LOCUS       sequence-369779-        5615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Markerless yeast integration Vector at Int.10 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 5615)
  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 5615)
  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: pMYT084
FEATURES             Location/Qualifiers
     source          1..5615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             95..106
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      complement(1139..1727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1219..1238)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1941..1960)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1952..2767
                     /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             1952..2767
                     /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(2025..2044)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(4882..4909)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(4925..4996)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tctcgaactc gtgaccgttc atgctaccct ccatgtgaac cttaaaacgc ataaactcct
       61 tgataaccgc ctcgcctttg ctaaccatct agtatttctc ctctttctct agtagctaag
      121 atccgaacct gccgtttctt gagttgccgc gatgttaaga aaacattcat aaattaaatg
      181 tgaattgccg cacacattat taaataagat ttacaaaatg ttcaaaatga cgcatgaaat
      241 cacgtttcac tttcgcggcc gcgtattgcg acgaattgcc acgttgtcgc agtgtctgta
      301 ctgtgggtta atgtctgtga tcgccgtcgt gagttcgtgt catccgcgca tgattcagat
      361 ggtcctggag atcgttgctt gtaccggctg tttgtggctc taccagccaa cgcacttttg
      421 accacagggt ctcacgttgt cacgtctcat cagaacctct agactgagaa ttccgtagcg
      481 taggtgagga agatttaggt tggcgttgta tcatcttttc ttccattcaa tgcaccaaga
      541 cttaattttg ttggttaaac tcttattctt tgtcagttta accaacaaaa ttacctacgg
      601 taattagtga aaggccaaaa tctaatgtta caatatgatt caatcattct tattgcggag
      661 aagctaccga gaagaaccct accactggta caacacaagg gaacggtttc attgacctcg
      721 tgcataaccg ggaaggtaac acatgaatat gctgtgtttg aatcatctcc aatccaaact
      781 ataggaatga aaaatttttt ttgggtacaa tgttttctca caatatcgta ttcttggtca
      841 gcaatgtttc caaggatgac agactctttc taagaaaaga tttcaagttc aaaatttaca
      901 taacagtgaa ctatgcagaa aattttactt ctggaccacg catttaacaa taatgtttgc
      961 ccgcaacaat aaagtatttg gtcgctttaa gacgttatca gttattttat gagaaggtgt
     1021 ttaccattcg tgcggccgca cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1081 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1141 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1201 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1261 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1321 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1381 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1441 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1501 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1561 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1621 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1681 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1741 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     1801 tgactgatcc ttcaactcag caaaagttcg atttattcaa caaagccacg ttgtgtctca
     1861 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     1921 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg
     1981 ctccaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     2041 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
     2101 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2161 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     2221 tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt
     2281 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2341 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgcctcgc
     2401 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     2461 taatggctgg cctgttgaac aagtctggaa agaaatgcat aagcttttgc cattctcacc
