LOCUS       sequence-256875-        8257 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of SEC63-2mTagBFP2 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 8257)
  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 8257)
  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: ZJOM135
FEATURES             Location/Qualifiers
     source          1..8257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      4530..4717
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4585..4609)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    4769..4802
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5808..5841
                     /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(5894..5913)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5895..5913)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6000..6017)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6171..6759)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6251..6270)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6930..7721,7722..7790))
                     /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     7553..7572
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7791..7895)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7963..7981
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8019..8041)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8141..8160
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(8241..8257,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(8241..8257,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg tgggattcct gggttaatta
       61 acatcgctag cgttttgacg cgatgtttcc tcttcaaaaa gcttcccaag caacttagat
      121 tttccaatct cctgattatt tcgtttttct ttagaagatt ttttagaatg cctgtgcaaa
      181 gagtgataat ctttaactct cagggaatgg tcgtcacaaa atctgatata taccgaagag
      241 acttcttctt ctacatcctc ttcaagaaca ctaggcctag tctgcatctt attactccta
      301 ccccctaatt tgtcacttct tatagtattt ttatcgtgaa atattatttt aacatcagca
      361 tggtttgctg tgactttctt gatctttcta acggtttctg cgcactgttg gtgttcaact
      421 ttgtggatgc ttccgtcttt tctctttaaa ttattgccaa aaatttcatt gtgatagaac
      481 cttagggcat attttctttt gcttaatttt aaacgagaca aattgttcct ttgcagtaca
      541 tatacccttg gtgattcttg gcaacccttg ggaaaccttg caatcagtag tggtttcttt
      601 ttggaacttc ctggacatag tccagacggt tcatttatca tgtcctcaat gtgtatttcg
      661 gagagggggc tcaagttcca gcgagtatta tattggtgaa tgtgaagttt gtggacatgc
      721 cacctcctct ttaatttccg gccactgaac agttgacaca gtcgaaaata taggtaaaaa
      781 aaatgaccag atttggactt tctcggtgaa ggtggtgctc tcatgcttct tttctatttt
      841 gagttattaa cgtcttagct ttataatgtc acaatctgac ttcattactg acacaatcgc
      901 tcttataacg cgaattggtc actcgttacg gaaaggcgtt ttctgaaaaa tttgtcttga
      961 acacgtcgaa gatcactgaa cgaataaaag atgcgactgg aacaatagtc agttataatt
     1021 gaccaaagaa ggccttcgca acaaaataaa tatactgcat attacagcac aatgcctaca
     1081 aattacgagt atgatgaggc tagtgagacg tggccgtcct tcattttaac ggggctcttg
     1141 atggtcgtcg ggcctatgac actgcttcaa atataccaaa ttttttttgg ggccaatgct
     1201 gaagatggga attcagggaa gagtaaggag tttaatgagg aagttttcaa gaacttgaat
     1261 gaagaataca ccagtgatga aatcaaacaa tttagaagga agtttgataa aaatagtaat
     1321 aagaagtcca aaatatggag caggagaaat attataatta ttgtgggttg gatcttagtt
     1381 gcaattcttc tgcaaaggat taatagtaat gacgcgatta aagacgctgc tacaaaatta
     1441 tttgatcctt atgaaatcct tggtatctct actagtgctt ccgatagaga catcaaatct
     1501 gcttatagaa aattatctgt taaatttcat ccagataaat tagcaaaggg cctaacacct
     1561 gatgagaaaa gtgtgatgga agaaacttat gttcagatta cgaaggctta cgaatccctt
     1621 actgacgaat tggttaggca aaactatttg aaatacggtc atccagatgg cccacaatct
     1681 acttcacatg gtatcgctct accaagattt ttggtagatg gaagtgcatc tccattatta
     1741 gtggtttgtt atgttgcgct actaggttta atcttgccat attttgttag tagatggtgg
