LOCUS       sequence-369793-        2916 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Markerless yeast integration Spacer at Int.10 locus to used in the 
            Ellis MYT system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2916)
  AUTHORS   Shaw WM, Khalil AS, Ellis T
  TITLE     A Multiplex MoClo Toolkit for Extensive and Flexible Engineering of 
            Saccharomyces cerevisiae.
  JOURNAL   ACS Synth Biol. 2023 Nov 6. doi: 10.1021/acssynbio.3c00423.
  PUBMED    37930278
REFERENCE   2  (bases 1 to 2916)
  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 Nov 6. doi: 10.1021/acssynbio.3c00423."
COMMENT     Sequence Label: pMYT094
FEATURES             Location/Qualifiers
     source          1..2916
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..591)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(83..102)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(805..824)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             816..1631
                     /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             816..1631
                     /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(889..908)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
       61 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      121 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      181 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      241 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      301 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      361 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      421 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      481 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      541 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      601 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      661 gtcatgactg atccttcaac tcagcaaaag ttcgatttat tcaacaaagc cacgttgtgt
      721 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac
      781 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt
      841 cttgctccag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
      901 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg
      961 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     1021 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     1081 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg
     1141 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     1201 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgcc
     1261 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     1321 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataagctt ttgccattct
     1381 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     1441 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     1501 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     1561 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     1621 agtttttcta atcagaattg gttaattggt tgtaacacgc ggccgcctct ttcctctaca
     1681 aaagtatgtt aaatgtaggt agttcaaatt gcggtcacgt atatgccaac gttgttaata
     1741 tataatgatc cattggaaca atgaagttta cagaagctca aaattaatgc aagaataatt
     1801 acgtcaaggg agtcataaca aactaagaaa acaaaggaat ttgactaagt ttgagcgcaa
     1861 tgatatatcc atgcttgata ttctgattca tacatatcgt ttctgtcatt tcaatggaag
     1921 tctttattta tcaattacca caacctttag tattgtgctt attattggtc aaaaggagtt
     1981 catgcgctag tgatagtcaa cccatgaagt gattagtgga acatagaagg ggataaattt
     2041 cctatcggat aacaaagaaa aaccaggatt attgttttac caccacattt tcaaactttc
     2101 ccgctgaatt tcctactacc ctcatttgaa agaacgcttt ccactcgggg taattctagc
     2161 ggtcatactc atgtcttcgt cagaagacat tgtactgact gcagcagtac tagtccgcac
     2221 gatcaagtgt aatcagctct agactgagaa ttccgtagcg taggtgagga agatttaggt
     2281 tggcgttgta tcatcttttc ttccattcaa tgcaccaaga cttaattttg ttggttaaac
     2341 tcttattctt tgtcagttta accaacaaaa ttacctacgg taattagtga aaggccaaaa
     2401 tctaatgtta caatatgatt caatcattct tattgcggag aagctaccga gaagaaccct
     2461 accactggta caacacaagg gaacggtttc attgacctcg tgcataaccg ggaaggtaac
     2521 acatgaatat gctgtgtttg aatcatctcc aatccaaact ataggaatga aaaatttttt
     2581 ttgggtacaa tgttttctca caatatcgta ttcttggtca gcaatgtttc caaggatgac
     2641 agactctttc taagaaaaga tttcaagttc aaaatttaca taacagtgaa ctatgcagaa
     2701 aattttactt ctggaccacg catttaacaa taatgtttgc ccgcaacaat aaagtatttg
     2761 gtcgctttaa gacgttatca gttattttat gagaaggtgt ttaccattcg tgcggccgca
     2821 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2881 aaggccagga accgtaaaaa ggccgcgttg ctggcg
//
