LOCUS       sequence-369759-        5620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Markerless yeast integration Vector at Int.4 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 5620)
  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 5620)
  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: pMYT078
FEATURES             Location/Qualifiers
     source          1..5620
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1539..1566)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(1582..1653)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             2367..2378
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      complement(3382..3970)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3462..3481)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4184..4203)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4195..5010
                     /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             4195..5010
                     /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(4268..4287)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 aactagtcgt atgcttgaga cgtgaccagc tgagacccag acgagcacca aatacccgct
       61 ggactgttgt gatatgccat atatgcagtg gtgttcgccg gcgctggttc cggctgtctt
      121 gcttagttgt gagtcgccag gcagtcatcg gtatgatctg tacatgattc gtcagtgtga
      181 gcaccactga cgagcagatt tcgcggccgc agtacagctc taggtctata aaaaaaaaaa
      241 ttggttaaag aaaaaatata caggtttgta tatagaatca ttaattaaaa agataatagt
      301 ataaaacaca tcagataaac aaaagtttgc ccaataaaat tatattttcc atcaaccagc
      361 tacttgtaca actcatccat accaccagta gaatgtctac cttctgatct ttcgtactgt
      421 tcaacaacgg tgtaatcttc gttgtgagag gtgatatcca attttctatc aacgttataa
      481 gcaccaggca tttgaactgg ctttttggcc ttgtaagtag tcttaaaatc agccaagtat
      541 ctaccaccgt ctttcaatct caaggccatc ttaatatcac ctttcaaaac accatcctct
      601 ggatataatc tttcagtaga agcttcccat cccatagtct ttttttgcat aacaggacca
      661 tctggtggga aattagtacc tcttaactta accttgtaga tcaaggtgcc atcttccaat
      721 gaagtatctt gagtaacagt aacggcacca ccatcttcaa aattcatgac tctttcccac
      781 ttgaaaccct ctggaaatga ttgcttataa taatctggga tatcagctgg gtgttttgta
      841 aaagctcttg aaccgtacat aaattgtgga gacaaaatat cccaagagaa tggcaaagga
      901 ccaccttttg taaccttcaa tttagcagtt tgtgtacctt cgtatggtct accctcacct
      961 tcaccctcaa tttcaaattc gtgaccgttc atagaacctt ccatgtgaac cttgaatctc
     1021 ataaattctt tgatgacggc ttcaccttta gacaccattt taattgtttg tgtgtgtgtg
     1081 ttttcgaaac taagttcttg gggttttaaa atcaataaga attaaaatag aataactata
     1141 aaataagaag tgtaaaggag ttatagtttt atgagtttat gatcgatagt tgtcgtcttt
     1201 atatatttat taatcaaaca gggggatgat ttgtggaaac tggagacaac ctttttttca
     1261 gttttttcca acccagagag ttagaaggta atagaaggtg tgagaaaata gacagacacg
     1321 taggtcgatt gccttgtgtc agctttttgc tccagtcagt ttgcatcact ccattgattt
     1381 ttttcaggaa gccagattaa aaagcgcgat ccgagatatt tagtggatgc tggaaaaggg
     1441 accgctcacc cagacattcg gaatgtggcc tgtggctgcc gtaacccgct caatgagaag
     1501 tcgggttact aatggcaggt gaaagatgaa tgtataaacg cagaaaggcc cacccgaagg
     1561 tgagccagtg tgactctagt agagagcgtt caccgacaaa caacagataa aacgaaaggc
     1621 ccagtctttc gactgagcct ttcgttttat ttgatgcctg gtcacttata cagttcgtcc
     1681 atgccgccgg tgctgtggcg accctcgcta cgctcatatt gctcaaccac ggtataatcc
     1741 tcgttgtggc tggtaatatc cagtttacgg tcaacgttgt acgcacccgg catttgcacc
     1801 ggtttcttcg ctttataggt ggtcttaaaa tccgccaggt aacggccacc gtccttcaga
     1861 cgcagcgcca ttttgatatc acccttcagc acgccatctt ccggatacag acgctcggtg
     1921 ctcgcttccc aacccatcgt tttcttctgc ataaccggac catccggcgg aaagttggtg
     1981 ccacgcagtt tcaccttgta aatcagggtg ccatcctcca ggctggtgtc ctgggtaacg
     2041 gtcaccgcgc caccatcttc gaagttcata acacgctccc atttaaaacc ttccgggaag
     2101 ctttgcttat agtaatccgg aatgtccgcc gggtgtttgg taaacgcacg gctgccatac
     2161 atgaactgcg ggctcaggat gtcccagctg aacggcagcg ggccaccctt ggtcactttc
