LOCUS       sequence-392534-        2933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Markerless yeast integration Spacer at Int.8 locus to be 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: pMYT092
FEATURES             Location/Qualifiers
     source          1..2933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             545..556
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(1206..2021)
                     /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             complement(1206..2021)
                     /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     1929..1948
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2013..2032
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2246..2834
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2735..2754
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcggccgcta aataaaagta attctggtct tgaccaaccg tgatgttcaa aatcactcga
       61 ccaattttca ctacgtcgtc ctgacagccc atagatggga taccagttca ttagtccaag
      121 gagtagcttc cttcaagatt ttctttttta agtagtaaat ccatatcaag aagtgttatg
      181 aaacatatta ttggaggcat gcactattac ctaacgtcgc catatgcgct cgtatcgatt
      241 gtatcgattg ttacgggttc tcttccaccc ctgtagcgct gtctgtgaag ctttcttatt
      301 acacatctcc gcgttcattg acgtcaatct ttttagaagg ggtcctgatc acgaaggaac
      361 gcatagtgca tcgcttaacc tggctagttg gttgcaaagg aatgctattt ttaaatacta
      421 gcgtcttgca aataaacaat cggatatgaa aacaatatga cgtattatat cggctcgtat
      481 taattacatt atactgattt ttaaaggaat catatagcat gggtctttgc acaatgcatt
      541 aatgattgaa ggcagaggca cgaattctac gtctagatat gaacaacgca aggattgcgg
      601 actagttgca ctgcaggtac tacaatgtct tcgtcagaag acatgagtcg ctaggaaaga
      661 tacacggccg tgggagcaca tccctttgaa acgcacccac gcatttgcta aagaaaaaga
      721 aagtaataaa ataaaagagg gcaaaagaaa gaaggacaag agaaattctt cgccacggca
      781 ttgaatctat ttctccttac gaaaaaaaag atcaaggaaa catgtcggaa acaaatggga
      841 aacaacaaaa aggacttatc attcctactt ttttgtcgag ggcatttgtc gagggtaata
      901 ctgcatactg cctattgtcg gtcccgcaga attaatagtg gctgctcata cacaacgaaa
      961 aaaattgtta acaaaataga aggattcgtc tatgcccttt tttcagtgcg gaaattatat
     1021 accacacaag ccgtcgaagc ccatagggtc ctttaacaat aaagactctc ctaaaaaaat
     1081 ttttcttgtg acacgtcagt gcaggatata aatgtagcaa gcgtatataa caaatgacaa
     1141 agggttcttg cgcaaggtta ctttagcgtt gcggccgcgt gttacaacca attaaccaat
     1201 tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     1261 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     1321 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     1381 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     1441 acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca
     1501 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     1561 gattgcgcct gagcgaggcg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     1621 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     1681 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     1741 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     1801 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     1861 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     1921 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     1981 cgcggcctgg agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     2041 tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa
     2101 catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg
     2161 aaggatcagt catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2221 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2281 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2341 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     2401 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2461 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2521 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2581 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2641 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2701 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2761 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2821 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2881 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgt
//
