LOCUS       sequence-393207-        2593 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Easy-MISE toolkit pEM-plasmid containing mCherry coding sequence 
            with CD protruding ends.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2593)
  AUTHORS   Maestroni L, Butti P, Milanesi R, Pagliari S, Campone L, Serra I, 
            Branduardi P
  TITLE     Easy Modular Integrative fuSion-ready Expression (Easy-MISE) Toolkit
            for Fast Engineering of Heterologous Productions in Saccharomyces 
            cerevisiae.
  JOURNAL   ACS Synth Biol. 2023 Apr 14. doi: 10.1021/acssynbio.3c00015.
  PUBMED    37058502
REFERENCE   2  (bases 1 to 2593)
  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: "ACS Synth 
            Biol. 2023 Apr 14. doi: 10.1021/acssynbio.3c00015."
COMMENT     Sequence Label: pEM.F03L
FEATURES             Location/Qualifiers
     source          1..2593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(139..158)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             150..965
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             150..965
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(223..242)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1050..1638
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1539..1558
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1792..1809
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1877..2584)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2155..2174)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2424..2442
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
ORIGIN
        1 gaccctgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
       61 atttaaaagg atctaggtga agatcctttt tgataatctc atgaacaata aaactgtctg
      121 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct
      181 ctaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
      241 ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag
      301 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
      361 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
      421 ctgatgatgc atggttactc accactgcga tccccgggaa aacagcattc caggtattag
      481 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
      541 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgactcgctc
      601 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
      661 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
      721 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
      781 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
      841 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
      901 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
      961 tctaagaatt aattcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1021 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1081 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1141 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     1201 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     1261 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     1321 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     1381 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     1441 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     1501 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     1561 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     1621 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     1681 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     1741 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     1801 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     1861 attctggtct cccagactac ttgtacagct cgtccatgcc gccggtggag tggcggccct
     1921 cggcgcgttc gtactgttcc acgatggtgt agtcctcgtt gtgggaggtg atgtccaact
     1981 tgatgttgac gttgtaggcg ccgggcagct gcacgggctt cttggccttg taggtggtct
     2041 tgacctcagc gtcgtagtgg ccgccgtcct tcagcttcag cctctgcttg atctcgccct
     2101 tcagggcgcc gtcctcgggg tacatccgct cggaggaggc ctcccagccc atggtcttct
     2161 tctgcattac ggggccgtcg gaggggaagt tggtgccgcg cagcttcacc ttgtagatga
     2221 actcgccgtc ctgcagggag gagtcctggg tcacggtcac cacgccgccg tcctcgaagt
     2281 tcatcacgcg ctcccacttg aagccctcgg ggaaggacag cttcaagtag tcggggatgt
     2341 cggcggggtg cttcacgtag gccttggagc cgtacatgaa ctgaggggac aggatgtccc
     2401 aggcgaaggg cagggggcca cccttggtca ccttcagctt ggcggtctgg gtgccctcgt
     2461 aggggcggcc ctcgccctcg ccctcgatct cgaactcgtg gccgttcacg gagccctcca
     2521 tgtgcacctt gaagcgcatg aactccttga tgatggccat gttatcctcc tcgcccttgc
     2581 tcacagaact aga
//
