LOCUS       sequence-256864-        7129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of NAB2-mCherry 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 7129)
  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 7129)
  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: ZJOM82
FEATURES             Location/Qualifiers
     source          1..7129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2671..3381
                     /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(2816..2834)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3084..3103
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      3402..3589
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3457..3481)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    3641..3674
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4680..4713
                     /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(4766..4785)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4767..4785)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4872..4889)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5043..5631)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5123..5142)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5802..6593,6594..6662))
                     /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     6425..6444
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6663..6767)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6835..6853
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6891..6913)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7013..7032
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(7113..7129,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(7113..7129,1)
                     /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 atggtgagca agggcgagga ggataacatg
     2701 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     2761 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     2821 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     2881 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
     2941 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     3001 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     3061 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     3121 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     3181 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     3241 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
     3301 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     3361 atggacgagc tgtacaagtg aggcgcgcca cttctaaata agcgaatttc ttatgattta
     3421 tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact
     3481 cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc
     3541 tttctcaggt atagtatgag gtcgctctta ttgaccacac ctctaccggc agatccgcta
     3601 gggataacag ggtaatatct gcaggtcgac aacccttaat ataacttcgt ataatgtatg
     3661 ctatacgaag ttattaggtc tagagcggcc gcggaacgat cattcactat atatatatca
     3721 atttatatat acgtatgtgt aattgaagaa agatacgttt tttcctctat tgagaggcct
     3781 gctggatgaa tagctttacc ttttctaaat ccttgatacc atcagtctct actcctccac
     3841 ttacatcgac accgatcgca tttggtaaca tattaatggc aacagaaacg ttatcaggat
     3901 tcaatccacc agcgataatg aattttatct cgggatgact tgcagaccaa ctggaaattg
     3961 cactccaatt caatttctca ccagtgccac cttcaccaga atcgaacaac gtcagcacat
     4021 tgtctacgtg ttcatacagg tccagtagta attcacaatc ctgtgggaac tggaacctct
     4081 taatgattgg aattgaagat gggatcaaag atctgtattc tttaatatct tcatctccat
     4141 gtaattgtat cacatctaga ttatattcgt ggtacagttg aaggacatca tcaacggact
     4201 gatttctaaa caccccgact agtttagtac ctttcacgtt ctcttgttgg tgaactgcag
     4261 ttgaaatacc tttcgcaaca gatgagtcaa tggttctttt cctaccggga acacaaatga
     4321 tacctaagta atcagcacca tcatccacag cagtctttgc agcttcaacg gtttgcaaac
     4381 cacacacttt aacgagcatc acagtagatt tcgcaagagg gtagctcgct cagagtaccc
     4441 aagtaaatga ttagtaaact gatgtttgat agttcaattt ttcaatgaaa taaccttata
     4501 ttaaaattga tattactatt atacaaaaat aaagaataaa ggatttgagt ttatacataa
     4561 aataccatta ttatttgttc agtgagagat accggggtat atgggatgtg tgtagtgata
     4621 ccatgcaatc atgtatcaaa catgggcccg gtaccgagct ctggtcgaga acccttaata
     4681 taacttcgta taatgtatgc tatacgaagt tattaggtga tatcagatcc actagtggcc
     4741 tatgcggccg cggatctgcc ggtctcccta tagtgagtcg tattaatttc gataagccag
     4801 gttaacctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4861 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4921 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4981 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5041 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5101 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5161 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5221 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5281 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5341 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5401 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5461 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     5521 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5581 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5641 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5701 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5761 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5821 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5881 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5941 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6001 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6061 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6121 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6181 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6241 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6301 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6361 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6421 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6481 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6541 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6601 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6661 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6721 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6781 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     6841 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     6901 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     6961 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     7021 gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg cggctacaat
     7081 taatacataa ccttatgtat catacacata cgatttaggt gacactata
//
