LOCUS       sequence-392533-        5632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Markerless yeast integration Vector at Int.8 locus to be 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 5632)
  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 5632)
  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: pMYT082
FEATURES             Location/Qualifiers
     source          1..5632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             545..556
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     RBS             929..940
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1654..1725
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1741..1768
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3905..4720)
                     /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(3905..4720)
                     /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     4628..4647
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4712..4731
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4945..5533
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5434..5453
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcggccgcta aataaaagta attctggtct tgaccaaccg tgatgttcaa aatcactcga
       61 ccaattttca ctacgtcgtc ctgacagccc atagatggga taccagttca ttagtccaag
      121 gagtagcttc cttcaagatt ttctttttta agtagtaaat ccatatcaag aagtgttatg
      181 aaacatatta ttggaggcat gcactattac ctaacgtcgc catatgcgct cgtatcgatt
      241 gtatcgattg ttacgggttc tcttccaccc ctgtagcgct gtctgtgaag ctttcttatt
      301 acacatctcc gcgttcattg acgtcaatct ttttagaagg ggtcctgatc acgaaggaac
      361 gcatagtgca tcgcttaacc tggctagttg gttgcaaagg aatgctattt ttaaatacta
      421 gcgtcttgca aataaacaat cggatatgaa aacaatatga cgtattatat cggctcgtat
      481 taattacatt atactgattt ttaaaggaat catatagcat gggtctttgc acaatgcatt
      541 aatgattgaa ggcagaggca cgaattctac gtctagatag cctgatgaga cgtcagaacg
      601 tgagaccctg tggtcaaaag tgcgttggct ggtagagcca caaacagccg gtacaagcaa
      661 cgatctccag gaccatctga atcatgcgcg gatgacacga actcacgacg gcgatcacag
      721 acattaaccc acagtacaga cactgcgaca acgtggcaat tcgtcgcaat acgcggccgc
      781 gaaagtgaaa cgtgatttca tgcgtcattt tgaacatttt gtaaatctta tttaataatg
      841 tgtgcggcaa ttcacattta atttatgaat gttttcttaa catcgcggca actcaagaaa
      901 cggcaggttc ggatcttagc tactagagaa agaggagaaa tactagatgg ttagcaaagg
      961 cgaggcggtt atcaaggagt ttatgcgttt taaggttcac atggagggta gcatgaacgg
     1021 tcacgagttc gagatcgagg gtgaaggcga gggtcgtccg tacgaaggca cccagaccgc
     1081 gaagctgaaa gtgaccaagg gtggcccgct gccgttcagc tgggacatcc tgagcccgca
     1141 gttcatgtat ggcagccgtg cgtttaccaa acacccggcg gacattccgg attactataa
     1201 gcaaagcttc ccggaaggtt ttaaatggga gcgtgttatg aacttcgaag atggtggcgc
     1261 ggtgaccgtt acccaggaca ccagcctgga ggatggcacc ctgatttaca aggtgaaact
     1321 gcgtggcacc aactttccgc cggatggtcc ggttatgcag aagaaaacga tgggttggga
     1381 agcgagcacc gagcgtctgt atccggaaga tggcgtgctg aagggtgata tcaaaatggc
     1441 gctgcgtctg aaggacggtg gccgttacct ggcggatttt aagaccacct ataaagcgaa
     1501 gaaaccggtg caaatgccgg gtgcgtacaa cgttgaccgt aaactggata ttaccagcca
     1561 caacgaggat tataccgtgg ttgagcaata tgagcgtagc gagggtcgcc acagcaccgg
     1621 cggcatggac gaactgtata agtgaccagg catcaaataa aacgaaaggc tcagtcgaaa
     1681 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact
     1741 ggctcacctt cgggtgggcc tttctgcgtt tatacattca tctttcacct gccattagta
     1801 acccgacttc tcattgagcg ggttacggca gccacaggcc acattccgaa tgtctgggtg
     1861 agcggtccct tttccagcat ccactaaata tctcggatcg cgctttttaa tctggcttcc
     1921 tgaaaaaaat caatggagtg atgcaaactg actggagcaa aaagctgaca caaggcaatc
     1981 gacctacgtg tctgtctatt ttctcacacc ttctattacc ttctaactct ctgggttgga
     2041 aaaaactgaa aaaaaggttg tctccagttt ccacaaatca tccccctgtt tgattaataa
     2101 atatataaag acgacaacta tcgatcataa actcataaaa ctataactcc tttacacttc
     2161 ttattttata gttattctat tttaattctt attgatttta aaaccccaag aacttagttt