     2221 agcttcgcgg tctgggtgcc ttcgtacgga cgaccctcgc cttcaccctc gatctcgaac
     2281 tcgtgaccgt tcatgctacc ctccatgtga accttaaaac gcataaactc cttgataacc
     2341 gcctcgcctt tgctaaccat ctagtatttc tcctctttct ctagtagcta agatccgaac
     2401 ctgccgtttc ttgagttgcc gcgatgttaa gaaaacattc ataaattaaa tgtgaattgc
     2461 cgcacacatt attaaataag atttacaaaa tgttcaaaat gacgcatgaa atcacgtttc
     2521 actttcgcgg ccgcgtattg cgacgaattg ccacgttgtc gcagtgtctg tactgtgggt
     2581 taatgtctgt gatcgccgtc gtgagttcgt gtcatccgcg catgattcag atggtcctgg
     2641 agatcgttgc ttgtaccggc tgtttgtggc tctaccagcc aacgcacttt tgaccacagg
     2701 gtctcacgtt gtaccgtctc atcagtagct ctagaatgcg aattctgtca ctctcactcc
     2761 aagcccagag gaacaatcta tttacagtag cgctgatggc atcttgtgtg aaactaatgg
     2821 tatttgcgta gacaaaaccc aatacacatc taaattgtcg atacttgtga ggtgtggata
     2881 ttctaacgat caaaatagct tagatgaaac gacttgctgt gctgcgggtg tgactgaatg
     2941 tattttcaac gtactgaatg tactctcaac gtgggcgtga agttgagggc tgggcagaat
     3001 ctggagtgga tcatagcaaa tatgttaaac atagaagtaa actgagagac tgacgagttt
     3061 cttttgtcaa caggcgttga atcttgttag atattaacga ggtggatttt tggcaggtaa
     3121 tttcgcaacc cgacttttca atttgcgctt cccatatttt acactttttt tagtcagtaa
     3181 atgcaaacga cagcaaataa atgaattaaa tcagctaaga aaggaaaaaa attagaagga
     3241 aatggtagtt aaaacatgga caatttagtg tattgcggcc gcacggttat ccacagaatc
     3301 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3361 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3421 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3481 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3541 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3601 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3661 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3721 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3781 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3841 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3901 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3961 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4021 ctcacgttaa gggattttgg tcatgactga tccttcaact cagcaaaagt tcgatttatt
     4081 caacaaagcc acgttgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     4141 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     4201 catattcaac gggaaacgtc ttgctccagg ccgcgattaa attccaacat ggatgctgat
     4261 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga
     4321 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     4381 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     4441 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     4501 gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     4561 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     4621 agcgatcgcg tatttcgcct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     4681 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     4741 cataagcttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     4801 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     4861 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     4921 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     4981 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacacgcg
     5041 gccgcatcaa gttatccgtg gcatactttg aatggaaact gaattaactg ctgtgataca
     5101 gaacaaaaat aaagaggatg gcagggagag gaaatgataa aataagttac gccacttaag
     5161 caaaaatgat agatgaaggg cccaattaaa gatcactttt ttcttagcga agtatgcggg
     5221 agaatgcttg attgactctc gtaaaggctc tcgaaaaaaa taggcgcgtt tagtaacaaa
     5281 gaacatggta gtacaattga ttaatacttt ttttttctgt tttagcagtg gagctcttag
     5341 cgaaggagcg atgtgctgtg ggacttctca acccccttca ttctgtaatt tgcctaattc
     5401 atattatgag gatttgtcgt cctaaaattt ggaagtagca tatttcttct tcactcaaga
     5461 agaaataccc ctccagatat tagctttatt ctggtggcct gtcactgtca catatattat
     5521 tcaatgatta ttggttttct agttccattc gcacgataaa ctaactcagg tccgtgtcga
     5581 ctcatgtctt catcggaaga cattgtacat gctgcaggct
//
