LOCUS       sequence-256877-        7120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of NAB2-mTagBFP2 as an additional copy in genome, use 
            auxotrophic marker TRP1(Kluyveromyces lactis). Marker for yeast 
            nucleus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7120)
  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 7120)
  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: ZJOM119
FEATURES             Location/Qualifiers
     source          1..7120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      3393..3580
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3448..3472)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    3632..3665
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4671..4704
                     /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(4757..4776)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4758..4776)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4863..4880)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5034..5622)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5114..5133)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5793..6584,6585..6653))
                     /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     6416..6435
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6654..6758)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6826..6844
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6882..6904)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7004..7023
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(7104..7120,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(7104..7120,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg tgggattcct gggttaatta
       61 acatcgctag cactaaaatc atagcgccct gtatcaaaaa gaagcttttt tcttttgttg
      121 ggtatgtcta tatagagcac atcctatatt cccttcgagg tgtttttaca agaacaaata
      181 aagaaacaaa ccgcggtgtg aggaagttcc cttaaaaatg gcaaagatta ccgtaaaggt
      241 aaattactcc tttaacgatg cttaaaacga gcatcttact cttgcaacga aattaatcac
      301 tcaatttcaa cgaggatgtt ggcttcccat caaagtagtt ttggggtaaa ccgcatatat
      361 ctgtatatta agaaaactta ttttccacta tctaatttga actacagaga agataagaaa
      421 tcttttaagt agcaatcatg ttttatctaa gtgacatcga ggaagaagcc tcggctgggg
      481 ctgaaccaac ttataatttc tgggaagttt tgttattctc caatactcaa gaaaacctcg
      541 tcaccgttgt cggagagttg cacacactta ccgatcgtgt ggttcattac aaaattgagc
      601 ctgaatcaag ggaagttact gccacaacac tgccatcttt gttagcactt ttgctagaaa
      661 aaaggaacca ggcgagacgt ttatataggg atgtgctttc aatgaaaatg tcagaactgg
      721 actgggacat cgacgatttg ttcacacagt tacaagaaga gctaacaaga actgatgaca
      781 ctctttctat gtatcctagg agaaggttct atcactagct tgctcaacgc gaccttatac
      841 acagagttta atgtgaagag aacattcata tacaattata taaacattgt tgtattctcg
      901 tacatcgtta cacaaaatac gaagcgggga aatagtgata ttcgcttcgc tggtggcaaa
      961 tgggaaaaat acaaaatttt acacatcttt cttccaaata cttccataaa acatctaaat
     1021 atagtggttt ctaattatct ctgatcataa ggaagtggaa gtacatcaga aatgtctcaa
     1081 gaacagtaca cagaaaactt gaaggttatc gttgccgaaa aactggctgg tataccaaac
     1141 tttaacgaag atatcaagta tgttgcggag tatattgtct tattgatcgt taacggtggt
     1201 actgttgaat ctgtcgtaga cgagctggct agtttgtttg atagtgtttc gagagatacg
     1261 cttgcaaatg ttgttcaaac agcctttttc gcattagaag ctctgcaaca gggagaaagt
     1321 gctgaaaata ttgtttccaa aattagaatg atgaatgcgc aaagcttggg acaatcggat
     1381 atcgcacaac agcaacaaca gcaacaacaa caacagcaac cagacatcgc gcaacagcaa
     1441 cctcaacagc aacctcaaca gcaacctcaa cagcaacctc aacagcaacc tcaacagcaa
     1501 cctcaacagc aacctcaaca gcaacctcaa cagcaacctc aacttcaacc acttcagcca
     1561 caactaggga cccagaatgc aatgcagaca gatgctcctg caactccatc ccccatatca
     1621 gccttttccg gcgttgttaa cgctgcagct ccccctcagt ttgcgcctgt agataacagc
     1681 caaaggttca ctcaacgtgg cggaggcgcc gttggaaaga atcgtagagg tggtcgcggt
     1741 gggaaccgtg gaggacgcaa caataattcc acacgtttta atccgttagc aaaagcactt
     1801 ggaatggcgg gtgagagtaa tatgaacttc actccaacca agaaagaggg gcgttgcaga
     1861 ttgtttcctc actgtcctct tggtagatca tgcccacatg cacacccaac taaggtatgt
     1921 aatgaatatc caaattgtcc aaagcctccc ggaacttgtg agtttttaca tccaaatgaa
     1981 gatgaagagt tgatgaagga aatggaaaga actcgtgaag aatttcaaaa aagaaaagct
     2041 gatttattgg cggcaaaaag gaaaccggta caaactggta tcgttctgtg taaatttggg
     2101 gctctgtgtt ccaatccatc atgcccattt ggtcatccaa caccagcaaa tgaagatgcg
     2161 aaagtcattg atctaatgtg gtgtgacaag aatttgacat gtgataatcc tgagtgtaga
     2221 aaggcccact cttcattgtc gaagatcaag gaagtaaaac caataagcca gaagaaagca
     2281 gctccacctc cggttgaaaa gtccttagaa caatgtaagt tcggtacgca ctgcaccaat
     2341 aaacgttgca aatatagaca tgctcgttct catattatgt gccgtgaagg agcaaactgt
     2401 actagaattg attgtttatt tggccatcca attaatgaag attgtagatt tggtgtcaat
     2461 tgtaagaata tttactgtct attcagacat cctccaggca gagtacttcc ggaaaagaaa
     2521 ggcgctgcac ccaattcaaa cgttcctacc aatgaaaggc catttgcatt gccagaaaac
     2581 gcaataattg aaaatgctcc tccgcaaacc agttttacgc accaagaaca agatacggaa
