LOCUS       sequence-369729-        5651 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Markerless yeast integration Vector at Int.1 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 5651)
  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 5651)
  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: pMYT075
FEATURES             Location/Qualifiers
     source          1..5651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1484..1511)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(1527..1598)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             2312..2323
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      complement(3337..3925)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3417..3436)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4139..4158)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4150..4965
                     /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             4150..4965
                     /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(4223..4242)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ccgctggact gttgtgatat gccatatatg cagtggtgtt cgccggcgct ggttccggct
       61 gtcttgctta gttgtgagtc gccaggcagt catcggtatg atctgtacat gattcgtcag
      121 tgtgagcacc actgacgagc agatttcgcg gccgcagtac agctctaggt ctataaaaaa
      181 aaaaattggt taaagaaaaa atatacaggt ttgtatatag aatcattaat taaaaagata
      241 atagtataaa acacatcaga taaacaaaag tttgcccaat aaaattatat tttccatcaa
      301 ccagctactt gtacaactca tccataccac cagtagaatg tctaccttct gatctttcgt
      361 actgttcaac aacggtgtaa tcttcgttgt gagaggtgat atccaatttt ctatcaacgt
      421 tataagcacc aggcatttga actggctttt tggccttgta agtagtctta aaatcagcca
      481 agtatctacc accgtctttc aatctcaagg ccatcttaat atcacctttc aaaacaccat
      541 cctctggata taatctttca gtagaagctt cccatcccat agtctttttt tgcataacag
      601 gaccatctgg tgggaaatta gtacctctta acttaacctt gtagatcaag gtgccatctt
      661 ccaatgaagt atcttgagta acagtaacgg caccaccatc ttcaaaattc atgactcttt
      721 cccacttgaa accctctgga aatgattgct tataataatc tgggatatca gctgggtgtt
      781 ttgtaaaagc tcttgaaccg tacataaatt gtggagacaa aatatcccaa gagaatggca
      841 aaggaccacc ttttgtaacc ttcaatttag cagtttgtgt accttcgtat ggtctaccct
      901 caccttcacc ctcaatttca aattcgtgac cgttcataga accttccatg tgaaccttga
      961 atctcataaa ttctttgatg acggcttcac ctttagacac cattttaatt gtttgtgtgt
     1021 gtgtgttttc gaaactaagt tcttggggtt ttaaaatcaa taagaattaa aatagaataa
     1081 ctataaaata agaagtgtaa aggagttata gttttatgag tttatgatcg atagttgtcg
     1141 tctttatata tttattaatc aaacaggggg atgatttgtg gaaactggag acaacctttt
     1201 tttcagtttt ttccaaccca gagagttaga aggtaataga aggtgtgaga aaatagacag
     1261 acacgtaggt cgattgcctt gtgtcagctt tttgctccag tcagtttgca tcactccatt
     1321 gatttttttc aggaagccag attaaaaagc gcgatccgag atatttagtg gatgctggaa
     1381 aagggaccgc tcacccagac attcggaatg tggcctgtgg ctgccgtaac ccgctcaatg
     1441 agaagtcggg ttactaatgg caggtgaaag atgaatgtat aaacgcagaa aggcccaccc
     1501 gaaggtgagc cagtgtgact ctagtagaga gcgttcaccg acaaacaaca gataaaacga
     1561 aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctggtcac ttatacagtt
     1621 cgtccatgcc gccggtgctg tggcgaccct cgctacgctc atattgctca accacggtat
     1681 aatcctcgtt gtggctggta atatccagtt tacggtcaac gttgtacgca cccggcattt
     1741 gcaccggttt cttcgcttta taggtggtct taaaatccgc caggtaacgg ccaccgtcct
     1801 tcagacgcag cgccattttg atatcaccct tcagcacgcc atcttccgga tacagacgct
     1861 cggtgctcgc ttcccaaccc atcgttttct tctgcataac cggaccatcc ggcggaaagt
     1921 tggtgccacg cagtttcacc ttgtaaatca gggtgccatc ctccaggctg gtgtcctggg
     1981 taacggtcac cgcgccacca tcttcgaagt tcataacacg ctcccattta aaaccttccg
     2041 ggaagctttg cttatagtaa tccggaatgt ccgccgggtg tttggtaaac gcacggctgc
     2101 catacatgaa ctgcgggctc aggatgtccc agctgaacgg cagcgggcca cccttggtca
     2161 ctttcagctt cgcggtctgg gtgccttcgt acggacgacc ctcgccttca ccctcgatct
     2221 cgaactcgtg accgttcatg ctaccctcca tgtgaacctt aaaacgcata aactccttga
