LOCUS       sequence-265905-        8255 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 8255)
  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 8255)
  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..8255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      4531..4718
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4586..4610)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    4770..4803
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5806..5839
                     /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(5892..5911)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5893..5911)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5998..6015)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6169..6757)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6249..6268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6928..7719,7720..7788))
                     /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     7551..7570
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7789..7893)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7961..7979
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8017..8039)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8139..8158
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(8239..8255,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(8239..8255,1)
                     /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 tttttttttg gggccaatgc
     1201 tgaagatggg aattcaggga agagtaagga gtttaatgag gaagttttca agaacttgaa
     1261 tgaagaatac accagtgatg aaatcaaaca atttagaagg aagtttgata aaaatagtaa
     1321 taagaagtcc aaaatatgga gcaggagaaa tattataatt attgtgggtt ggatcttagt
     1381 tgcaattctt ctgcaaagga ttaatagtaa tgacgcgatt aaagacgctg ctacaaaatt
     1441 atttgatcct tatgaaatcc ttggtatctc tactagtgct tccgatagag acatcaaatc
     1501 tgcttataga aaattatctg ttaaatttca tccagataaa ttagcaaagg gcctaacacc
     1561 tgatgagaaa agtgtgatgg aagaaactta tgttcagatt acgaaggctt acgaatccct
     1621 tactgacgaa ttggttaggc aaaactattt gaaatacggt catccagatg gcccacaatc
     1681 tacttcacat ggtatcgctc taccaagatt tttggtagat ggaagtgcat ctccattatt
     1741 agtggtttgt tatgttgcgc tactaggttt aatcttgcca tattttgtta gtagatggtg
     1801 ggcaagaaca caatcgtata ctaagaaggg aatacataat gtgacggctt ctaattttgt
     1861 tagtaactta gtcaattaca agccatctga gattgtcacc acagatttga tcttacactg
     1921 gttatcattt gctcatgaat ttaaacaatt cttcccggat ttgcaaccaa cggattttga
     1981 aaaacttttg caagatcata ttaaccgcag agatagtggt aaacttaaca atgcgaaatt
     2041 tagaatagtg gccaaatgtc actctttgtt acacggttta ttggatattg cttgtggatt
     2101 cagaaattta gatattgcat tgggtgcaat caatactttc aagtgtattg ttcaggctgt
     2161 accattaaca ccaaactgtc aaatccttca attgccgaac gtagataaag agcactttat
     2221 taccaaaacc ggagatattc atacattagg taaattgttt actttagaag atgccaagat
     2281 tggtgaggtt cttggaataa aggatcaagc aaagttaaac gaaactttga gagttgcatc
     2341 gcatattcca aatctaaaga tcatcaaggc agacttcctt gtcccaggtg agaaccaagt
     2401 aacaccatca tctaccccat acatttcttt gaaagtactg gttcgttctg ctaaacagcc
     2461 attgatacca actagcttaa ttcctgaaga aaatttaaca gaacctcaag attttgaatc
     2521 tcaaagagat ccatttgcta tgatgagtaa acagccactc gtcccatatt cctttgcacc
     2581 atttttccct acaaagagac gtgggagttg gtgctgtctg gtaagttctc aaaaagatgg
     2641 taaaatactt caaacgccaa ttatcattga aaagctatct tacaagaact tgaacgatga
     2701 caaagatttc tttgataaga ggataaaaat ggatttaacc aaacacgaaa aattcgatat
     2761 aaatgattgg gaaatcggga ccataaaaat tccattaggt cagcctgcac ctgaaactgt
     2821 tggtgatttc ttttttagag taatcgttaa atccacagat tatttcacta cagatttgga
     2881 tattaccatg aatatgaaag ttcgtgattc tcctgcagtg gaacaagtag aggtgtattc
     2941 tgaggaggat gatgagtact ctactgatga cgacgaaacc gaaagtgatg atgaaagtga
     3001 tgctagcgat tatactgata tcgatacgga tacagaagct gaagatgatg aatcaccaga
     3061 actttctctt gaagttgctg aagaaatgag cgaactgatc aaagagaaca tgcacatgaa
     3121 gctgtacatg gaaggcaccg ttgacaacca ccactttaag tgcacctctg agggtgaggg
     3181 taagccctac gaaggcaccc aaaccatgcg tatcaaagtt gtggagggcg gtccactgcc
     3241 attcgctttt gacattctgg ccaccagctt cctgtacggt tccaaaacct tcattaacca
     3301 tactcagggc attcctgatt tcttcaaaca gagctttccg gaaggtttca cctgggagcg
     3361 tgtcaccacg tatgaagatg gtggtgtgtt gaccgccacc caagatacct ccctgcaaga
     3421 tggctgtctg atctataacg tgaaaattcg tggcgtcaac tttactagca atggtcccgt
     3481 gatgcagaag aaaaccctgg gttgggaggc ctttacagaa accctgtatc ctgccgatgg
     3541 tggcctggag ggccgtaacg acatggcact gaagctggtt ggtggcagcc atttgatcgc
     3601 aaatgccaag accacttaca gaagcaagaa acccgcaaaa aatctgaaga tgccgggtgt
     3661 ttactatgtc gactaccgtc tggaaagaat taaagaagca aataatgaga cttacgtgga
     3721 gcagcacgag gttgcagtcg cgagatattg cgacttgcct agcaagctgg gtcataaact
     3781 gaatccattg ccattggatc caaccggtat gagcgaactg atcaaagaga acatgcacat
     3841 gaagctgtac atggaaggca ccgttgacaa ccaccacttt aagtgcacct ctgagggtga