     2221 cgaaaacaca cacacacaaa caattaaaat ggtgtctaaa ggtgaagccg tcatcaaaga
     2281 atttatgaga ttcaaggttc acatggaagg ttctatgaac ggtcacgaat ttgaaattga
     2341 gggtgaaggt gagggtagac catacgaagg tacacaaact gctaaattga aggttacaaa
     2401 aggtggtcct ttgccattct cttgggatat tttgtctcca caatttatgt acggttcaag
     2461 agcttttaca aaacacccag ctgatatccc agattattat aagcaatcat ttccagaggg
     2521 tttcaagtgg gaaagagtca tgaattttga agatggtggt gccgttactg ttactcaaga
     2581 tacttcattg gaagatggca ccttgatcta caaggttaag ttaagaggta ctaatttccc
     2641 accagatggt cctgttatgc aaaaaaagac tatgggatgg gaagcttcta ctgaaagatt
     2701 atatccagag gatggtgttt tgaaaggtga tattaagatg gccttgagat tgaaagacgg
     2761 tggtagatac ttggctgatt ttaagactac ttacaaggcc aaaaagccag ttcaaatgcc
     2821 tggtgcttat aacgttgata gaaaattgga tatcacctct cacaacgaag attacaccgt
     2881 tgttgaacag tacgaaagat cagaaggtag acattctact ggtggtatgg atgagttgta
     2941 caagtagctg gttgatggaa aatataattt tattgggcaa acttttgttt atctgatgtg
     3001 ttttatacta ttatcttttt aattaatgat tctatataca aacctgtata ttttttcttt
     3061 aaccaatttt tttttttata gacctagagc tgtactgcgg ccgcgaaatc tgctcgtcag
     3121 tggtgctcac actgacgaat catgtacaga tcataccgat gactgcctgg cgactcacaa
     3181 ctaagcaaga cagccggaac cagcgccggc gaacaccact gcatatatgg catatcacaa
     3241 cagtccagcg ggtatttggt gctcgtctgg gtctcagctg tcgacgtctc aagcatgaca
     3301 ctagttgcac tgcaggtact acaatgtctt cgtcagaaga catgagtcgc taggaaagat
     3361 acacggccgt gggagcacat ccctttgaaa cgcacccacg catttgctaa agaaaaagaa
     3421 agtaataaaa taaaagaggg caaaagaaag aaggacaaga gaaattcttc gccacggcat
     3481 tgaatctatt tctccttacg aaaaaaaaga tcaaggaaac atgtcggaaa caaatgggaa
     3541 acaacaaaaa ggacttatca ttcctacttt tttgtcgagg gcatttgtcg agggtaatac
     3601 tgcatactgc ctattgtcgg tcccgcagaa ttaatagtgg ctgctcatac acaacgaaaa
     3661 aaattgttaa caaaatagaa ggattcgtct atgccctttt ttcagtgcgg aaattatata
     3721 ccacacaagc cgtcgaagcc catagggtcc tttaacaata aagactctcc taaaaaaatt
     3781 tttcttgtga cacgtcagtg caggatataa atgtagcaag cgtatataac aaatgacaaa
     3841 gggttcttgc gcaaggttac tttagcgttg cggccgcgtg ttacaaccaa ttaaccaatt
     3901 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     3961 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     4021 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     4081 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     4141 cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag
     4201 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     4261 attgcgcctg agcgaggcga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     4321 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     4381 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     4441 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     4501 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     4561 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     4621 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     4681 gcggcctgga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     4741 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     4801 atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt gctgagttga
     4861 aggatcagtc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4921 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4981 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5041 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5101 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5161 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5221 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5281 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5341 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5401 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5461 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5521 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5581 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gt
//
