LOCUS       sequence-256876-        6580 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 6580)
  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 6580)
  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..6580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2853..3040
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2908..2932)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    3092..3125
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4131..4164
                     /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(4217..4236)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4218..4236)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4323..4340)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4494..5082)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4574..4593)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5253..6044,6045..6113))
                     /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     5876..5895
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6114..6218)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6286..6304
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6342..6364)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6464..6483
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(6564..6580,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(6564..6580,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg tgggattcct gggttaatta
       61 acatcgctag cgtaaaaagg gttttcagat cgttagcatt tcacagaatg ctcatgaatc
      121 cttgttagat ccggccgata catcattaga gtttgcacta agggatactt tgaaaccaat
      181 ggcaatctat cgtgtgggaa atatgttttt gctgtgttac acagaatttg ccttctttgt
      241 caataatcag ggttggagaa agaaagaatc gcatattatt cattgggagg gagagcccca
      301 aaaatttgca atttggtacc cgtacatctt ggcattcgac agcaacttca ttgagatcag
      361 aaagattgaa actggcgaat taattcgttg cgtccttgct gataagattc gacttctaca
      421 aacaagtact caagaaatcc tgtactgcta cgaggattat cgcgggtatg atacagttgc
      481 gtcgttagat ttctggggtt aagatagcaa cgtatataga ctttccttat tttctttaga
      541 ataaaaagca taactaaagt tgtaaaagga gcagaattat gaataataaa cattgatgac
      601 aatagtggaa aacgacataa ataataattt tataatataa taatgataat tcaccgtcag
      661 agggatttgt atgcaaatga ttgatgacgc gagaaaaaaa aacgcgaaat ttttcttccc
      721 aaagcttcga agtttttatt ccgtccacaa agtgaaaaat tttcagaaaa ttttccagac
      781 tgtgaataaa caaaaggttg gcttacgcat ttggcttgtc tttttttttt ttcacaggtt
      841 cttagtaaat aattgtataa aatatttatc tggagtcttc tgtttgttgt ttagccttgt
      901 ttgaagtttt atttaactta ttaccccagt taggtactgt gtattttccg catatctggt
      961 gtctttttat ttggtcattg attggttttt gacagctctt cactcgcttc aagcttactt
     1021 ctagtttatt tattcggctt ttttccgttt gtttacgaaa cataaacagt catgaacact
     1081 accacatcta ctgttatagc agcagttgcc gaccagttcc agtctttgaa ctcttcttct
     1141 tcatgtttct tgaaggttca tgttccttcc attgagaacc cattcggtat tgaattatgg
     1201 ccaattttct ccaaagtgtt tgaatacttt agtggctatc cagctgagca attcgagttt
     1261 attcacaata agactttctt ggctaacggg tatcatgctg ttagtattat tatcgtttat
     1321 tacattatta tctttggtgg ccaagctatc ttacgcgcct tgaacgcctc tccattaaag
     1381 tttaaattgc ttttcgagat acacaacttg tttttgactt ctatttctct agttttatgg
     1441 ttgctgatgt tagaacagtt ggttcctatg gtttatcaca acggtctatt ctggtctatc
     1501 tgctctaagg aagccttcgc accaaaatta gttactcttt actatttgaa ctatttgacc
     1561 aaattcgtag aattgattga cactgtgttt ttagttttga gaagaaagaa gttattgttt
     1621 ttgcacactt accatcacgg tgccaccgct ttgttgtgct acactcaatt aattggtcgt
     1681 acttctgttg aatgggtagt tatcctacta aacttgggtg ttcacgttat catgtactgg
     1741 tactacttct tgagttcatg tggtattaga gtttggtgga agcaatgggt cactagattc
     1801 caaattattc aatttttgat tgacttggta tttgtttact ttgctaccta tacattctat
     1861 gctcacaaat acttggacgg tattttacca aacaagggta cttgttatgg tactcaggct
     1921 gctgctgctt atgggtattt gattctaaca tcttatttgc ttttgtttat ttccttctac
     1981 atccaatctt acaagaaagg tggtaaaaag acagtcaaga aggaatctga agtttccggc
     2041 tccgttgcat ccggttcttc tactggtgtc aagacctcta acaccaaggt ctcttccagg
     2101 aaagctcttt ctcttgaagt tgctgaagaa atgagcgaac tgatcaaaga gaacatgcac
     2161 atgaagctgt acatggaagg caccgttgac aaccaccact ttaagtgcac ctctgagggt
     2221 gagggtaagc cctacgaagg cacccaaacc atgcgtatca aagttgtgga gggcggtcca
     2281 ctgccattcg cttttgacat tctggccacc agcttcctgt acggttccaa aaccttcatt
     2341 aaccatactc agggcattcc tgatttcttc aaacagagct ttccggaagg tttcacctgg
     2401 gagcgtgtca ccacgtatga agatggtggt gtgttgaccg ccacccaaga tacctccctg
