LOCUS       sequence-369798-        5610 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Markerless yeast integration Vector at Int.9 locus to used in the 
            Dueber YTK and Ellis MYT systems.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5610)
  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 5610)
  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: pMYT083
FEATURES             Location/Qualifiers
     source          1..5610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             896..907
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1621..1692
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1708..1735
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3883..4698)
                     /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(3883..4698)
                     /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     4606..4625
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4690..4709
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4923..5511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5412..5431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcggccgcca gatgatcacg aatcccgaag cgatgtaatt ttccgtgagg cagtcctgaa
       61 tcaggccgta gattgactat tagcatctct ttttgagtat gctaagaagt agagggctac
      121 cttattccca cacacttttt accacgggcg aaacatttat tttcaagtcc cagcacagtg
      181 atgggatgaa gtccaggatg cgtcatacgg tcctatggca aacttcgggg aaggtttaat
      241 atgctgcttt caagcaggat taacggtaat caactcccta ggcaaattgg ctcaactctg
      301 cctatctatc cactgaggat ttcaaatcca ccgaatggga gaatatgtca gtgtccattt
      361 tggagtaagg tatgagttct tttttcgtca gcttttcttc tatatcttac aattgcttat
      421 cttaaaccac cgatattttt gcagtccctt catcatcgag cacaatggcc tatacgttga
      481 aattttggtt ttggtttgaa gcccactccg attcccttgt ccaacaaaga attctagctc
      541 tagattggct gatgagacgt cgaaacgtga gaccctgtgg tcaaaagtgc gttggctggt
      601 agagccacaa acagccggta caagcaacga tctccaggac catctgaatc atgcgcggat
      661 gacacgaact cacgacggcg atcacagaca ttaacccaca gtacagacac tgcgacaacg
      721 tggcaattcg tcgcaatacg cggccgcgaa agtgaaacgt gatttcatgc gtcattttga
      781 acattttgta aatcttattt aataatgtgt gcggcaattc acatttaatt tatgaatgtt
      841 ttcttaacat cgcggcaact caagaaacgg caggttcgga tcttagctac tagagaaaga
      901 ggagaaatac tagatggtta gcaaaggcga ggcggttatc aaggagttta tgcgttttaa
      961 ggttcacatg gagggtagca tgaacggtca cgagttcgag atcgagggtg aaggcgaggg
     1021 tcgtccgtac gaaggcaccc agaccgcgaa gctgaaagtg accaagggtg gcccgctgcc
     1081 gttcagctgg gacatcctga gcccgcagtt catgtatggc agccgtgcgt ttaccaaaca
     1141 cccggcggac attccggatt actataagca aagcttcccg gaaggtttta aatgggagcg
     1201 tgttatgaac ttcgaagatg gtggcgcggt gaccgttacc caggacacca gcctggagga
     1261 tggcaccctg atttacaagg tgaaactgcg tggcaccaac tttccgccgg atggtccggt
     1321 tatgcagaag aaaacgatgg gttgggaagc gagcaccgag cgtctgtatc cggaagatgg
     1381 cgtgctgaag ggtgatatca aaatggcgct gcgtctgaag gacggtggcc gttacctggc
     1441 ggattttaag accacctata aagcgaagaa accggtgcaa atgccgggtg cgtacaacgt
     1501 tgaccgtaaa ctggatatta ccagccacaa cgaggattat accgtggttg agcaatatga
     1561 gcgtagcgag ggtcgccaca gcaccggcgg catggacgaa ctgtataagt gaccaggcat
     1621 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     1681 gtgaacgctc tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat
     1741 acattcatct ttcacctgcc attagtaacc cgacttctca ttgagcgggt tacggcagcc
     1801 acaggccaca ttccgaatgt ctgggtgagc ggtccctttt ccagcatcca ctaaatatct
     1861 cggatcgcgc tttttaatct ggcttcctga aaaaaatcaa tggagtgatg caaactgact
     1921 ggagcaaaaa gctgacacaa ggcaatcgac ctacgtgtct gtctattttc tcacaccttc
     1981 tattaccttc taactctctg ggttggaaaa aactgaaaaa aaggttgtct ccagtttcca
     2041 caaatcatcc ccctgtttga ttaataaata tataaagacg acaactatcg atcataaact
     2101 cataaaacta taactccttt acacttctta ttttatagtt attctatttt aattcttatt
     2161 gattttaaaa ccccaagaac ttagtttcga aaacacacac acacaaacaa ttaaaatggt