     1801 gcaagaacac aatcgtatac taagaaggga atacataatg tgacggcttc taattttgtt
     1861 agtaacttag tcaattacaa gccatctgag attgtcacca cagatttgat cttacactgg
     1921 ttatcatttg ctcatgaatt taaacaattc ttcccggatt tgcaaccaac ggattttgaa
     1981 aaacttttgc aagatcatat taaccgcaga gatagtggta aacttaacaa tgcgaaattt
     2041 agaatagtgg ccaaatgtca ctctttgtta cacggtttat tggatattgc ttgtggattc
     2101 agaaatttag atattgcatt gggtgcaatc aatactttca agtgtattgt tcaggctgta
     2161 ccattaacac caaactgtca aatccttcaa ttgccgaacg tagataaaga gcactttatt
     2221 accaaaaccg gagatattca tacattaggt aaattgttta ctttagaaga tgccaagatt
     2281 ggtgaggttc ttggaataaa ggatcaagca aagttaaacg aaactttgag agttgcatcg
     2341 catattccaa atctaaagat catcaaggca gacttccttg tcccaggtga gaaccaagta
     2401 acaccatcat ctaccccata catttctttg aaagtactgg ttcgttctgc taaacagcca
     2461 ttgataccaa ctagcttaat tcctgaagaa aatttaacag aacctcaaga ttttgaatct
     2521 caaagagatc catttgctat gatgagtaaa cagccactcg tcccatattc ctttgcacca
     2581 tttttcccta caaagagacg tgggagttgg tgctgtctgg taagttctca aaaagatggt
     2641 aaaatacttc aaacgccaat tatcattgaa aagctatctt acaagaactt gaacgatgac
     2701 aaagatttct ttgataagag gataaaaatg gatttaacca aacacgaaaa attcgatata
     2761 aatgattggg aaatcgggac cataaaaatt ccattaggtc agcctgcacc tgaaactgtt
     2821 ggtgatttct tttttagagt aatcgttaaa tccacagatt atttcactac agatttggat
     2881 attaccatga atatgaaagt tcgtgattct cctgcagtgg aacaagtaga ggtgtattct
     2941 gaggaggatg atgagtactc tactgatgac gacgaaaccg aaagtgatga tgaaagtgat
     3001 gctagcgatt atactgatat cgatacggat acagaagctg aagatgatga atcaccagaa
     3061 ctttctcttg aagttgctga agaaatgagc gaactgatca aagagaacat gcacatgaag
     3121 ctgtacatgg aaggcaccgt tgacaaccac cactttaagt gcacctctga gggtgagggt
     3181 aagccctacg aaggcaccca aaccatgcgt atcaaagttg tggagggcgg tccactgcca
     3241 ttcgcttttg acattctggc caccagcttc ctgtacggtt ccaaaacctt cattaaccat
     3301 actcagggca ttcctgattt cttcaaacag agctttccgg aaggtttcac ctgggagcgt
     3361 gtcaccacgt atgaagatgg tggtgtgttg accgccaccc aagatacctc cctgcaagat
     3421 ggctgtctga tctataacgt gaaaattcgt ggcgtcaact ttactagcaa tggtcccgtg
     3481 atgcagaaga aaaccctggg ttgggaggcc tttacagaaa ccctgtatcc tgccgatggt
     3541 ggcctggagg gccgtaacga catggcactg aagctggttg gtggcagcca tttgatcgca
     3601 aatgccaaga ccacttacag aagcaagaaa cccgcaaaaa atctgaagat gccgggtgtt
     3661 tactatgtcg actaccgtct ggaaagaatt aaagaagcaa ataatgagac ttacgtggag
     3721 cagcacgagg ttgcagtcgc gagatattgc gacttgccta gcaagctggg tcataaactg
     3781 aatccattgc cattggatcc aaccggtatg agcgaactga tcaaagagaa catgcacatg
     3841 aagctgtaca tggaaggcac cgttgacaac caccacttta agtgcacctc tgagggtgag
     3901 ggtaagccct acgaaggcac ccaaaccatg cgtatcaaag ttgtggaggg cggtccactg
     3961 ccattcgctt ttgacattct ggccaccagc ttcctgtacg gttccaaaac cttcattaac
     4021 catactcagg gcattcctga tttcttcaaa cagagctttc cggaaggttt cacctgggag
     4081 cgtgtcacca cgtatgaaga tggtggtgtg ttgaccgcca cccaagatac ctccctgcaa
     4141 gatggctgtc tgatctataa cgtgaaaatt cgtggcgtca actttactag caatggtccc
     4201 gtgatgcaga agaaaaccct gggttgggag gcctttacag aaaccctgta tcctgccgat
     4261 ggtggcctgg agggccgtaa cgacatggca ctgaagctgg ttggtggcag ccatttgatc
     4321 gcaaatgcca agaccactta cagaagcaag aaacccgcaa aaaatctgaa gatgccgggt
     4381 gtttactatg tcgactaccg tctggaaaga attaaagaag caaataatga gacttacgtg
     4441 gagcagcacg aggttgcagt cgcgagatat tgcgacttgc ctagcaagct gggtcataaa
     4501 ctgaattgag gcgcgccact tctaaataag cgaatttctt atgatttatg atttttatta
     4561 ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa
     4621 aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat
     4681 agtatgaggt cgctcttatt gaccacacct ctaccggcag atccgctagg gataacaggg
     4741 taatatctgc aggtcgacaa cccttaatat aacttcgtat aatgtatgct atacgaagtt
