LOCUS       sequence-369773-        5685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Markerless yeast integration Vector at Int.6 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 5685)
  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 5685)
  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: pMYT080
FEATURES             Location/Qualifiers
     source          1..5685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1133..1948)
                     /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(1133..1948)
                     /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     1856..1875
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1940..1959
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2173..2761
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2662..2681
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             3008..3016
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     RBS             3756..3767
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      4481..4552
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4568..4595
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 ctcacaacga agattacacc gttgttgaac agtacgaaag atcagaaggt agacattcta
       61 ctggtggtat ggatgagttg tacaagtagc tggttgatgg aaaatataat tttattgggc
      121 aaacttttgt ttatctgatg tgttttatac tattatcttt ttaattaatg attctatata
      181 caaacctgta tattttttct ttaaccaatt ttttttttta tagacctaga gctgtactgc
      241 ggccgcgaaa tctgctcgtc agtggtgctc acactgacga atcatgtaca gatcataccg
      301 atgactgcct ggcgactcac aactaagcaa gacagccgga accagcgccg gcgaacacca
      361 ctgcatatat ggcatatcac aacagtccag cgggtatttg gtgctcgtct gggtctcagc
      421 tgtagccgtc tcaagcatca gactagttcg actgcagctg atacaatgtc ttcgactgaa
      481 gacatgagta tcccctaatc tccaaaccga caaggccatt aattcccttc ggagggctcc
      541 cgatgggagg attcttagca tgctctttct agccaacagt atcactttca tcagaaaacg
      601 gaagcctcca aaaagctacg ctaaacgctt ttttgttttg gattatcgca gaaaattggt
      661 gacattcttg attaagaata aacggtgact agcaagaaaa ggacatgtca ggtaccagta
      721 aagtccgatt taagatcaaa tcagcgcctt taaagaatct tgaggtctta gtaatcccga
      781 tggaaggaaa gaggtaaaca aacgaataac ttaaccctta agatgattct atacatagca
      841 tgatctatcc aaggaacgac cgacgccctt aaaagggttc tgctcagatt tcgttaaata
      901 cccggaccat cgtagatgat tgtttttact ctcccctaac gatgtactat gccataataa
      961 gcattttttt tttctttctt cactgcggct ctgctttgaa aacactacat ctaaggcgct
     1021 gacgccaacg cccagactaa atcatcatat aatatcaaac gcccaacaaa atattagaaa
     1081 tagctcaaca atagagcgcg gccgcgtgtt acaaccaatt aaccaattct gattagaaaa
     1141 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     1201 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     1261 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     1321 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     1381 gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac
     1441 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     1501 cgaggcgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     1561 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     1621 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     1681 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     1741 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     1801 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     1861 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctggagc
     1921 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     1981 acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
     2041 gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagtcat
