LOCUS       sequence-369777-        5612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Markerless yeast integration Vector at Int.7 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 5612)
  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 5612)
  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: pMYT081
FEATURES             Location/Qualifiers
     source          1..5612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             450..461
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      complement(1461..2049)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1541..1560)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2263..2282)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2274..3089
                     /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             2274..3089
                     /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(2347..2366)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(5234..5261)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(5277..5348)
                     /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 gttgctacgt ctcatcagcg tatctagatg acgaattcgg cacggtatgt
      841 tttaaggccc gcggcaccac attttggcat acgtcgggac attggcggct gaatagatta
      901 tacccgtcgc ccttttttca tctctccttt tctttttgtc gtaagcaaat tacagaattg
      961 aatatttgaa taaccctaaa aaaaatgcca ctcccctaaa aacaaaccag attaagttgc
     1021 aagaaataga ctgattggaa catacgggtc ctattcagtg ctcaataaaa attaaaataa
     1081 tgtgatcaaa caacccgaga ctattcttga attagcatta gccactataa attttaacat
     1141 ttttcagagt ctttcgtgcg gggaccgtaa atattagcat tgaccttgtc aatgaagtaa
     1201 ttgtctttga ttttaaccgt gttaaagatt tcgcaaaaga gaaagcttgc tggcttggag
     1261 cagtaacttt ttttgatttt tttttacaag cacacatgct tttgccttta gaatattacg
     1321 gctcctcatg tggatagtaa aaacttgacc gttgcggccg cacggttatc cacagaatca
     1381 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1441 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     1501 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1561 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1621 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1681 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1741 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1801 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1861 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1921 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1981 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2041 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     2101 tcacgttaag ggattttggt catgactgat ccttcaactc agcaaaagtt cgatttattc
     2161 aacaaagcca cgttgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     2221 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     2281 atattcaacg ggaaacgtct tgctccaggc cgcgattaaa ttccaacatg gatgctgatt
     2341 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat
     2401 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     2461 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     2521 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     2581 ggaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     2641 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     2701 gcgatcgcgt atttcgcctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     2761 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     2821 ataagctttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     2881 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     2941 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     3001 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     3061 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacacgcgg
     3121 ccgctacgcc ccagctttac tacgtggatg atacagtaag cacatacata ttgacggcat
     3181 ctaatcgttc taaacggaaa aataggcgct gatttgatcc acttcggtta ggtaaccgct
     3241 gatgccaacg atctagggcc cttccatttt aggaaaaagg gtatatactc tgatgccctg
     3301 tctggccgaa cattcgtata tgtggatgta tcgtgatccg ttcgtattca ataaaagtgt
     3361 aacgatctca tctcaagtca aaagtgagaa gggaataact aaaaaagaaa aactgatatt
     3421 ttgacgtgac gtggtttatt agcaagtatg tttcaaccat tctcagcgta aaccctatca
     3481 caataacgcc tctatgaaaa gtttaacatt ttgtctatct cggttcaatc agttgaatag
     3541 cgaaccttat agcaaatagc attttcaata aaaatgtgac atacagactt ccacttttct
     3601 tgcttttctc atgctctgtg tgatgctact gttaaatcct ccacctctta cccgactcat
     3661 gtcttctagc gaagacattg taatgcctgc aggtcaacta gttcgatgct tgagacgctg
     3721 acagctgaga cccagacgag caccaaatac ccgctggact gttgtgatat gccatatatg
     3781 cagtggtgtt cgccggcgct ggttccggct gtcttgctta gttgtgagtc gccaggcagt
     3841 catcggtatg atctgtacat gattcgtcag tgtgagcacc actgacgagc agatttcgcg
     3901 gccgcagtac agctctaggt ctataaaaaa aaaaattggt taaagaaaaa atatacaggt
     3961 ttgtatatag aatcattaat taaaaagata atagtataaa acacatcaga taaacaaaag
     4021 tttgcccaat aaaattatat tttccatcaa ccagctactt gtacaactca tccataccac
     4081 cagtagaatg tctaccttct gatctttcgt actgttcaac aacggtgtaa tcttcgttgt
     4141 gagaggtgat atccaatttt ctatcaacgt tataagcacc aggcatttga actggctttt
     4201 tggccttgta agtagtctta aaatcagcca agtatctacc accgtctttc aatctcaagg
     4261 ccatcttaat atcacctttc aaaacaccat cctctggata taatctttca gtagaagctt
     4321 cccatcccat agtctttttt tgcataacag gaccatctgg tgggaaatta gtacctctta
     4381 acttaacctt gtagatcaag gtgccatctt ccaatgaagt atcttgagta acagtaacgg
     4441 caccaccatc ttcaaaattc atgactcttt cccacttgaa accctctgga aatgattgct
     4501 tataataatc tgggatatca gctgggtgtt ttgtaaaagc tcttgaaccg tacataaatt
     4561 gtggagacaa aatatcccaa gagaatggca aaggaccacc ttttgtaacc ttcaatttag
     4621 cagtttgtgt accttcgtat ggtctaccct caccttcacc ctcaatttca aattcgtgac
     4681 cgttcataga accttccatg tgaaccttga atctcataaa ttctttgatg acggcttcac
     4741 ctttagacac cattttaatt gtttgtgtgt gtgtgttttc gaaactaagt tcttggggtt
     4801 ttaaaatcaa taagaattaa aatagaataa ctataaaata agaagtgtaa aggagttata
     4861 gttttatgag tttatgatcg atagttgtcg tctttatata tttattaatc aaacaggggg
     4921 atgatttgtg gaaactggag acaacctttt tttcagtttt ttccaaccca gagagttaga
     4981 aggtaataga aggtgtgaga aaatagacag acacgtaggt cgattgcctt gtgtcagctt
     5041 tttgctccag tcagtttgca tcactccatt gatttttttc aggaagccag attaaaaagc
     5101 gcgatccgag atatttagtg gatgctggaa aagggaccgc tcacccagac attcggaatg
     5161 tggcctgtgg ctgccgtaac ccgctcaatg agaagtcggg ttactaatgg caggtgaaag
     5221 atgaatgtat aaacgcagaa aggcccaccc gaaggtgagc cagtgtgact ctagtagaga
     5281 gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt
     5341 tttatttgat gcctggtcac ttatacagtt cgtccatgcc gccggtgctg tggcgaccct
     5401 cgctacgctc atattgctca accacggtat aatcctcgtt gtggctggta atatccagtt
     5461 tacggtcaac gttgtacgca cccggcattt gcaccggttt cttcgcttta taggtggtct
     5521 taaaatccgc caggtaacgg ccaccgtcct tcagacgcag cgccattttg atatcaccct
     5581 tcagcacgcc atcttccgga tacagacgct cg
//
