ctgctgctgc ttatgggtat
     1501 ttgattctaa catcttattt gcttttgttt atttccttct acatccaatc ttacaagaaa
     1561 ggtggtaaaa agacagtcaa gaaggaatct gaagtttccg gctccgttgc atccggttct
     1621 tctactggtg tcaagacctc taacaccaag gtctcttcca ggaaagctgc ttcgtacgct
     1681 gcaggtcgtg ggattcctgg gttaattaac atcgctagca tggtgagcaa gggcgaggag
     1741 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     1801 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     1861 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     1921 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     1981 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     2041 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     2101 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     2161 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     2221 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     2281 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     2341 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     2401 accggcggca tggacgagct gtacaagtag cctaggtgag gcgcgccact tctaaataag
     2461 cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat
     2521 acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt
     2581 tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct
     2641 ctaccggcag atccgctagg gataacaggg taatatctgc aggtcgacaa cccttaatat
     2701 aacttcgtat aatgtatgct atacgaagtt attaggtcta gagcggccgc ggaacgatca
     2761 ttcactatat atatatcaat ttatatatac gtatgtgtaa ttgaagaaag atacgttttt
     2821 tcctctattg agaggcctgc tggatgaata gctttacctt ttctaaatcc ttgataccat
     2881 cagtctctac tcctccactt acatcgacac cgatcgcatt tggtaacata ttaatggcaa
     2941 cagaaacgtt atcaggattc aatccaccag cgataatgaa ttttatctcg ggatgacttg
     3001 cagaccaact ggaaattgca ctccaattca atttctcacc agtgccacct tcaccagaat
     3061 cgaacaacgt cagcacattg tctacgtgtt catacaggtc cagtagtaat tcacaatcct
     3121 gtgggaactg gaacctctta atgattggaa ttgaagatgg gatcaaagat ctgtattctt
     3181 taatatcttc atctccatgt aattgtatca catctagatt atattcgtgg tacagttgaa
     3241 ggacatcatc aacggactga tttctaaaca ccccgactag tttagtacct ttcacgttct
     3301 cttgttggtg aactgcagtt gaaatacctt tcgcaacaga tgagtcaatg gttcttttcc
     3361 taccgggaac acaaatgata cctaagtaat cagcaccatc atccacagca gtctttgcag
     3421 cttcaacggt ttgcaaacca cacactttaa cgagcatcac agtagatttc gcaagagggt
     3481 agctcgctca gagtacccaa gtaaatgatt agtaaactga tgtttgatag ttcaattttt
     3541 caatgaaata accttatatt aaaattgata ttactattat acaaaaataa agaataaagg
     3601 atttgagttt atacataaaa taccattatt atttgttcag tgagagatac tggggtatat
     3661 gggatgtgtg tagtgatacc atgcaatcat gtatcaaaca tgggcccggt accgagctct
     3721 cgagaaccct taatataact tcgtataatg tatgctatac gaagttatta ggtgatatca
     3781 gatccactag tggcctatgc ggccgcggat ctgccggtct ccctatagtg agtcgtatta
     3841 atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3901 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3961 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4021 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4081 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4141 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4201 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4261 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4321 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4381 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4441 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4501 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4561 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4621 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4681 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4741 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4801 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4861 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4921 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4981 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5041 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5101 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5161 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5221 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5281 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5341 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5401 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5461 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5521 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5581 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5641 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5701 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5761 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5821 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     5881 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     5941 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     6001 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     6061 aactatgcgg catcagagca gattgtactg agagtgcacc atatggacat attgtcgtta
     6121 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
     6181 tatagaacgc ggccgccagc tgaagcttgt aaaaagggtt ttcagatcgt tagcatttca
     6241 cagaatgctc atgaatcctt gttagatccg gccgatacat cattagagtt tgcactaagg
     6301 gatactttga aaccaatggc aatctatcgt gtgggaaata tgtttttgct gtgttacaca
     6361 gaatttgcct tctttgtcaa taatcagggt tggagaaaga aagaatcgca tattattcat
     6421 tgggagggag agccccaaaa atttgcaatt tggtacccgt acatcttggc attcgacagc
     6481 aacttcattg agatcagaaa gattgaaact ggcgaattaa ttcgttgcgt ccttgctgat
     6541 aagattcgac ttctacaaac aagtactcaa gaaat
//
