LOCUS       sequence-369761-        5632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Markerless yeast integration Vector at Int.5 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 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: pMYT079
FEATURES             Location/Qualifiers
     source          1..5632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             450..461
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      complement(1479..2067)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1559..1578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2281..2300)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2292..3107
                     /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             2292..3107
                     /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(2365..2384)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(5254..5281)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(5297..5368)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtgctcgctt cccaacccat cgttttcttc tgcataaccg gaccatccgg cggaaagttg
       61 gtgccacgca gtttcacctt gtaaatcagg gtgccatcct ccaggctggt gtcctgggta
      121 acggtcaccg cgccaccatc ttcgaagttc ataacacgct cccatttaaa accttccggg
      181 aagctttgct tatagtaatc cggaatgtcc gccgggtgtt tggtaaacgc acggctgcca
      241 tacatgaact gcgggctcag gatgtcccag ctgaacggca gcgggccacc cttggtcact
      301 ttcagcttcg cggtctgggt gccttcgtac ggacgaccct cgccttcacc ctcgatctcg
      361 aactcgtgac cgttcatgct accctccatg tgaaccttaa aacgcataaa ctccttgata
      421 accgcctcgc ctttgctaac catctagtat ttctcctctt tctctagtag ctaagatccg
      481 aacctgccgt ttcttgagtt gccgcgatgt taagaaaaca ttcataaatt aaatgtgaat
      541 tgccgcacac attattaaat aagatttaca aaatgttcaa aatgacgcat gaaatcacgt
      601 ttcactttcg cggccgcgta ttgcgacgaa ttgccacgtt gtcgcagtgt ctgtactgtg
      661 ggttaatgtc tgtgatcgcc gtcgtgagtt cgtgtcatcc gcgcatgatt cagatggtcc
      721 tggagatcgt tgcttgtacc ggctgtttgt ggctctacca gccaacgcac ttttgaccac
      781 agggtctcac gtttgaccgt ctcatcaggt actctagata gcgaattcag atgcccactt
      841 ctgataggat atgtacctca attgcagata gcatcaaatt tagactacgt catcaaccaa
      901 attacaccct atgaaaaaca taagattata agcatgcgtt tgtgtaaagg tcataccatt
      961 ttatacgttg acgcaatggt gcaaatagaa aggtttacga aagctcttat tccgcttagc
     1021 gcaattctat tgttaatttc aagtaaaaag ataaatatct cacgtcctta tattatcttt
     1081 gattaaaatt tcacgatcaa tgtaacgaaa aaaggtcgca aatatttctt tttcatcact
     1141 ctgattaaac aatggcatca tgaaacgctc attggatcgt tacacgttct cattgtacat
     1201 atgtatgatt tagaacagct tgttcgcctg atctccacct tataccctca tcttaccgca
     1261 gtgcggtttt acgcctcatg ttatttttgc cagctttaat aacaacatca gtaatattac
     1321 gttattggat cttccactcc ggttcgagga aaaaaagaga ggaggagaaa cgcggccgca
     1381 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1441 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1501 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1561 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1621 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     1681 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1741 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1801 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     1861 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     1921 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1981 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2041 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2101 gctcagtgga acgaaaactc acgttaaggg attttggtca tgactgatcc ttcaactcag
     2161 caaaagttcg atttattcaa caaagccacg ttgtgtctca aaatctctga tgttacattg
     2221 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     2281 caaggggtgt tatgagccat attcaacggg aaacgtcttg ctccaggccg cgattaaatt
