LOCUS       sequence-256865-        7054 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 7054)
  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 7054)
  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..7054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2596..3306
                     /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(2741..2759)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3009..3028
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      3327..3514
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3382..3406)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    3566..3599
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4605..4638
                     /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(4691..4710)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4692..4710)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4797..4814)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4968..5556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5048..5067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5727..6518,6519..6587))
                     /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     6350..6369
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6588..6692)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6760..6778
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6816..6838)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6938..6957
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(7038..7054,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(7038..7054,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg tgggattcct gggttaatta
       61 acatcgctag cgattccaat gtcaagatat ggcaaaaacc ttgtgcatta tataatcgtg
      121 gaacacgacg atcaacgcgg ccaaaagccg atcgacgacg acgacgaaaa aaatttttat
      181 taccactgtt ccttcacctt cgaaactttt tttcgggcga cggcgtttct tttagctcct
      241 gcggtatgtg cggtgcggga ggtcccttgt cgcctcacac ggacccgata taacgctacg
      301 gaatacatcg aggggtatgg gtggatgata tcactgcagc aaggtctggg tgttgcagaa
      361 ttttaccggc cttggcccct cagcgtccag ctacagcgtt atactactcc aagtcgccgt
      421 tcccgttttg ctgttctcac ccctcagaga aaatgccacc agaacgaagc caatggtcag
      481 gacttatcgt tacttatact gttgagccgg atatatccgc tgtgctctaa taccagccaa
      541 acgagaaggg tagcggcaag aaagggtaaa ctttcatgat gaagttagct agttcctttt
      601 acgattgcta ctatctcgtt caacttggcc tgcgctgttg atgaaaatag cagtgactgc
      661 agagaataga aatgaaccgg gctcatctgg tctagttgcc ttcttttttt tatttttttt
      721 cattttcctt ttcttttctt ttcttttctt gtctcactat gttggtttaa tcaattcgta
      781 atccctttac caacgcattg tgttcaattt tcctcttgtg tggggtcgcc cgccttgtct
      841 catcgtgaaa aaattgcgta agatgaaatc atgaaaacat gaaataaaaa aaataaataa
      901 aaataactac acgattcaaa aacgccaaga tctgttagta gagacagcaa gcattgcaaa
      961 accattcatc actcagttgc taggcgagtg ttggtgcgat ttaccctcat catctttttt
     1021 tcgctccatt aaacagttcc ttgtggcttt catgcaaagt gtacgtccat aatgaagtat
     1081 aataacagaa aactctcgtt caaccctacc acagtaagta tcgctggaac gttgcttacg
     1141 gtgttctttc tcacaagact cgtgctttcg ttcttctcga tatcgctatt ccagctggta
     1201 actttccaag gaatcttcaa gccctatgtt ccagatttta aaaatactcc cagcgtagag
     1261 ttctacgacc tacgaaatta ccaaggcaac aaagatggtt ggcaacaggg tgaccgcatc
     1321 ttgttttgcg tgccactgag agatgcttct gagcatcttc ccatgttttt taaccatctg
     1381 aataccatga catatccgca caacctgatt gatctgtcgt ttttggtgag tgactcttcg
     1441 gataacacca tgggcgtgtt actatccaat ctacagatgg cgcaatcgca acaagacaag
     1501 tccaaaaggt ttggtaacat cgaaatctat gagaaggact tcggccaaat cattggtcag
     1561 tccttttctg atcgtcacgg gtttggcgcc cagggcccca gaagaaagct gatggccagg
     1621 gctcgtaatt ggcttgggtc cgtggccctg aaaccatacc actcctgggt ttactggaga
     1681 gacgtggatg tcgaaaccat ccccactacc atcatggagg atctaatgca tcacgacaag
     1741 gacgtcattg tccccaacgt ctggagacct ttaccagact ggctgggaaa cattcaaccc
     1801 tacgatttaa attcatggaa ggaatcagaa gggggtcttc aattggcgga ttccctagac
     1861 gaagatgctg tcattgtgga agggtaccca gagtacgcca cttggagacc tcacctggca
     1921 tacatgagag acccaaacgg taatccagaa gacgaaatgg aattggacgg tattggaggg
     1981 gtctcgattc tcgcgaaagc taaagtattc agaacaggct cacactttcc tgcattctcc
     2041 tttgaaaagc acgcagagac agaagcattc ggtaggctct cccgcaggat gaattacaat
     2101 gtcatcggct tgccccatta tgtcatctgg cacatctacg aaccttccag cgacgactta
     2161 aagcacatgg cttggatggc tgaagaggag aagcggaaac tagaagaaga gagaattcgc
     2221 gaattttaca acaagatctg ggagattgga tttgaagatg tcagagacca atggaacgaa
     2281 gaaagagatt caatcttaaa aaacattgac tccactttga acaataaagt cactgtagac
     2341 tggtctgaag agggagatgg ctcggagctg gtggactcaa agggtgactt tgtctcacca
     2401 aacaaccaac aacaacaaca acagcaacaa caacaacaac agcagcagca gcaacagcaa
     2461 cagcaacaac agctcgacgg caacccgcag gggaaacctc ttgatgacaa cgacaagaac
     2521 aagaaaaaac atcctaaaga agttccatta gacttcgacc ctgatagaaa cctttctctt