     2461 caagatggct gtctgatcta taacgtgaaa attcgtggcg tcaactttac tagcaatggt
     2521 cccgtgatgc agaagaaaac cctgggttgg gaggccttta cagaaaccct gtatcctgcc
     2581 gatggtggcc tggagggccg taacgacatg gcactgaagc tggttggtgg cagccatttg
     2641 atcgcaaatg ccaagaccac ttacagaagc aagaaacccg caaaaaatct gaagatgccg
     2701 ggtgtttact atgtcgacta ccgtctggaa agaattaaag aagcaaataa tgagacttac
     2761 gtggagcagc acgaggttgc agtcgcgaga tattgcgact tgcctagcaa gctgggtcat
     2821 aaactgaatt gaggcgcgcc acttctaaat aagcgaattt cttatgattt atgattttta
     2881 ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt
     2941 taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg
     3001 tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct agggataaca
     3061 gggtaatatc tgcaggtcga caacccttaa tataacttcg tataatgtat gctatacgaa
     3121 gttattaggt ctagagcggc cgcggaacga tcattcacta tatatatatc aatttatata
     3181 tacgtatgtg taattgaaga aagatacgtt ttttcctcta ttgagaggcc tgctggatga
     3241 atagctttac cttttctaaa tccttgatac catcagtctc tactcctcca cttacatcga
     3301 caccgatcgc atttggtaac atattaatgg caacagaaac gttatcagga ttcaatccac
     3361 cagcgataat gaattttatc tcgggatgac ttgcagacca actggaaatt gcactccaat
     3421 tcaatttctc accagtgcca ccttcaccag aatcgaacaa cgtcagcaca ttgtctacgt
     3481 gttcatacag gtccagtagt aattcacaat cctgtgggaa ctggaacctc ttaatgattg
     3541 gaattgaaga tgggatcaaa gatctgtatt ctttaatatc ttcatctcca tgtaattgta
     3601 tcacatctag attatattcg tggtacagtt gaaggacatc atcaacggac tgatttctaa
     3661 acaccccgac tagtttagta cctttcacgt tctcttgttg gtgaactgca gttgaaatac
     3721 ctttcgcaac agatgagtca atggttcttt tcctaccggg aacacaaatg atacctaagt
     3781 aatcagcacc atcatccaca gcagtctttg cagcttcaac ggtttgcaaa ccacacactt
     3841 taacgagcat cacagtagat ttcgcaagag ggtagctcgc tcagagtacc caagtaaatg
     3901 attagtaaac tgatgtttga tagttcaatt tttcaatgaa ataaccttat attaaaattg
     3961 atattactat tatacaaaaa taaagaataa aggatttgag tttatacata aaataccatt
     4021 attatttgtt cagtgagaga taccggggta tatgggatgt gtgtagtgat accatgcaat
     4081 catgtatcaa acatgggccc ggtaccgagc tctggtcgag aacccttaat ataacttcgt
     4141 ataatgtatg ctatacgaag ttattaggtg atatcagatc cactagtggc ctatgcggcc
     4201 gcggatctgc cggtctccct atagtgagtc gtattaattt cgataagcca ggttaacctg
     4261 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4321 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4381 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4441 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4501 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4561 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4621 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4681 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4741 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4801 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4861 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4921 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4981 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5041 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5101 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5161 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5221 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5281 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5341 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5401 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5461 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5521 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5581 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5641 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5701 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5761 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5821 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5881 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5941 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6001 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6061 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6121 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6181 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6241 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     6301 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     6361 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     6421 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     6481 gtactgagag tgcaccatat ggacatattg tcgttagaac gcggctacaa ttaatacata
     6541 accttatgta tcatacacat acgatttagg tgacactata
//