     3901 gggtaagccc tacgaaggca cccaaaccat gcgtatcaaa gttgtggagg gcggtccact
     3961 gccattcgct tttgacattc tggccaccag cttcctgtac ggttccaaaa ccttcattaa
     4021 ccatactcag ggcattcctg atttcttcaa acagagcttt ccggaaggtt tcacctggga
     4081 gcgtgtcacc acgtatgaag atggtggtgt gttgaccgcc acccaagata cctccctgca
     4141 agatggctgt ctgatctata acgtgaaaat tcgtggcgtc aactttacta gcaatggtcc
     4201 cgtgatgcag aagaaaaccc tgggttggga ggcctttaca gaaaccctgt atcctgccga
     4261 tggtggcctg gagggccgta acgacatggc actgaagctg gttggtggca gccatttgat
     4321 cgcaaatgcc aagaccactt acagaagcaa gaaacccgca aaaaatctga agatgccggg
     4381 tgtttactat gtcgactacc gtctggaaag aattaaagaa gcaaataatg agacttacgt
     4441 ggagcagcac gaggttgcag tcgcgagata ttgcgacttg cctagcaagc tgggtcataa
     4501 actgaattga ggcgcgccac ttctaaataa gcgaatttct tatgatttat gatttttatt
     4561 attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta
     4621 aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta
     4681 tagtatgagg tcgctcttat tgaccacacc tctaccggca gatccgctag ggataacagg
     4741 gtaatatctg caggtcgaca acccttaata taacttcgta taatgtatgc tatacgaagt
     4801 tattaggtct agagcggccg cggaacgatc attcactata tatatatcaa tttatatata
     4861 cgtatgtgta attgaagaaa gatacgtttt ttcctctatt gagaggcctg ctggatgaat
     4921 agctttacct tttctaaatc cttgatacca tcagtctcta ctcctccact tacatcgaca
     4981 ccgatcgcat ttggtaacat attaatggca acagaaacgt tatcaggatt caatccacca
     5041 gcgataatga attttatctc gggatgactt gcagaccaac tggaaattgc actccaattc
     5101 aatttctcac cagtgccacc ttcaccagaa tcgaacaacg tcagcacatt gtctacgtgt
     5161 tcatacaggt ccagtagtaa ttcacaatcc tgtgggaact ggaacctctt aatgattgga
     5221 attgaagatg ggatcaaaga tctgtattct ttaatatctt catctccatg taattgtatc
     5281 acatctagat tatattcgtg gtacagttga aggacatcat caacggactg atttctaaac
     5341 accccgacta gtttagtacc tttcacgttc tcttgttggt gaactgcagt tgaaatacct
     5401 ttcgcaacag atgagtcaat ggttcttttc ctaccgggaa cacaaatgat acctaagtaa
     5461 tcagcaccat catccacagc agtctttgca gcttcaacgg tttgcaaacc acacacttta
     5521 acgagcatca cagtagattt cgcaagaggg tagctcgctc agagtaccca agtaaatgat
     5581 tagtaaactg atgtttgata gttcaatttt tcaatgaaat aaccttatat taaaattgat
     5641 attactatta tacaaaaata aagaataaag gatttgagtt tatacataaa ataccattat
     5701 tatttgttca gtgagagata ctggggtata tgggatgtgt gtagtgatac catgcaatca
     5761 tgtatcaaac atgggcccgg taccgagctc tcgagaaccc ttaatataac ttcgtataat
     5821 gtatgctata cgaagttatt aggtgatatc agatccacta gtggcctatg cggccgcgga
     5881 tctgccggtc tccctatagt gagtcgtatt aatttcgata agccaggtta acctgcatta
     5941 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6001 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6061 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6121 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6181 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6241 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6301 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6361 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6421 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6481 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6541 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6601 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6661 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6721 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6781 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6841 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6901 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6961 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7021 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7081 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7141 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7201 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7261 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7321 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7381 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7441 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7501 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7561 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7621 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7681 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7741 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7801 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7861 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7921 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     7981 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     8041 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     8101 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     8161 gagagtgcac catatggaca tattgtcgtt agaacgcggc tacaattaat acataacctt
     8221 atgtatcata cacatacgat ttaggtgaca ctata
//