     2341 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     2401 gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     2461 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     2521 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     2581 tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat cctgattcag
     2641 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     2701 gtaattgtcc ttttaacagc gatcgcgtat ttcgcctcgc tcaggcgcaa tcacgaatga
     2761 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     2821 aagtctggaa agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg
     2881 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     2941 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     3001 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     3061 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta
     3121 attggttgta acacgcggcc gcaacttttg atgattgctg ggaaacggtt tggtaaactg
     3181 ctgaagcaat tgttgtacac aatacttctg cttttggccc catctggatt tttcatttcc
     3241 gcttttgcct caggtgcaac aataactatc tttgagcatc gtaccaccaa tacaaaagga
     3301 tatgatcgtt atcatacaga atttttcatg gattttgcat gtttcggtat aactttacgt
     3361 aggtgaatta ttgaagggct ttgtgatagg tgtggatgta cctgccgtgc gttgatgacc
     3421 ttcaactgat cggaacaaat tgctattaca tgtaaaaatc tcatgtatga gcgaatcttt
     3481 ttgtgttata ggcaatcacc tagaaaagct tggaaattgg tcctgagttt tttgttgatg
     3541 gtaagtgcat ttttttagta tgaaaggttt acgatattag ctgctaagtt ctcccttgcc
     3601 ctagtgaatt acgttttttt cgattaaaag ggttggtccc gaaattgacc tcttcgcaga
     3661 cagaggaatg aatcactcat gtcttcctag gaagacattg tatacgctgc agctgaacta
     3721 gtgctatgct tgagacgcgt acagctgaga cccagacgag caccaaatac ccgctggact
     3781 gttgtgatat gccatatatg cagtggtgtt cgccggcgct ggttccggct gtcttgctta
     3841 gttgtgagtc gccaggcagt catcggtatg atctgtacat gattcgtcag tgtgagcacc
     3901 actgacgagc agatttcgcg gccgcagtac agctctaggt ctataaaaaa aaaaattggt
     3961 taaagaaaaa atatacaggt ttgtatatag aatcattaat taaaaagata atagtataaa
     4021 acacatcaga taaacaaaag tttgcccaat aaaattatat tttccatcaa ccagctactt
     4081 gtacaactca tccataccac cagtagaatg tctaccttct gatctttcgt actgttcaac
     4141 aacggtgtaa tcttcgttgt gagaggtgat atccaatttt ctatcaacgt tataagcacc
     4201 aggcatttga actggctttt tggccttgta agtagtctta aaatcagcca agtatctacc
     4261 accgtctttc aatctcaagg ccatcttaat atcacctttc aaaacaccat cctctggata
     4321 taatctttca gtagaagctt cccatcccat agtctttttt tgcataacag gaccatctgg
     4381 tgggaaatta gtacctctta acttaacctt gtagatcaag gtgccatctt ccaatgaagt
     4441 atcttgagta acagtaacgg caccaccatc ttcaaaattc atgactcttt cccacttgaa
     4501 accctctgga aatgattgct tataataatc tgggatatca gctgggtgtt ttgtaaaagc
     4561 tcttgaaccg tacataaatt gtggagacaa aatatcccaa gagaatggca aaggaccacc
     4621 ttttgtaacc ttcaatttag cagtttgtgt accttcgtat ggtctaccct caccttcacc
     4681 ctcaatttca aattcgtgac cgttcataga accttccatg tgaaccttga atctcataaa
     4741 ttctttgatg acggcttcac ctttagacac cattttaatt gtttgtgtgt gtgtgttttc
     4801 gaaactaagt tcttggggtt ttaaaatcaa taagaattaa aatagaataa ctataaaata
     4861 agaagtgtaa aggagttata gttttatgag tttatgatcg atagttgtcg tctttatata
     4921 tttattaatc aaacaggggg atgatttgtg gaaactggag acaacctttt tttcagtttt
     4981 ttccaaccca gagagttaga aggtaataga aggtgtgaga aaatagacag acacgtaggt
     5041 cgattgcctt gtgtcagctt tttgctccag tcagtttgca tcactccatt gatttttttc
     5101 aggaagccag attaaaaagc gcgatccgag atatttagtg gatgctggaa aagggaccgc
     5161 tcacccagac attcggaatg tggcctgtgg ctgccgtaac ccgctcaatg agaagtcggg
     5221 ttactaatgg caggtgaaag atgaatgtat aaacgcagaa aggcccaccc gaaggtgagc
     5281 cagtgtgact ctagtagaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
     5341 ctttcgactg agcctttcgt tttatttgat gcctggtcac ttatacagtt cgtccatgcc
     5401 gccggtgctg tggcgaccct cgctacgctc atattgctca accacggtat aatcctcgtt
     5461 gtggctggta atatccagtt tacggtcaac gttgtacgca cccggcattt gcaccggttt
     5521 cttcgcttta taggtggtct taaaatccgc caggtaacgg ccaccgtcct tcagacgcag
     5581 cgccattttg atatcaccct tcagcacgcc atcttccgga tacagacgct cg
//
