LOCUS       sequence-269551-        7126 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 7126)
  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 7126)
  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..7126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2640..3350
                     /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(2785..2803)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3053..3072
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      3371..3558
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3426..3450)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    3610..3643
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4646..4679
                     /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(4732..4751)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4733..4751)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4838..4855)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5009..5597)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5089..5108)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5768..6559,6560..6628))
                     /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     6391..6410
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6629..6733)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6801..6819
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6857..6879)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6979..6998
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7079..7097
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7079..7096
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 tgcaggtcgt gggattcctg ggttaattaa catcgctagc actaaaatca tagcgccctg
       61 tatcaaaaag aagctttttt cttttgttgg gtatgtctat atagagcaca tcctatattc
      121 ccttcgaggt gtttttacaa gaacaaataa agaaacaaac cgcggtgtga ggaagttccc
      181 ttaaaaatgg caaagattac cgtaaaggta aattactcct ttaacgatgc ttaaaacgag
      241 catcttactc ttgcaacgaa attaatcact caatttcaac gaggatgttg gcttcccatc
      301 aaagtagttt tggggtaaac cgcatatatc tgtatattaa gaaaacttat tttccactat
      361 ctaatttgaa ctacagagaa gataagaaat cttttaagta gcaatcatgt tttatctaag
      421 tgacatcgag gaagaagcct cggctggggc tgaaccaact tataatttct gggaagtttt
      481 gttattctcc aatactcaag aaaacctcgt caccgttgtc ggagagttgc acacacttac
      541 cgatcgtgtg gttcattaca aaattgagcc tgaatcaagg gaagttactg ccacaacact
      601 gccatctttg ttagcacttt tgctagaaaa aaggaaccag gcgagacgtt tatataggga
      661 tgtgctttca atgaaaatgt cagaactgga ctgggacatc gacgatttgt tcacacagtt
      721 acaagaagag ctaacaagaa ctgatgacac tctttctatg tatcctagga gaaggttcta
      781 tcactagctt gctcaacgcg accttataca cagagtttaa tgtgaagaga accttcatat
      841 acaattatat aaacattgtt gtattctcgt acatcgttac acaaaatacg aagcggggaa
      901 atagtgatat tcgcttcgct ggtggcaaat gggaaaaata caaaatttta cacatctttc
      961 ttccaaatac ttccataaaa catctaaata tagtggtttc taattatctc tgatcataag
     1021 gaagtggaag tacatcagaa atgtctcaag aacagtacac agaaaacttg aaggttatcg
     1081 ttgccgaaaa actggctggt ataccaaact ttaacgaaga tatcaagtat gttgcggagt
     1141 atattgtctt attgatcgtt aacggtggta ctgttgaatc tgtcgtagac gagctggcta
     1201 gtttgtttga tagtgtttcg agagatacgc ttgcaaatgt tgttcaaaca gcctttttcg
     1261 cattagaagc tctgcaacag ggagaaagtg ctgaaaatat tgtttccaaa attagaatga
     1321 tgaatgcgca aagcttggga caatcggata tcgcacaaca gcaacaacag caacaacaac
     1381 aacagcaacc agacatcgcg caacagcaac ctcaacagca acctcaacag caacctcaac
     1441 agcaacctca acagcaacct caacagcaac ctcaacagca acctcaacag caacctcaac
     1501 agcaacctca acttcaacca cttcagccac aactagggac ccagaatgca atgcagacag
     1561 atgctcctgc aactccatcc cccatatcag ccttttccgg cgttgttaac gctgcagctc
     1621 cccctcagtt tgcgcctgta gataacagcc aaaggttcac tcaacgtggc ggaggcgccg
     1681 ttggaaagaa tcgtagaggt ggtcgcggtg ggaaccgtgg aggacgcaac aataattcca
     1741 cacgttttaa tccgttagca aaagcacttg gaatggcggg tgagagtaat atgaacttca
     1801 ctccaaccaa gaaagagggg cgttgcagat tgtttcctca ctgtcctctt ggtagatcat
     1861 gcccacatgc acacccaact aaggtatgta atgaatatcc aaattgtcca aagcctcccg
     1921 gaacttgtga gtttttacat ccaaatgaag atgaagagtt gatgaaggaa atggaaagaa
     1981 ctcgtgaaga atttcaaaaa agaaaagctg atttattggc ggcaaaaagg aaaccggtac
     2041 aaactggtat cgttctgtgt aaatttgggg ctctgtgttc caatccatca tgcccatttg
     2101 gtcatccaac accagcaaat gaagatgcga aagtcattga tctaatgtgg tgtgacaaga
     2161 atttgacatg tgataatcct gagtgtagaa aggcccactc ttcattgtcg aagatcaagg
     2221 aagtaaaacc aataagccag aagaaagcag ctccacctcc ggttgaaaag tccttagaac
     2281 aatgtaagtt cggtacgcac tgcaccaata aacgttgcaa atatagacat gctcgttctc
     2341 atattatgtg ccgtgaagga gcaaactgta ctagaattga ttgtttattt ggccatccaa
     2401 ttaatgaaga ttgtagattt ggtgtcaatt gtaagaatat ttactgtcta ttcagacatc
     2461 ctccaggcag agtacttccg gaaaagaaag gcgctgcacc caattcaaac gttcctacca
     2521 atgaaaggcc atttgcattg ccagaaaacg caataattga aaatgctcct ccgcaaacca