     2641 atgaaccttt ctcttgaagt tgctgaagaa atgagcgaac tgatcaaaga gaacatgcac
     2701 atgaagctgt acatggaagg caccgttgac aaccaccact ttaagtgcac ctctgagggt
     2761 gagggtaagc cctacgaagg cacccaaacc atgcgtatca aagttgtgga gggcggtcca
     2821 ctgccattcg cttttgacat tctggccacc agcttcctgt acggttccaa aaccttcatt
     2881 aaccatactc agggcattcc tgatttcttc aaacagagct ttccggaagg tttcacctgg
     2941 gagcgtgtca ccacgtatga agatggtggt gtgttgaccg ccacccaaga tacctccctg
     3001 caagatggct gtctgatcta taacgtgaaa attcgtggcg tcaactttac tagcaatggt
     3061 cccgtgatgc agaagaaaac cctgggttgg gaggccttta cagaaaccct gtatcctgcc
     3121 gatggtggcc tggagggccg taacgacatg gcactgaagc tggttggtgg cagccatttg
     3181 atcgcaaatg ccaagaccac ttacagaagc aagaaacccg caaaaaatct gaagatgccg
     3241 ggtgtttact atgtcgacta ccgtctggaa agaattaaag aagcaaataa tgagacttac
     3301 gtggagcagc acgaggttgc agtcgcgaga tattgcgact tgcctagcaa gctgggtcat
     3361 aaactgaatt gaggcgcgcc acttctaaat aagcgaattt cttatgattt atgattttta
     3421 ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt
     3481 taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg
     3541 tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct agggataaca
     3601 gggtaatatc tgcaggtcga caacccttaa tataacttcg tataatgtat gctatacgaa
     3661 gttattaggt ctagagcggc cgcggaacga tcattcacta tatatatatc aatttatata
     3721 tacgtatgtg taattgaaga aagatacgtt ttttcctcta ttgagaggcc tgctggatga
     3781 atagctttac cttttctaaa tccttgatac catcagtctc tactcctcca cttacatcga
     3841 caccgatcgc atttggtaac atattaatgg caacagaaac gttatcagga ttcaatccac
     3901 cagcgataat gaattttatc tcgggatgac ttgcagacca actggaaatt gcactccaat
     3961 tcaatttctc accagtgcca ccttcaccag aatcgaacaa cgtcagcaca ttgtctacgt
     4021 gttcatacag gtccagtagt aattcacaat cctgtgggaa ctggaacctc ttaatgattg
     4081 gaattgaaga tgggatcaaa gatctgtatt ctttaatatc ttcatctcca tgtaattgta
     4141 tcacatctag attatattcg tggtacagtt gaaggacatc atcaacggac tgatttctaa
     4201 acaccccgac tagtttagta cctttcacgt tctcttgttg gtgaactgca gttgaaatac
     4261 ctttcgcaac agatgagtca atggttcttt tcctaccggg aacacaaatg atacctaagt
     4321 aatcagcacc atcatccaca gcagtctttg cagcttcaac ggtttgcaaa ccacacactt
     4381 taacgagcat cacagtagat ttcgcaagag ggtagctcgc tcagagtacc caagtaaatg
     4441 attagtaaac tgatgtttga tagttcaatt tttcaatgaa ataaccttat attaaaattg
     4501 atattactat tatacaaaaa taaagaataa aggatttgag tttatacata aaataccatt
     4561 attatttgtt cagtgagaga taccggggta tatgggatgt gtgtagtgat accatgcaat
     4621 catgtatcaa acatgggccc ggtaccgagc tctggtcgag aacccttaat ataacttcgt
     4681 ataatgtatg ctatacgaag ttattaggtg atatcagatc cactagtggc ctatgcggcc
     4741 gcggatctgc cggtctccct atagtgagtc gtattaattt cgataagcca ggttaacctg
     4801 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4861 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4921 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4981 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5041 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5101 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5161 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5221 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5281 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5341 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5401 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5461 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5521 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5581 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5641 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5701 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5761 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5821 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5881 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5941 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6001 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6061 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6121 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6181 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6241 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6301 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6361 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6421 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6481 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6541 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6601 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6661 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6721 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6781 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     6841 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     6901 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     6961 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     7021 gtactgagag tgcaccatat ggacatattg tcgttagaac gcggctacaa ttaatacata
     7081 accttatgta tcatacacat acgatttagg tgacactata
//
