LOCUS       sequence-369789-        2913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Markerless yeast integration Spacer at Int.7 locus to used in the 
            Ellis MYT system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2913)
  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 2913)
  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: pMYT091
FEATURES             Location/Qualifiers
     source          1..2913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(831..1419)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(911..930)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1633..1652)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1644..2459
                     /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             1644..2459
                     /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(1717..1736)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 accttatagc aaatagcatt ttcaataaaa atgtgacata cagacttcca cttttcttgc
       61 ttttctcatg ctctgtgtga tgctactgtt aaatcctcca cctcttaccc gactcatgtc
      121 ttctagcgaa gacattgtaa tgcctgcagg tcaactagtc cgcggcggag gactttacgg
      181 tgtctagatg acgaattcgg cacggtatgt tttaaggccc gcggcaccac attttggcat
      241 acgtcgggac attggcggct gaatagatta tacccgtcgc ccttttttca tctctccttt
      301 tctttttgtc gtaagcaaat tacagaattg aatatttgaa taaccctaaa aaaaatgcca
      361 ctcccctaaa aacaaaccag attaagttgc aagaaataga ctgattggaa catacgggtc
      421 ctattcagtg ctcaataaaa attaaaataa tgtgatcaaa caacccgaga ctattcttga
      481 attagcatta gccactataa attttaacat ttttcagagt ctttcgtgcg gggaccgtaa
      541 atattagcat tgaccttgtc aatgaagtaa ttgtctttga ttttaaccgt gttaaagatt
      601 tcgcaaaaga gaaagcttgc tggcttggag cagtaacttt ttttgatttt tttttacaag
      661 cacacatgct tttgccttta gaatattacg gctcctcatg tggatagtaa aaacttgacc
      721 gttgcggccg cacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
      781 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
      841 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
      901 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      961 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1021 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1081 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1141 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1201 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1261 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1321 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1381 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1441 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgactgat
     1501 ccttcaactc agcaaaagtt cgatttattc aacaaagcca cgttgtgtct caaaatctct
     1561 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     1621 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtct tgctccaggc
     1681 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     1741 tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt
     1801 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     1861 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     1921 atgcatggtt actcaccact gcgatccccg ggaaaacagc attccaggta ttagaagaat
     1981 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     2041 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgcctc gctcaggcgc
     2101 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     2161 ggcctgttga acaagtctgg aaagaaatgc ataagctttt gccattctca ccggattcag
     2221 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     2281 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     2341 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     2401 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     2461 cagaattggt taattggttg taacacgcgg ccgctacgcc ccagctttac tacgtggatg
     2521 atacagtaag cacatacata ttgacggcat ctaatcgttc taaacggaaa aataggcgct
     2581 gatttgatcc acttcggtta ggtaaccgct gatgccaacg atctagggcc cttccatttt
     2641 aggaaaaagg gtatatactc tgatgccctg tctggccgaa cattcgtata tgtggatgta
     2701 tcgtgatccg ttcgtattca ataaaagtgt aacgatctca tctcaagtca aaagtgagaa
     2761 gggaataact aaaaaagaaa aactgatatt ttgacgtgac gtggtttatt agcaagtatg
     2821 tttcaaccat tctcagcgta aaccctatca caataacgcc tctatgaaaa gtttaacatt
     2881 ttgtctatct cggttcaatc agttgaatag cga
//