     2581 gttttacgca ccaagaacaa gatacggaaa tgaacctttc tcttgaagtt gctgaagaaa
     2641 tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
     2701 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     2761 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
     2821 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
     2881 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
     2941 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
     3001 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
     3061 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
     3121 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
     3181 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
     3241 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     3301 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtga ggcgcgccac
     3361 ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa
     3421 aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt
     3481 gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg tcgctcttat
     3541 tgaccacacc tctaccggca gatccgctag ggataacagg gtaatatctg caggtcgaca
     3601 acccttaata taacttcgta taatgtatgc tatacgaagt tattaggtct agagcggccg
     3661 cggaacgatc attcactata tatatatcaa tttatatata cgtatgtgta attgaagaaa
     3721 gatacgtttt ttcctctatt gagaggcctg ctggatgaat agctttacct tttctaaatc
     3781 cttgatacca tcagtctcta ctcctccact tacatcgaca ccgatcgcat ttggtaacat
     3841 attaatggca acagaaacgt tatcaggatt caatccacca gcgataatga attttatctc
     3901 gggatgactt gcagaccaac tggaaattgc actccaattc aatttctcac cagtgccacc
     3961 ttcaccagaa tcgaacaacg tcagcacatt gtctacgtgt tcatacaggt ccagtagtaa
     4021 ttcacaatcc tgtgggaact ggaacctctt aatgattgga attgaagatg ggatcaaaga
     4081 tctgtattct ttaatatctt catctccatg taattgtatc acatctagat tatattcgtg
     4141 gtacagttga aggacatcat caacggactg atttctaaac accccgacta gtttagtacc
     4201 tttcacgttc tcttgttggt gaactgcagt tgaaatacct ttcgcaacag atgagtcaat
     4261 ggttcttttc ctaccgggaa cacaaatgat acctaagtaa tcagcaccat catccacagc
     4321 agtctttgca gcttcaacgg tttgcaaacc acacacttta acgagcatca cagtagattt
     4381 cgcaagaggg tagctcgctc agagtaccca agtaaatgat tagtaaactg atgtttgata
     4441 gttcaatttt tcaatgaaat aaccttatat taaaattgat attactatta tacaaaaata
     4501 aagaataaag gatttgagtt tatacataaa ataccattat tatttgttca gtgagagata
     4561 ctggggtata tgggatgtgt gtagtgatac catgcaatca tgtatcaaac atgggcccgg
     4621 taccgagctc tcgagaaccc ttaatataac ttcgtataat gtatgctata cgaagttatt
     4681 aggtgatatc agatccacta gtggcctatg cggccgcgga tctgccggtc tccctatagt
     4741 gagtcgtatt aatttcgata agccaggtta acctgcatta atgaatcggc caacgcgcgg
     4801 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4861 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4921 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4981 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5041 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5101 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5161 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5221 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5281 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5341 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5401 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5461 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5521 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5581 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5641 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5701 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5761 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5821 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5881 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5941 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6001 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6061 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6121 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6181 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6241 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6301 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6361 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6421 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6481 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6541 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6601 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6661 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6721 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     6781 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     6841 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     6901 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     6961 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatggaca
     7021 tattgtcgtt agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat
     7081 ttaggtgaca ctatagaacg cggccgccag ctgaagcttc gtacgc
//