     2221 gtctaaaggt gaagccgtca tcaaagaatt tatgagattc aaggttcaca tggaaggttc
     2281 tatgaacggt cacgaatttg aaattgaggg tgaaggtgag ggtagaccat acgaaggtac
     2341 acaaactgct aaattgaagg ttacaaaagg tggtcctttg ccattctctt gggatatttt
     2401 gtctccacaa tttatgtacg gttcaagagc ttttacaaaa cacccagctg atatcccaga
     2461 ttattataag caatcatttc cagagggttt caagtgggaa agagtcatga attttgaaga
     2521 tggtggtgcc gttactgtta ctcaagatac ttcattggaa gatggcacct tgatctacaa
     2581 ggttaagtta agaggtacta atttcccacc agatggtcct gttatgcaaa aaaagactat
     2641 gggatgggaa gcttctactg aaagattata tccagaggat ggtgttttga aaggtgatat
     2701 taagatggcc ttgagattga aagacggtgg tagatacttg gctgatttta agactactta
     2761 caaggccaaa aagccagttc aaatgcctgg tgcttataac gttgatagaa aattggatat
     2821 cacctctcac aacgaagatt acaccgttgt tgaacagtac gaaagatcag aaggtagaca
     2881 ttctactggt ggtatggatg agttgtacaa gtagctggtt gatggaaaat ataattttat
     2941 tgggcaaact tttgtttatc tgatgtgttt tatactatta tctttttaat taatgattct
     3001 atatacaaac ctgtatattt tttctttaac caattttttt ttttatagac ctagagctgt
     3061 actgcggccg cgaaatctgc tcgtcagtgg tgctcacact gacgaatcat gtacagatca
     3121 taccgatgac tgcctggcga ctcacaacta agcaagacag ccggaaccag cgccggcgaa
     3181 caccactgca tatatggcat atcacaacag tccagcgggt atttggtgct cgtctgggtc
     3241 tcagctgtga ccgtctcaag catgcaacta gtacgtctgc agtacgtaca atgtcttcta
     3301 gcgaagacat gagtacccga aggaaatgtg tagaatcttg gcatcgctga cgtgttttct
     3361 tccaagattt aaagaggact actcggtgca ggtggacatc tcgaatgaat atgttaccta
     3421 gcttgatctc ctccatcaaa ttacaacctt tttgcagtga attttgatac atggatttcg
     3481 gagcttaaac acgtattaac tgaaattcag gcatggcgag aaaattccca tcagaacttg
     3541 aagaatactc taaatactac tgagataaac cgagaggtga cgtgaaattc ttcccagatg
     3601 ttcaatatat tatgaaaaac catccagctg aattacctca tttgcagatc aatttcaaag
     3661 gcttcggttc ggggataatg agacattctg tccaaaatgt cccatttatt acagcctcgg
     3721 aattaaggtg cttgatactg ttctcataaa catgtgactg cattggtggt tgatggagta
     3781 atgataccac tgagcggttt caaatgctaa aagtcttttg aagctatcac agttaccctc
     3841 ggatttggcg gccgcgtgtt acaaccaatt aaccaattct gattagaaaa actcatcgag
     3901 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     3961 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     4021 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     4081 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     4141 caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     4201 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgaggcgaaa
     4261 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     4321 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     4381 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     4441 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     4501 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     4561 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     4621 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctggagc aagacgtttc
     4681 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     4741 tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacaac
     4801 gtggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagtcat gaccaaaatc
     4861 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4921 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4981 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5041 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5101 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5161 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5221 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5281 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5341 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5401 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5461 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5521 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5581 gcgttatccc ctgattctgt ggataaccgt
//
