LOCUS       sequence-267445-        7051 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of ANP1-mCherry as an additional copy in genome, use 
            auxotrophic marker TRP1(Kluyveromyces lactis). Marker for yeast 
            early golgi..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7051)
  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 7051)
  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: ZJOM75
FEATURES             Location/Qualifiers
     source          1..7051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             2613..3323
                     /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(2758..2776)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3026..3045
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      3344..3531
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3399..3423)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    3583..3616
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4619..4652
                     /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(4705..4724)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4706..4724)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4811..4828)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4982..5570)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5062..5081)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5741..6532,6533..6601))
                     /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     6364..6383
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6602..6706)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6774..6792
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6830..6852)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6952..6971
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaggtga cactatagaa cgcggccgcc agctgaagct tcgtacgctg caggtcgtgg
       61 gattcctggg ttaattaaca tcgctagcga ttccaatgtc aagatatggc aaaaaccttg
      121 tgcattatat aatcgtggaa cacgacgatc aacgcggcca aaagccgatc gacgacgacg
      181 acgaaaaaaa tttttattac cactgttcct tcaccttcga aacttttttt cgggcgacgg
      241 cgtttctttt agctcctgcg gtatgtgcgg tgcgggaggt cccttgtcgc ctcacacgga
      301 cccgatataa cgctacggaa tacatcgagg ggtatgggtg gatgatatca ctgcagcaag
      361 gtctgggtgt tgcagaattt taccggcctt ggcccctcag cgtccagcta cagcgttata
      421 ctactccaag tcgccgttcc cgttttgctg ttctcacccc tcagagaaaa tgccaccaga
      481 acgaagccaa tggtcaggac ttatcgttac ttatactgtt gagccggata tatccgctgt
      541 gctctaatac cagccaaacg agaagggtag cggcaagaaa gggtaaactt tcatgatgaa
      601 gttagctagt tccttttacg attgctacta tctcgttcaa cttggcctgc gctgttgatg
      661 aaaatagcag tgactgcaga gaatagaaat gaaccgggct catctggtct agttgccttc
      721 ttttttttat tttttttcat tttccttttc ttttcttttc ttttcttgtc tcactatgtt
      781 ggtttaatca attcgtaatc cctttaccaa cgcattgtgt tcaattttcc tcttgtgtgg
      841 ggtcgcccgc cttgtctcat cgtgaaaaaa ttgcgtaaga tgaaatcatg aaaacatgaa
      901 ataaaaaaaa taaataaaaa taactacacg attcaaaaac gccaagatct gttagtagag
      961 acagcaagca ttgcaaaacc attcatcact cagttgctag gcgagtgttg gtgcgattta
     1021 ccctcatcat ctttttttcg ctccattaaa cagttccttg tggctttcat gcaaagtgta
     1081 cgtccataat gaagtataat aacagaaaac tctcgttcaa ccctaccaca gtaagtatcg
     1141 ctggaacgtt gcttacggtg ttctttctca caagactcgt gctttcgttc ttctcgatat
     1201 cgctattcca gctggtaact ttccaaggaa tcttcaagcc ctatgttcca gattttaaaa
     1261 atactcccag cgtagagttc tacgacctac gaaattacca aggcaacaaa gatggttggc
     1321 aacagggtga ccgcatcttg ttttgcgtgc cactgagaga tgcttctgag catcttccca
     1381 tgttttttaa ccatctgaat accatgacat atccgcacaa cctgattgat ctgtcgtttt
     1441 tggtgagtga ctcttcggat aacaccatgg gcgtgttact atccaatcta cagatggcgc
     1501 aatcgcaaca agacaagtcc aaaaggtttg gtaacatcga aatctatgag aaggacttcg
     1561 gccaaatcat tggtcagtcc ttttctgatc gtcacgggtt tggcgcccag ggccccagaa
     1621 gaaagctgat ggccagggct cgtaattggc ttgggtccgt ggccctgaaa ccataccact
     1681 cctgggttta ctggagagac gtggatgtcg aaaccatccc cactaccatc atggaggatc
     1741 taatgcatca cgacaaggac gtcattgtcc ccaacgtctg gagaccttta ccagactggc
     1801 tgggaaacat tcaaccctac gatttaaatt catggaagga atcagaaggg ggtcttcaat
     1861 tggcggattc cctagacgaa gatgctgtca ttgtggaagg gtacccagag tacgccactt
     1921 ggagacctca cctggcatac atgagagacc caaacggtaa tccagaagac gaaatggaat
     1981 tggacggtat tggaggggtc tcgattctcg cgaaagctaa agtattcaga acaggctcac
     2041 actttcctgc attctccttt gaaaagcacg cagagacaga agcattcggt aggctctccc
     2101 gcaggatgaa ttacaatgtc atcggcttgc cccattatgt catctggcac atctacgaac
     2161 cttccagcga cgacttaaag cacatggctt ggatggctga agaggagaag cggaaactag
     2221 aagaagagag aattcgcgaa ttttacaaca agatctggga gattggattt gaagatgtca
     2281 gagaccaatg gaacgaagaa agagattcaa tcttaaaaaa cattgactcc actttgaaca
     2341 ataaagtcac tgtagactgg tctgaagagg gagatggctc ggagctggtg gactcaaagg
     2401 gtgactttgt ctcaccaaac aaccaacaac aacaacaaca gcaacaacaa caacaacagc
     2461 agcagcagca acagcaacag caacaacagc tcgacggcaa cccgcagggg aaacctcttg