     2581 gaagttgctg aagaaatggt gagcaagggc gaggaggata acatggccat catcaaggag
     2641 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag
     2701 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     2761 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     2821 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
     2881 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     2941 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
     3001 tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg
     3061 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
     3121 ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     3181 ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc
     3241 gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     3301 aagtgaggcg cgccacttct aaataagcga atttcttatg atttatgatt tttattatta
     3361 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
     3421 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt
     3481 atgaggtcgc tcttattgac cacacctcta ccggcagatc cgctagggat aacagggtaa
     3541 tatctgcagg tcgacaaccc ttaatataac ttcgtataat gtatgctata cgaagttatt
     3601 aggtctagag cggccgcgga acgatcattc actatatata tatcaattta tatatacgta
     3661 tgtgtaattg aagaaagata cgttttttcc tctattgaga ggcctgctgg atgaatagct
     3721 ttaccttttc taaatccttg ataccatcag tctctactcc tccacttaca tcgacaccga
     3781 tcgcatttgg taacatatta atggcaacag aaacgttatc aggattcaat ccaccagcga
     3841 taatgaattt tatctcggga tgacttgcag accaactgga aattgcactc caattcaatt
     3901 tctcaccagt gccaccttca ccagaatcga acaacgtcag cacattgtct acgtgttcat
     3961 acaggtccag tagtaattca caatcctgtg ggaactggaa cctcttaatg attggaattg
     4021 aagatgggat caaagatctg tattctttaa tatcttcatc tccatgtaat tgtatcacat
     4081 ctagattata ttcgtggtac agttgaagga catcatcaac ggactgattt ctaaacaccc
     4141 cgactagttt agtacctttc acgttctctt gttggtgaac tgcagttgaa atacctttcg
     4201 caacagatga gtcaatggtt cttttcctac cgggaacaca aatgatacct aagtaatcag
     4261 caccatcatc cacagcagtc tttgcagctt caacggtttg caaaccacac actttaacga
     4321 gcatcacagt agatttcgca agagggtagc tcgctcagag tacccaagta aatgattagt
     4381 aaactgatgt ttgatagttc aatttttcaa tgaaataacc ttatattaaa attgatatta
     4441 ctattataca aaaataaaga ataaaggatt tgagtttata cataaaatac cattattatt
     4501 tgttcagtga gagataccgg ggtatatggg atgtgtgtag tgataccatg caatcatgta
     4561 tcaaacatgg gcccggtacc gagctctggt cgagaaccct taatataact tcgtataatg
     4621 tatgctatac gaagttatta ggtgatatca gatccactag tggcctatgc ggccgcggat
     4681 ctgccggtct ccctatagtg agtcgtatta atttcgataa gccaggttaa cctgcattaa
     4741 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4801 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4861 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4921 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4981 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5041 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5101 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5161 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5221 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5281 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5341 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5401 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5461 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5521 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5581 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5641 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5701 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5761 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5821 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5881 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5941 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6001 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6061 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6121 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6181 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6241 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6301 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6361 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6421 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6481 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6541 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6601 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6661 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6721 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     6781 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     6841 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     6901 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     6961 agagtgcacc atatggacat attgtcgtta gaacgcggct acaattaata cataacctta
     7021 tgtatcatac acatacgatt taggtgacac tata
//
