LOCUS       sequence-369758-        5744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Markerless yeast integration Vector at Int.3 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 5744)
  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 5744)
  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: pMYT077
FEATURES             Location/Qualifiers
     source          1..5744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1692..1719)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(1735..1806)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             2520..2531
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      complement(3660..4248)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3740..3759)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4462..4481)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4473..5288
                     /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             4473..5288
                     /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(4546..4565)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 cgtcagaact aagtgcgaag aatacggcct acattagaga ttcttcaagg tagtgaaacg
       61 tattcgtggc agtggttatg gaactgcctt acttggtcgg taacaagatg ctactcatgt
      121 cttcactgga agacattgta gcatctgcag cgtaactagt tgactgcttg agacggtacc
      181 agctgagacc cagacgagca ccaaataccc gctggactgt tgtgatatgc catatatgca
      241 gtggtgttcg ccggcgctgg ttccggctgt cttgcttagt tgtgagtcgc caggcagtca
      301 tcggtatgat ctgtacatga ttcgtcagtg tgagcaccac tgacgagcag atttcgcggc
      361 cgcagtacag ctctaggtct ataaaaaaaa aaattggtta aagaaaaaat atacaggttt
      421 gtatatagaa tcattaatta aaaagataat agtataaaac acatcagata aacaaaagtt
      481 tgcccaataa aattatattt tccatcaacc agctacttgt acaactcatc cataccacca
      541 gtagaatgtc taccttctga tctttcgtac tgttcaacaa cggtgtaatc ttcgttgtga
      601 gaggtgatat ccaattttct atcaacgtta taagcaccag gcatttgaac tggctttttg
      661 gccttgtaag tagtcttaaa atcagccaag tatctaccac cgtctttcaa tctcaaggcc
      721 atcttaatat cacctttcaa aacaccatcc tctggatata atctttcagt agaagcttcc
      781 catcccatag tctttttttg cataacagga ccatctggtg ggaaattagt acctcttaac
      841 ttaaccttgt agatcaaggt gccatcttcc aatgaagtat cttgagtaac agtaacggca
      901 ccaccatctt caaaattcat gactctttcc cacttgaaac cctctggaaa tgattgctta
      961 taataatctg ggatatcagc tgggtgtttt gtaaaagctc ttgaaccgta cataaattgt
     1021 ggagacaaaa tatcccaaga gaatggcaaa ggaccacctt ttgtaacctt caatttagca
     1081 gtttgtgtac cttcgtatgg tctaccctca ccttcaccct caatttcaaa ttcgtgaccg
     1141 ttcatagaac cttccatgtg aaccttgaat ctcataaatt ctttgatgac ggcttcacct
     1201 ttagacacca ttttaattgt ttgtgtgtgt gtgttttcga aactaagttc ttggggtttt
     1261 aaaatcaata agaattaaaa tagaataact ataaaataag aagtgtaaag gagttatagt
     1321 tttatgagtt tatgatcgat agttgtcgtc tttatatatt tattaatcaa acagggggat
     1381 gatttgtgga aactggagac aacctttttt tcagtttttt ccaacccaga gagttagaag
     1441 gtaatagaag gtgtgagaaa atagacagac acgtaggtcg attgccttgt gtcagctttt
     1501 tgctccagtc agtttgcatc actccattga tttttttcag gaagccagat taaaaagcgc
     1561 gatccgagat atttagtgga tgctggaaaa gggaccgctc acccagacat tcggaatgtg
     1621 gcctgtggct gccgtaaccc gctcaatgag aagtcgggtt actaatggca ggtgaaagat
     1681 gaatgtataa acgcagaaag gcccacccga aggtgagcca gtgtgactct agtagagagc
     1741 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct ttcgactgag cctttcgttt
     1801 tatttgatgc ctggtcactt atacagttcg tccatgccgc cggtgctgtg gcgaccctcg
     1861 ctacgctcat attgctcaac cacggtataa tcctcgttgt ggctggtaat atccagttta
     1921 cggtcaacgt tgtacgcacc cggcatttgc accggtttct tcgctttata ggtggtctta
     1981 aaatccgcca ggtaacggcc accgtccttc agacgcagcg ccattttgat atcacccttc
     2041 agcacgccat cttccggata cagacgctcg gtgctcgctt cccaacccat cgttttcttc
     2101 tgcataaccg gaccatccgg cggaaagttg gtgccacgca gtttcacctt gtaaatcagg
     2161 gtgccatcct ccaggctggt gtcctgggta acggtcaccg cgccaccatc ttcgaagttc
     2221 ataacacgct cccatttaaa accttccggg aagctttgct tatagtaatc cggaatgtcc
