LOCUS       sequence-369784-        3045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Markerless yeast integration Spacer at Int.3 locus to used in the 
            Ellis MYT system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3045)
  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 3045)
  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: pMYT087
FEATURES             Location/Qualifiers
     source          1..3045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     342..361
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1858..2673)
                     /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(1858..2673)
                     /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     2581..2600
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2665..2684
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(2898..3045,1..441)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
       61 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
      121 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
      181 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
      241 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
      301 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
      361 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
      421 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
      481 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
      541 gcggccgcat attttaaatg tttgtatctt gttttctctt tcaaaaaaaa aatggtgtca
      601 ttcattattt tatggtcaac cctatacatc aatttttctc tgaaaatatt gacaaataaa
      661 gtagttgatt cttgttctac caattagtga tattatgcat gactgttaac aactttttga
      721 ctaatctctg aaatcatatg aagatcctgc tgcatttcat gcatctaaga aatcaaccta
      781 tatcaacaga tttcaataat tactctaaac ttatgctgta acttagaaag taaccagcct
      841 gtgttgactg attgagttgc gtattaactg cgcctagtca tttcaacact tataatttgc
      901 ttcagcttaa gtgtggttca tctttttttt tctggaaact ttgcatgccc tcaaagcatg
      961 agtagttagt tatctttttg acaatgatct cttttgaaaa tatctactgt agatttgcat
     1021 ggacgcacgt cgcccatacg ccaaactttg gcaatgatac tcgttattcg taatatcagt
     1081 ccgtcaaggt gctgtgattt ctctatttta tattgcctat tattttttca aatgatttga
     1141 gccgttttaa attgagtatg caatgagtct tttgaatcaa cctgctcaga actcacgact
     1201 ccgaattctc tctctagaag gtcaagtaca ttacgctacg gactagttac gctgcagatg
     1261 ctacaatgtc ttccagtgaa gacatgagta gcatcttgtt accgaccaag taaggcagtt
     1321 ccataaccac tgccacgaat acgtttcact accttgaaga atctctaatg taggccgtat
     1381 tcttcgcact tagttctgac gatgtagaca tctcattata taagagcata agcgcctgtt
     1441 tctggaatca tttcttcgtg acccagcttt ttgagttatt tcgcggtatt ttgaaacatt
     1501 tctcgggctt gacgtgaaca tccttatatt tcatgacaaa ctcgatcatt ggaacatccc
     1561 tgcctcgatt ttagagctag tatcaaattt caatctcttt gtgatggagc cccgctccta
     1621 tttcaaaaga gaagtttctt gtatgcatat gttattgaag tctgattata gcaagtgcaa
     1681 tgtcgtttca attattttaa ctatttttag ccatacatgt tagttatcct caaagagagc
     1741 ctccagactg ggaagcagtg tttgtcattt caaataagta gatttcacag tttgtatgat
     1801 tttcgaagcc aggattcatt gggcggccgc gtgttacaac caattaacca attctgatta
     1861 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
     1921 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
     1981 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
     2041 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
     2101 atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa caggccagcc
     2161 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
     2221 ctgagcgagg cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
     2281 caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc
     2341 ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc
     2401 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
     2461 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     2521 ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt
     2581 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggcct
     2641 ggagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     2701 agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag
     2761 attttgagac acaacgtggc tttgttgaat aaatcgaact tttgctgagt tgaaggatca
     2821 gtcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2881 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2941 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3001 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtt
//
