LOCUS       sequence-369782-        2924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Markerless yeast integration Spacer at Int.2 locus to used in the 
            Ellis MYT system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2924)
  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 2924)
  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: pMYT086
FEATURES             Location/Qualifiers
     source          1..2924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(456..1044)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(536..555)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1258..1277)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1269..2084
                     /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             1269..2084
                     /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(1342..1361)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gatttacatt gaggggggaa acttgcttta tccgctcctc caattgattc aagaagtatg
       61 ggcgctgacc taccatctca atttcttgtt tcactcggga attgtcactg agatagtaaa
      121 tgtctggaga atcctttgca acaggagatt tcaaatttat attctttgtg ttcgcattcg
      181 cttcccgtct gttactcgtt cccttaccat tctattatgc attgcaggtc tgccaagtaa
      241 aaaaaaaaat accttggtgt acacaccacc catacattat ctgacaagga aacaaaagta
      301 atgacagtaa attgaagaac ttgtgatgag ccaaaaagaa gaatgtgtgc ggccgcacgg
      361 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
      421 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
      481 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
      541 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
      601 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
      661 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
      721 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
      781 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
      841 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
      901 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
      961 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     1021 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     1081 cagtggaacg aaaactcacg ttaagggatt ttggtcatga ctgatccttc aactcagcaa
     1141 aagttcgatt tattcaacaa agccacgttg tgtctcaaaa tctctgatgt tacattgcac
     1201 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     1261 ggggtgttat gagccatatt caacgggaaa cgtcttgctc caggccgcga ttaaattcca
     1321 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     1381 cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     1441 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     1501 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     1561 ccactgcgat ccccgggaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     1621 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     1681 attgtccttt taacagcgat cgcgtatttc gcctcgctca ggcgcaatca cgaatgaata
     1741 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     1801 tctggaaaga aatgcataag cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     1861 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     1921 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     1981 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     2041 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     2101 ggttgtaaca cgcggccgct ttcgtcacat gttcaactaa attccaaaca gttgaacttt
     2161 ttgttctact gatcttgtaa ttcatgcttc cttctatgca acacgccata ctaatttagt
     2221 agaatgaaca gtaactggtg aacgatgact ctgtagtaga atactataaa aaggtggcat
     2281 cgtataaagg tgccttcttt gacccttggc aggtagggcg gcaacggcac aatgatattt
     2341 tatttaacaa aaagatacgc taacaccgca tctgaacacg taatactact atatgacttg
     2401 gaacaaatgg ctaatcaata atcacatatg ttcgaccgcc gcaggaaaag agaaaagaaa
     2461 gcagtttcta caagcaagaa cttggaaata cagacaaacc tccagtactt tattaggatt
     2521 gtccattcct cacccagata tgttaaattc gataccaatg tgttttgatt cacgcgcaga
     2581 aagaaggact ttccctgctc tgctcggcaa aaacatacac gttgtcgacc gagcacgact
     2641 gccgttgtcg aagccaccaa gtccaaagta acggcatgga ctcatgtctt cgactgaaga
     2701 cattgtaagc tctgcagtga cactagtccg cggtgtccct gcgctcatct tctagaagtc
     2761 gaattcggga taggggagac aagaacctgt cacttctcgt ttaatatctt taatcgcatc
     2821 atctatagtt cctaattctg cattcctaac actgtagcct atccagcaaa atcattccca
     2881 ttttctcttt atatgtgcct agattttaaa attttgctta agat
//
