LOCUS       sequence-265903-        6260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of SNF7-mCherry as an additional copy in genome, use 
            auxotrophic marker TRP1(Kluyveromyces lactis). Marker for yeast late
            endosome..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6260)
  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 6260)
  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: ZJOM44
FEATURES             Location/Qualifiers
     source          1..6260
                     /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             1813..2523
                     /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(1958..1976)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2226..2245
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      2553..2740
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2608..2632)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    2792..2825
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3828..3861
                     /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(3914..3933)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3915..3933)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4020..4037)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4191..4779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4271..4290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4950..5741,5742..5810))
                     /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     5573..5592
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5811..5915)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5983..6001
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6039..6061)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6161..6180
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaggtga cactatagaa cgcggccgcc agctgaagct tatgcgttga ttattgggtt
       61 tctccccaca gctgtacatt gacgatgctc tttattgata tcgcctaaaa gttcaacaaa
      121 cgccactgga aattgccgcc gttgtcacca caacgtgata agtgatggtg tgctttttgc
      181 acatctagtg aagaaacggg caattaggat ctaggaccag taacacttgc cccgcaaata
      241 ttataacatg ggccttatga ttgctcggtt tgttacatct aacatcctaa gaagtgagtg
      301 gcatgaatac gtagctatat gtagattgaa aattgatgag tcgagtcatc accttcctct
      361 aaacggggga taaattttgg cggtattttc cctaatttta ttcttcgtta tgtcagccat
      421 gtgagactaa ttaattcaag ttttgactta caattgcggc ttcgtcggaa atcagtatta
      481 cgtataaact gtgtaccata aaattggcgt actcggtatt gtttatacta agccggccat
      541 tctagtgatt tcgcctctat aaatggttgt tattcgagta taatatttca accgcatcaa
      601 agaaagaggt agatagcttg cgttcaacac cgttcttttc tttccccctt aatttagcgc
      661 ttggcggtta tgctatggtg acagttaact tttttaatta taggatgact tgtcatattg
      721 ttatatgccg catacttatg taaagtagtt attgaggtgg gggtgagcag agtaacgaca
      781 acgatggagt tccgtaacta accaatgctt aatgttggta agtaatagca aatatatata
      841 ctacatagct tttgttatat gcatcatttt atacgattca tgttcgaccc ttttgataaa
      901 agatgggatt tttaaataag tcggtgggtg atttcgaatc atgaatagtt ggcagagcag
      961 gggaagtacg agcttctaaa gggtaagata ttgtatttcg gacggaagca gcagaaacat
     1021 aacagtattg ataaataagg catgtggtca tcactttttg gttggacatc aagtaatgcc
     1081 aagaataaag agtcaccaac aaaggccata gtgcggttga gggagcatat caaccttcta
     1141 tccaaaaagc aatcgcattt acgtactcaa attacaaatc aagagaatga agctagaatc
     1201 tttttgacga agggcaataa agtaatggcg aagaatgcac ttaaaaagaa gaagacaatc
     1261 gaacaacttt taagtaaggt agaaggcaca atggagtcta tggaacagca gctattctct
     1321 atagaaagtg caaacttaaa tctagagaca atgagggcta tgcaggaagg tgctaaggca
     1381 atgaaaacta ttcacagtgg ccttgacata gataaagtgg atgaaactat ggacgagata
     1441 agggagcaag tcgaattagg agatgaaata agcgacgcta tatccaggcc tttaattact
     1501 ggggcaaacg aggtggatga agatgagctg gacgaggaat tggacatgct ggctcaagaa
     1561 aatgctaacc aagaaacgtc caagatcgtt aataataatg ttaatgcggc gcctatctca
     1621 gagaacaaag tctcactacc tagtgttcca agtaataaaa ttaaacaaag tgagaactct
     1681 gtgaaggacg gggaagagga agaggatgaa gaagatgaag atgaaaaagc attaagagaa
     1741 ctacaagcag aaatggggct tgcttcgtac gctgcaggtc gtgggattcc tgggttaatt
     1801 aacatcgcta gcatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     1861 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     1921 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     1981 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     2041 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     2101 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     2161 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     2221 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     2281 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     2341 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     2401 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     2461 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     2521 tagcctaggt gaggcgcgcc acttctaaat aagcgaattt cttatgattt atgattttta
     2581 ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac tcttaggttt
     2641 taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg ctttctcagg
     2701 tatagtatga ggtcgctctt attgaccaca cctctaccgg cagatccgct agggataaca
     2761 gggtaatatc tgcaggtcga caacccttaa tataacttcg tataatgtat gctatacgaa
     2821 gttattaggt ctagagcggc cgcggaacga tcattcacta tatatatatc aatttatata
     2881 tacgtatgtg taattgaaga aagatacgtt ttttcctcta ttgagaggcc tgctggatga
     2941 atagctttac cttttctaaa tccttgatac catcagtctc tactcctcca cttacatcga
     3001 caccgatcgc atttggtaac atattaatgg caacagaaac gttatcagga ttcaatccac
     3061 cagcgataat gaattttatc tcgggatgac ttgcagacca actggaaatt gcactccaat
     3121 tcaatttctc accagtgcca ccttcaccag aatcgaacaa cgtcagcaca ttgtctacgt
     3181 gttcatacag gtccagtagt aattcacaat cctgtgggaa ctggaacctc ttaatgattg
     3241 gaattgaaga tgggatcaaa gatctgtatt ctttaatatc ttcatctcca tgtaattgta
     3301 tcacatctag attatattcg tggtacagtt gaaggacatc atcaacggac tgatttctaa
     3361 acaccccgac tagtttagta cctttcacgt tctcttgttg gtgaactgca gttgaaatac
     3421 ctttcgcaac agatgagtca atggttcttt tcctaccggg aacacaaatg atacctaagt
     3481 aatcagcacc atcatccaca gcagtctttg cagcttcaac ggtttgcaaa ccacacactt
     3541 taacgagcat cacagtagat ttcgcaagag ggtagctcgc tcagagtacc caagtaaatg
     3601 attagtaaac tgatgtttga tagttcaatt tttcaatgaa ataaccttat attaaaattg
     3661 atattactat tatacaaaaa taaagaataa aggatttgag tttatacata aaataccatt
     3721 attatttgtt cagtgagaga tactggggta tatgggatgt gtgtagtgat accatgcaat
     3781 catgtatcaa acatgggccc ggtaccgagc tctcgagaac ccttaatata acttcgtata
     3841 atgtatgcta tacgaagtta ttaggtgata tcagatccac tagtggccta tgcggccgcg
     3901 gatctgccgg tctccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat
     3961 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4021 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4081 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4141 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4201 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4261 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4321 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4381 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4441 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4501 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4561 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4621 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4681 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4741 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4801 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4861 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4921 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4981 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5041 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5101 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5161 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5221 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5281 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5341 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5401 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5461 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5521 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5581 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5641 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5701 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5761 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5821 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5881 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5941 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     6001 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     6061 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     6121 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     6181 ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta atacataacc
     6241 ttatgtatca tacacatacg
//