     2521 atgacaacga caagaacaag aaaaaacatc ctaaagaagt tccattagac ttcgaccctg
     2581 atagaaacct ttctcttgaa gttgctgaag aaatggtgag caagggcgag gaggataaca
     2641 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
     2701 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     2761 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
     2821 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
     2881 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     2941 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     3001 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     3061 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
     3121 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     3181 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     3241 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     3301 gcatggacga gctgtacaag tgaggcgcgc cacttctaaa taagcgaatt tcttatgatt
     3361 tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga
     3421 ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt
     3481 gctttctcag gtatagtatg aggtcgctct tattgaccac acctctaccg gcagatccgc
     3541 tagggataac agggtaatat ctgcaggtcg acaaccctta atataacttc gtataatgta
     3601 tgctatacga agttattagg tctagagcgg ccgcggaacg atcattcact atatatatat
     3661 caatttatat atacgtatgt gtaattgaag aaagatacgt tttttcctct attgagaggc
     3721 ctgctggatg aatagcttta ccttttctaa atccttgata ccatcagtct ctactcctcc
     3781 acttacatcg acaccgatcg catttggtaa catattaatg gcaacagaaa cgttatcagg
     3841 attcaatcca ccagcgataa tgaattttat ctcgggatga cttgcagacc aactggaaat
     3901 tgcactccaa ttcaatttct caccagtgcc accttcacca gaatcgaaca acgtcagcac
     3961 attgtctacg tgttcataca ggtccagtag taattcacaa tcctgtggga actggaacct
     4021 cttaatgatt ggaattgaag atgggatcaa agatctgtat tctttaatat cttcatctcc
     4081 atgtaattgt atcacatcta gattatattc gtggtacagt tgaaggacat catcaacgga
     4141 ctgatttcta aacaccccga ctagtttagt acctttcacg ttctcttgtt ggtgaactgc
     4201 agttgaaata cctttcgcaa cagatgagtc aatggttctt ttcctaccgg gaacacaaat
     4261 gatacctaag taatcagcac catcatccac agcagtcttt gcagcttcaa cggtttgcaa
     4321 accacacact ttaacgagca tcacagtaga tttcgcaaga gggtagctcg ctcagagtac
     4381 ccaagtaaat gattagtaaa ctgatgtttg atagttcaat ttttcaatga aataacctta
     4441 tattaaaatt gatattacta ttatacaaaa ataaagaata aaggatttga gtttatacat
     4501 aaaataccat tattatttgt tcagtgagag atactggggt atatgggatg tgtgtagtga
     4561 taccatgcaa tcatgtatca aacatgggcc cggtaccgag ctctcgagaa cccttaatat
     4621 aacttcgtat aatgtatgct atacgaagtt attaggtgat atcagatcca ctagtggcct
     4681 atgcggccgc ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg
     4741 ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4801 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4861 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4921 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4981 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5041 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5101 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5161 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5221 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5281 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5341 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5401 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5461 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5521 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5581 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5641 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5701 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5761 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5821 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5881 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5941 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6001 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6061 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6121 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6181 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6241 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6301 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6361 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6421 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6481 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6541 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6601 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6661 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6721 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     6781 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     6841 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     6901 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     6961 agcagattgt actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt
     7021 aatacataac cttatgtatc atacacatac g
//