     2281 gccgggtgtt tggtaaacgc acggctgcca tacatgaact gcgggctcag gatgtcccag
     2341 ctgaacggca gcgggccacc cttggtcact ttcagcttcg cggtctgggt gccttcgtac
     2401 ggacgaccct cgccttcacc ctcgatctcg aactcgtgac cgttcatgct accctccatg
     2461 tgaaccttaa aacgcataaa ctccttgata accgcctcgc ctttgctaac catctagtat
     2521 ttctcctctt tctctagtag ctaagatccg aacctgccgt ttcttgagtt gccgcgatgt
     2581 taagaaaaca ttcataaatt aaatgtgaat tgccgcacac attattaaat aagatttaca
     2641 aaatgttcaa aatgacgcat gaaatcacgt ttcactttcg cggccgcgta ttgcgacgaa
     2701 ttgccacgtt gtcgcagtgt ctgtactgtg ggttaatgtc tgtgatcgcc gtcgtgagtt
     2761 cgtgtcatcc gcgcatgatt cagatggtcc tggagatcgt tgcttgtacc ggctgtttgt
     2821 ggctctacca gccaacgcac ttttgaccac agggtctcac gtttagccgt ctcatcagat
     2881 gctctagaga gagaattcgg agtcgtgagt tctgagcagg ttgattcaaa agactcattg
     2941 catactcaat ttaaaacggc tcaaatcatt tgaaaaaata ataggcaata taaaatagag
     3001 aaatcacagc accttgacgg actgatatta cgaataacga gtatcattgc caaagtttgg
     3061 cgtatgggcg acgtgcgtcc atgcaaatct acagtagata ttttcaaaag agatcattgt
     3121 caaaaagata actaactact catgctttga gggcatgcaa agtttccaga aaaaaaaaga
     3181 tgaaccacac ttaagctgaa gcaaattata agtgttgaaa tgactaggcg cagttaatac
     3241 gcaactcaat cagtcaacac aggctggtta ctttctaagt tacagcataa gtttagagta
     3301 attattgaaa tctgttgata taggttgatt tcttagatgc atgaaatgca gcaggatctt
     3361 catatgattt cagagattag tcaaaaagtt gttaacagtc atgcataata tcactaattg
     3421 gtagaacaag aatcaactac tttatttgtc aatattttca gagaaaaatt gatgtatagg
     3481 gttgaccata aaataatgaa tgacaccatt ttttttttga aagagaaaac aagatacaaa
     3541 catttaaaat atgcggccgc acggttatcc acagaatcag gggataacgc aggaaagaac
     3601 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3661 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3721 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3781 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3841 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3901 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3961 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4021 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4081 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4141 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4201 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4261 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4321 atgactgatc cttcaactca gcaaaagttc gatttattca acaaagccac gttgtgtctc
     4381 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
     4441 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt
     4501 gctccaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     4561 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
     4621 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     4681 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     4741 ctcctgatga tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat
     4801 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     4861 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgcctcg
     4921 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     4981 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac
     5041 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     5101 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     5161 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     5221 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     5281 ttttctaatc agaattggtt aattggttgt aacacgcggc cgcccaatga atcctggctt
     5341 cgaaaatcat acaaactgtg aaatctactt atttgaaatg acaaacactg cttcccagtc
     5401 tggaggctct ctttgaggat aactaacatg tatggctaaa aatagttaaa ataattgaaa
     5461 cgacattgca cttgctataa tcagacttca ataacatatg catacaagaa acttctcttt
     5521 tgaaatagga gcggggctcc atcacaaaga gattgaaatt tgatactagc tctaaaatcg
     5581 aggcagggat gttccaatga tcgagtttgt catgaaatat aaggatgttc acgtcaagcc
     5641 cgagaaatgt ttcaaaatac cgcgaaataa ctcaaaaagc tgggtcacga agaaatgatt
     5701 ccagaaacag gcgcttatgc tcttatataa tgagatgtct acat
//