     2101 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2161 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2221 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2281 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2341 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2401 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2461 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2521 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2581 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2641 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2701 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2761 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2821 gttctttcct gcgttatccc ctgattctgt ggataaccgt gcggccgctt aaaagccctt
     2881 attcaattga agaaatcccc tgacctctgc aaaaaatcta tcgtttattt tatttctttg
     2941 tcttacgtaa caaatttatt ttctgggatg atgtcaattc tatttggcac gcgagtcatc
     3001 gtctatataa ggaggtaacg aggtccttgt atatttgatg atcctcacta tcaacacgtc
     3061 gagggacggt tggtgacgaa ctcaacttat gttatagttt tggaccactc ttttccttga
     3121 aaaattacag gcattatacg cagtcctggg gcaggagcgc ttcgatcatt agcataacag
     3181 gatcacggtt atcacctgtt ttgttataat tcatggcctt taggtcatga cgtaaaataa
     3241 tcggagtgaa aaaatgaaat caaaggctat catttttagg ggaactaaag cacaaggatt
     3301 tgaaggcgta gtgaaataat gaataataga taatctatgg accgaacaca gacgtattta
     3361 aagacaatca caccccaatc cctagtagga attcgtactc tagagtcact gatgagacgt
     3421 acgaacgtga gaccctgtgg tcaaaagtgc gttggctggt agagccacaa acagccggta
     3481 caagcaacga tctccaggac catctgaatc atgcgcggat gacacgaact cacgacggcg
     3541 atcacagaca ttaacccaca gtacagacac tgcgacaacg tggcaattcg tcgcaatacg
     3601 cggccgcgaa agtgaaacgt gatttcatgc gtcattttga acattttgta aatcttattt
     3661 aataatgtgt gcggcaattc acatttaatt tatgaatgtt ttcttaacat cgcggcaact
     3721 caagaaacgg caggttcgga tcttagctac tagagaaaga ggagaaatac tagatggtta
     3781 gcaaaggcga ggcggttatc aaggagttta tgcgttttaa ggttcacatg gagggtagca
     3841 tgaacggtca cgagttcgag atcgagggtg aaggcgaggg tcgtccgtac gaaggcaccc
     3901 agaccgcgaa gctgaaagtg accaagggtg gcccgctgcc gttcagctgg gacatcctga
     3961 gcccgcagtt catgtatggc agccgtgcgt ttaccaaaca cccggcggac attccggatt
     4021 actataagca aagcttcccg gaaggtttta aatgggagcg tgttatgaac ttcgaagatg
     4081 gtggcgcggt gaccgttacc caggacacca gcctggagga tggcaccctg atttacaagg
     4141 tgaaactgcg tggcaccaac tttccgccgg atggtccggt tatgcagaag aaaacgatgg
     4201 gttgggaagc gagcaccgag cgtctgtatc cggaagatgg cgtgctgaag ggtgatatca
     4261 aaatggcgct gcgtctgaag gacggtggcc gttacctggc ggattttaag accacctata
     4321 aagcgaagaa accggtgcaa atgccgggtg cgtacaacgt tgaccgtaaa ctggatatta
     4381 ccagccacaa cgaggattat accgtggttg agcaatatga gcgtagcgag ggtcgccaca
     4441 gcaccggcgg catggacgaa ctgtataagt gaccaggcat caaataaaac gaaaggctca
     4501 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag
     4561 tcacactggc tcaccttcgg gtgggccttt ctgcgtttat acattcatct ttcacctgcc
     4621 attagtaacc cgacttctca ttgagcgggt tacggcagcc acaggccaca ttccgaatgt
     4681 ctgggtgagc ggtccctttt ccagcatcca ctaaatatct cggatcgcgc tttttaatct
     4741 ggcttcctga aaaaaatcaa tggagtgatg caaactgact ggagcaaaaa gctgacacaa
     4801 ggcaatcgac ctacgtgtct gtctattttc tcacaccttc tattaccttc taactctctg
     4861 ggttggaaaa aactgaaaaa aaggttgtct ccagtttcca caaatcatcc ccctgtttga
     4921 ttaataaata tataaagacg acaactatcg atcataaact cataaaacta taactccttt
     4981 acacttctta ttttatagtt attctatttt aattcttatt gattttaaaa ccccaagaac
     5041 ttagtttcga aaacacacac acacaaacaa ttaaaatggt gtctaaaggt gaagccgtca
     5101 tcaaagaatt tatgagattc aaggttcaca tggaaggttc tatgaacggt cacgaatttg
     5161 aaattgaggg tgaaggtgag ggtagaccat acgaaggtac acaaactgct aaattgaagg
     5221 ttacaaaagg tggtcctttg ccattctctt gggatatttt gtctccacaa tttatgtacg
     5281 gttcaagagc ttttacaaaa cacccagctg atatcccaga ttattataag caatcatttc
     5341 cagagggttt caagtgggaa agagtcatga attttgaaga tggtggtgcc gttactgtta
     5401 ctcaagatac ttcattggaa gatggcacct tgatctacaa ggttaagtta agaggtacta
     5461 atttcccacc agatggtcct gttatgcaaa aaaagactat gggatgggaa gcttctactg
     5521 aaagattata tccagaggat ggtgttttga aaggtgatat taagatggcc ttgagattga
     5581 aagacggtgg tagatacttg gctgatttta agactactta caaggccaaa aagccagttc
     5641 aaatgcctgg tgcttataac gttgatagaa aattggatat cacct
//
