LOCUS       sequence-369786-        2933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Markerless yeast integration Spacer at Int.5 locus to used in the 
            Ellis MYT system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2933)
  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 2933)
  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: pMYT089
FEATURES             Location/Qualifiers
     source          1..2933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(56..644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(136..155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(858..877)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             869..1684
                     /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             869..1684
                     /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(942..961)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
       61 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      121 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
      181 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
      241 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
      301 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
      361 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
      421 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
      481 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
      541 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
      601 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
      661 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
      721 ctgatccttc aactcagcaa aagttcgatt tattcaacaa agccacgttg tgtctcaaaa
      781 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
      841 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc
      901 caggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
      961 taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga
     1021 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     1081 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     1141 tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga
     1201 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     1261 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gcctcgctca
     1321 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     1381 tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat tctcaccgga
     1441 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     1501 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     1561 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     1621 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     1681 ctaatcagaa ttggttaatt ggttgtaaca cgcggccgca acttttgatg attgctggga
     1741 aacggtttgg taaactgctg aagcaattgt tgtacacaat acttctgctt ttggccccat
     1801 ctggattttt catttccgct tttgcctcag gtgcaacaat aactatcttt gagcatcgta
     1861 ccaccaatac aaaaggatat gatcgttatc atacagaatt tttcatggat tttgcatgtt
     1921 tcggtataac tttacgtagg tgaattattg aagggctttg tgataggtgt ggatgtacct
     1981 gccgtgcgtt gatgaccttc aactgatcgg aacaaattgc tattacatgt aaaaatctca
     2041 tgtatgagcg aatctttttg tgttataggc aatcacctag aaaagcttgg aaattggtcc
     2101 tgagtttttt gttgatggta agtgcatttt tttagtatga aaggtttacg atattagctg
     2161 ctaagttctc ccttgcccta gtgaattacg tttttttcga ttaaaagggt tggtcccgaa
     2221 attgacctct tcgcagacag aggaatgaat cactcatgtc ttcctaggaa gacattgtat
     2281 acgctgcagc tgaactagtc cgcctagcag acccagccaa gttctagata gcgaattcag
     2341 atgcccactt ctgataggat atgtacctca attgcagata gcatcaaatt tagactacgt
     2401 catcaaccaa attacaccct atgaaaaaca taagattata agcatgcgtt tgtgtaaagg
     2461 tcataccatt ttatacgttg acgcaatggt gcaaatagaa aggtttacga aagctcttat
     2521 tccgcttagc gcaattctat tgttaatttc aagtaaaaag ataaatatct cacgtcctta
     2581 tattatcttt gattaaaatt tcacgatcaa tgtaacgaaa aaaggtcgca aatatttctt
     2641 tttcatcact ctgattaaac aatggcatca tgaaacgctc attggatcgt tacacgttct
     2701 cattgtacat atgtatgatt tagaacagct tgttcgcctg atctccacct tataccctca
     2761 tcttaccgca gtgcggtttt acgcctcatg ttatttttgc cagctttaat aacaacatca
     2821 gtaatattac gttattggat cttccactcc ggttcgagga aaaaaagaga ggaggagaaa
     2881 cgcggccgca cggttatcca cagaatcagg ggataacgca ggaaagaaca tgt
//
