LOCUS       sequence-369781-        2952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Markerless yeast integration Spacer at Int.1 locus to used in the 
            Ellis MYT system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2952)
  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 2952)
  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: pMYT085
FEATURES             Location/Qualifiers
     source          1..2952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(579..1167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(659..678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1381..1400)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1392..2207
                     /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             1392..2207
                     /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(1465..1484)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gcttatagag gagtttaacc atgaaattaa atgattctcc agcaagaaaa aaaaaactcc
       61 tgactcagtt catgtggcgg gaaatttctc tcattgtctc cccctgcggc gctactaatc
      121 ttaaaggtag tggtggcttc gacattatat cgttggttca ggttgtccga tcactaatga
      181 tctgttactt tattgctatg ttatgtgtca aaaggtgata tccccatttt tactgatcgc
      241 agtaatattt taacctgcca tggttaaatt gtttattgtc attgagttga gcactttttc
      301 attgcactgt agttgttgcg tatttcagac ttactcttat agtgacgcta acaggaggtc
      361 tggctcttaa tacaaggacg tcgtaatatt ctgttcaccg gactcgaaag caagaaggaa
      421 aaaagcacat tgtagccttt tcaaaaacct atcttttggt actaatttat ggcggccgca
      481 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
      541 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
      601 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
      661 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
      721 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
      781 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
      841 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      901 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
      961 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     1021 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1081 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1141 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1201 gctcagtgga acgaaaactc acgttaaggg attttggtca tgactgatcc ttcaactcag
     1261 caaaagttcg atttattcaa caaagccacg ttgtgtctca aaatctctga tgttacattg
     1321 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     1381 caaggggtgt tatgagccat attcaacggg aaacgtcttg ctccaggccg cgattaaatt
     1441 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     1501 gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     1561 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     1621 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     1681 tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat cctgattcag
     1741 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     1801 gtaattgtcc ttttaacagc gatcgcgtat ttcgcctcgc tcaggcgcaa tcacgaatga
     1861 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     1921 aagtctggaa agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg
     1981 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     2041 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     2101 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     2161 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta
     2221 attggttgta acacgcggcc gcaacagtgg ggaaatagtt tacgagtttt gttcaagtta
     2281 aaagcatttt ttgttgtgaa gagaataaac ttgatttcga aggagatcaa attcgaaata
     2341 atctgcgaat ttattatgat cgactaggca acaaagacgc agttttctat taaatctgtg
     2401 cgtttgagta acttaactga ctataatatc gcaatcttaa catcattaaa caatgtaaaa
     2461 gaggtcatac gctataagaa gttgatttga tcccccgtgg attttgaaaa atgtcaagat
     2521 cagcatattg acccatgttt ttccgctcaa tccagtctat caatcatttt tcaacaagga
     2581 taaccataac attcacccac catgctttgg ttcataagat atacggggag atcagcaact
     2641 cttcaactta ttgaacatgt tgagagcagg attaaataat caatgtcctg tgattgcaac
     2701 atctatcatg cctgacgttt ctcgaacgat acaatctaca atttaatgag ccattttagc
     2761 atattaacga gggctgccgt ttccttcgtg cttagacact catgtcttcc agtgaagaca
     2821 ttgtaacgtc tgcaggtaca ctagtccgac gggttaacat attgagactc tagattggga
     2881 attcagtcgt ggatgtgatg ggtaatgagg atcgaatgga caaaagtgtt ttcaggcatg
     2941 ggagaatggg at
//