     2521 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2581 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2641 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     2701 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     2761 tttctaatca gaattggtta attggttgta acacgcggcc gcctctttcc tctacaaaag
     2821 tatgttaaat gtaggtagtt caaattgcgg tcacgtatat gccaacgttg ttaatatata
     2881 atgatccatt ggaacaatga agtttacaga agctcaaaat taatgcaaga ataattacgt
     2941 caagggagtc ataacaaact aagaaaacaa aggaatttga ctaagtttga gcgcaatgat
     3001 atatccatgc ttgatattct gattcataca tatcgtttct gtcatttcaa tggaagtctt
     3061 tatttatcaa ttaccacaac ctttagtatt gtgcttatta ttggtcaaaa ggagttcatg
     3121 cgctagtgat agtcaaccca tgaagtgatt agtggaacat agaaggggat aaatttccta
     3181 tcggataaca aagaaaaacc aggattattg ttttaccacc acattttcaa actttcccgc
     3241 tgaatttcct actaccctca tttgaaagaa cgctttccac tcggggtaat tctagcggtc
     3301 atactcatgt cttcgtcaga agacattgta ctgactgcag cagtactagt acgttgcttg
     3361 agacgtgcac agctgagacc cagacgagca ccaaataccc gctggactgt tgtgatatgc
     3421 catatatgca gtggtgttcg ccggcgctgg ttccggctgt cttgcttagt tgtgagtcgc
     3481 caggcagtca tcggtatgat ctgtacatga ttcgtcagtg tgagcaccac tgacgagcag
     3541 atttcgcggc cgcagtacag ctctaggtct ataaaaaaaa aaattggtta aagaaaaaat
     3601 atacaggttt gtatatagaa tcattaatta aaaagataat agtataaaac acatcagata
     3661 aacaaaagtt tgcccaataa aattatattt tccatcaacc agctacttgt acaactcatc
     3721 cataccacca gtagaatgtc taccttctga tctttcgtac tgttcaacaa cggtgtaatc
     3781 ttcgttgtga gaggtgatat ccaattttct atcaacgtta taagcaccag gcatttgaac
     3841 tggctttttg gccttgtaag tagtcttaaa atcagccaag tatctaccac cgtctttcaa
     3901 tctcaaggcc atcttaatat cacctttcaa aacaccatcc tctggatata atctttcagt
     3961 agaagcttcc catcccatag tctttttttg cataacagga ccatctggtg ggaaattagt
     4021 acctcttaac ttaaccttgt agatcaaggt gccatcttcc aatgaagtat cttgagtaac
     4081 agtaacggca ccaccatctt caaaattcat gactctttcc cacttgaaac cctctggaaa
     4141 tgattgctta taataatctg ggatatcagc tgggtgtttt gtaaaagctc ttgaaccgta
     4201 cataaattgt ggagacaaaa tatcccaaga gaatggcaaa ggaccacctt ttgtaacctt
     4261 caatttagca gtttgtgtac cttcgtatgg tctaccctca ccttcaccct caatttcaaa
     4321 ttcgtgaccg ttcatagaac cttccatgtg aaccttgaat ctcataaatt ctttgatgac
     4381 ggcttcacct ttagacacca ttttaattgt ttgtgtgtgt gtgttttcga aactaagttc
     4441 ttggggtttt aaaatcaata agaattaaaa tagaataact ataaaataag aagtgtaaag
     4501 gagttatagt tttatgagtt tatgatcgat agttgtcgtc tttatatatt tattaatcaa
     4561 acagggggat gatttgtgga aactggagac aacctttttt tcagtttttt ccaacccaga
     4621 gagttagaag gtaatagaag gtgtgagaaa atagacagac acgtaggtcg attgccttgt
     4681 gtcagctttt tgctccagtc agtttgcatc actccattga tttttttcag gaagccagat
     4741 taaaaagcgc gatccgagat atttagtgga tgctggaaaa gggaccgctc acccagacat
     4801 tcggaatgtg gcctgtggct gccgtaaccc gctcaatgag aagtcgggtt actaatggca
     4861 ggtgaaagat gaatgtataa acgcagaaag gcccacccga aggtgagcca gtgtgactct
     4921 agtagagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct ttcgactgag
     4981 cctttcgttt tatttgatgc ctggtcactt atacagttcg tccatgccgc cggtgctgtg
     5041 gcgaccctcg ctacgctcat attgctcaac cacggtataa tcctcgttgt ggctggtaat
     5101 atccagttta cggtcaacgt tgtacgcacc cggcatttgc accggtttct tcgctttata
     5161 ggtggtctta aaatccgcca ggtaacggcc accgtccttc agacgcagcg ccattttgat
     5221 atcacccttc agcacgccat cttccggata cagacgctcg gtgctcgctt cccaacccat
     5281 cgttttcttc tgcataaccg gaccatccgg cggaaagttg gtgccacgca gtttcacctt
     5341 gtaaatcagg gtgccatcct ccaggctggt gtcctgggta acggtcaccg cgccaccatc
     5401 ttcgaagttc ataacacgct cccatttaaa accttccggg aagctttgct tatagtaatc
     5461 cggaatgtcc gccgggtgtt tggtaaacgc acggctgcca tacatgaact gcgggctcag
     5521 gatgtcccag ctgaacggca gcgggccacc cttggtcact ttcagcttcg cggtctgggt
     5581 gccttcgtac ggacgaccct cgccttcacc ctcga
//
