LOCUS       sequence-369788-        2986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Markerless yeast integration Spacer at Int.6 locus to used in the 
            Ellis MYT system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2986)
  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 2986)
  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: pMYT090
FEATURES             Location/Qualifiers
     source          1..2986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1165..1173
                     /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)"
     rep_origin      complement(1420..2008)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1500..1519)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2222..2241)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             join(2233..2986,1..62)
                     /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             join(2233..2986,1..62)
                     /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(2306..2325)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
       61 aatcagaatt ggttaattgg ttgtaacacg cggccgcgct ctattgttga gctatttcta
      121 atattttgtt gggcgtttga tattatatga tgatttagtc tgggcgttgg cgtcagcgcc
      181 ttagatgtag tgttttcaaa gcagagccgc agtgaagaaa gaaaaaaaaa atgcttatta
      241 tggcatagta catcgttagg ggagagtaaa aacaatcatc tacgatggtc cgggtattta
      301 acgaaatctg agcagaaccc ttttaagggc gtcggtcgtt ccttggatag atcatgctat
      361 gtatagaatc atcttaaggg ttaagttatt cgtttgttta cctctttcct tccatcggga
      421 ttactaagac ctcaagattc tttaaaggcg ctgatttgat cttaaatcgg actttactgg
      481 tacctgacat gtccttttct tgctagtcac cgtttattct taatcaagaa tgtcaccaat
      541 tttctgcgat aatccaaaac aaaaaagcgt ttagcgtagc tttttggagg cttccgtttt
      601 ctgatgaaag tgatactgtt ggctagaaag agcatgctaa gaatcctccc atcgggagcc
      661 ctccgaaggg aattaatggc cttgtcggtt tggagattag gggatactca tgtcttcagt
      721 cgaagacatt gtatcagctg cagtcgaact agtccgggtc ggggtatcct caagtgtcta
      781 gagtacgaat tcctactagg gattggggtg tgattgtctt taaatacgtc tgtgttcggt
      841 ccatagatta tctattattc attatttcac tacgccttca aatccttgtg ctttagttcc
      901 cctaaaaatg atagcctttg atttcatttt ttcactccga ttattttacg tcatgaccta
      961 aaggccatga attataacaa aacaggtgat aaccgtgatc ctgttatgct aatgatcgaa
     1021 gcgctcctgc cccaggactg cgtataatgc ctgtaatttt tcaaggaaaa gagtggtcca
     1081 aaactataac ataagttgag ttcgtcacca accgtccctc gacgtgttga tagtgaggat
     1141 catcaaatat acaaggacct cgttacctcc ttatatagac gatgactcgc gtgccaaata
     1201 gaattgacat catcccagaa aataaatttg ttacgtaaga caaagaaata aaataaacga
     1261 tagatttttt gcagaggtca ggggatttct tcaattgaat aagggctttt aagcggccgc
     1321 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1381 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1441 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1501 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1561 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1621 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1681 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1741 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1801 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1861 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1921 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1981 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2041 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgactgatc cttcaactca
     2101 gcaaaagttc gatttattca acaaagccac gttgtgtctc aaaatctctg atgttacatt
     2161 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     2221 acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctccaggcc gcgattaaat
     2281 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     2341 ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     2401 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     2461 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     2521 ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata tcctgattca
     2581 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     2641 tgtaattgtc cttttaacag cgatcgcgta tttcgcctcg ctcaggcgca atcacgaatg
     2701 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     2761 caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt cgtcactcat
     2821 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     2881 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     2941 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatg
//