     2281 taaccgcctc gcctttgcta accatctagt atttctcctc tttctctagt agctaagatc
     2341 cgaacctgcc gtttcttgag ttgccgcgat gttaagaaaa cattcataaa ttaaatgtga
     2401 attgccgcac acattattaa ataagattta caaaatgttc aaaatgacgc atgaaatcac
     2461 gtttcacttt cgcggccgcg tattgcgacg aattgccacg ttgtcgcagt gtctgtactg
     2521 tgggttaatg tctgtgatcg ccgtcgtgag ttcgtgtcat ccgcgcatga ttcagatggt
     2581 cctggagatc gttgcttgta ccggctgttt gtggctctac cagccaacgc acttttgacc
     2641 acagggtctc acgtttctcc gtctcatcag agtctctaga ttgggaattc agtcgtggat
     2701 gtgatgggta atgaggatcg aatggacaaa agtgttttca ggcatgggag aatgggatgc
     2761 ttatagagga gtttaaccat gaaattaaat gattctccag caagaaaaaa aaaactcctg
     2821 actcagttca tgtggcggga aatttctctc attgtctccc cctgcggcgc tactaatctt
     2881 aaaggtagtg gtggcttcga cattatatcg ttggttcagg ttgtccgatc actaatgatc
     2941 tgttacttta ttgctatgtt atgtgtcaaa aggtgatatc cccattttta ctgatcgcag
     3001 taatatttta acctgccatg gttaaattgt ttattgtcat tgagttgagc actttttcat
     3061 tgcactgtag ttgttgcgta tttcagactt actcttatag tgacgctaac aggaggtctg
     3121 gctcttaata caaggacgtc gtaatattct gttcaccgga ctcgaaagca agaaggaaaa
     3181 aagcacattg tagccttttc aaaaacctat cttttggtac taatttatgg cggccgcacg
     3241 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3301 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3361 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3421 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3481 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3541 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3601 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3661 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3721 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3781 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3841 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3901 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3961 tcagtggaac gaaaactcac gttaagggat tttggtcatg actgatcctt caactcagca
     4021 aaagttcgat ttattcaaca aagccacgtt gtgtctcaaa atctctgatg ttacattgca
     4081 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     4141 aggggtgtta tgagccatat tcaacgggaa acgtcttgct ccaggccgcg attaaattcc
     4201 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     4261 gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     4321 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     4381 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     4441 accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc tgattcaggt
     4501 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     4561 aattgtcctt ttaacagcga tcgcgtattt cgcctcgctc aggcgcaatc acgaatgaat
     4621 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     4681 gtctggaaag aaatgcataa gcttttgcca ttctcaccgg attcagtcgt cactcatggt
     4741 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     4801 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     4861 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     4921 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     4981 tggttgtaac acgcggccgc aacagtgggg aaatagttta cgagttttgt tcaagttaaa
     5041 agcatttttt gttgtgaaga gaataaactt gatttcgaag gagatcaaat tcgaaataat
     5101 ctgcgaattt attatgatcg actaggcaac aaagacgcag ttttctatta aatctgtgcg
     5161 tttgagtaac ttaactgact ataatatcgc aatcttaaca tcattaaaca atgtaaaaga
     5221 ggtcatacgc tataagaagt tgatttgatc ccccgtggat tttgaaaaat gtcaagatca
     5281 gcatattgac ccatgttttt ccgctcaatc cagtctatca atcatttttc aacaaggata
     5341 accataacat tcacccacca tgctttggtt cataagatat acggggagat cagcaactct
     5401 tcaacttatt gaacatgttg agagcaggat taaataatca atgtcctgtg attgcaacat
     5461 ctatcatgcc tgacgtttct cgaacgatac aatctacaat ttaatgagcc attttagcat
     5521 attaacgagg gctgccgttt ccttcgtgct tagacactca tgtcttccag tgaagacatt
     5581 gtaacgtctg caggtacact agttagctgc ttgagacgat gccagctgag acccagacga
     5641 gcaccaaata c
//