     4801 attaggtcta gagcggccgc ggaacgatca ttcactatat atatatcaat ttatatatac
     4861 gtatgtgtaa ttgaagaaag atacgttttt tcctctattg agaggcctgc tggatgaata
     4921 gctttacctt ttctaaatcc ttgataccat cagtctctac tcctccactt acatcgacac
     4981 cgatcgcatt tggtaacata ttaatggcaa cagaaacgtt atcaggattc aatccaccag
     5041 cgataatgaa ttttatctcg ggatgacttg cagaccaact ggaaattgca ctccaattca
     5101 atttctcacc agtgccacct tcaccagaat cgaacaacgt cagcacattg tctacgtgtt
     5161 catacaggtc cagtagtaat tcacaatcct gtgggaactg gaacctctta atgattggaa
     5221 ttgaagatgg gatcaaagat ctgtattctt taatatcttc atctccatgt aattgtatca
     5281 catctagatt atattcgtgg tacagttgaa ggacatcatc aacggactga tttctaaaca
     5341 ccccgactag tttagtacct ttcacgttct cttgttggtg aactgcagtt gaaatacctt
     5401 tcgcaacaga tgagtcaatg gttcttttcc taccgggaac acaaatgata cctaagtaat
     5461 cagcaccatc atccacagca gtctttgcag cttcaacggt ttgcaaacca cacactttaa
     5521 cgagcatcac agtagatttc gcaagagggt agctcgctca gagtacccaa gtaaatgatt
     5581 agtaaactga tgtttgatag ttcaattttt caatgaaata accttatatt aaaattgata
     5641 ttactattat acaaaaataa agaataaagg atttgagttt atacataaaa taccattatt
     5701 atttgttcag tgagagatac cggggtatat gggatgtgtg tagtgatacc atgcaatcat
     5761 gtatcaaaca tgggcccggt accgagctct ggtcgagaac ccttaatata acttcgtata
     5821 atgtatgcta tacgaagtta ttaggtgata tcagatccac tagtggccta tgcggccgcg
     5881 gatctgccgg tctccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat
     5941 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6001 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6061 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6121 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6181 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6241 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6301 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6361 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6421 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6481 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6541 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6601 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6661 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6721 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6781 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6841 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6901 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6961 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7021 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7081 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7141 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7201 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7261 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7321 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7381 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7441 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7501 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7561 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7621 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7681 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7741 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7801 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7861 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7921 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     7981 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     8041 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     8101 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     8161 ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta atacataacc
     8221 ttatgtatca tacacatacg atttaggtga cactata
//
