LOCUS       sequence-369380-        3276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes HygR as a yeast Vector selectable marker to be used in the 
            Ellis MYT system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3276)
  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 3276)
  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: pMYT037
FEATURES             Location/Qualifiers
     source          1..3276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      247..281
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     gene            complement(298..1872)
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     terminator      complement(298..495)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(500..1528)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             complement(500..1528)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     rep_origin      complement(2023..2611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2103..2122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2793..3276,1..176)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2859..2878)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca
       61 aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg
      121 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttg
      181 atatcgagct cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg
      241 gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgaaga catactccag
      301 tatagcgacc agcattcaca tacgattgac gcatgatatt actttctgcg cacttaactt
      361 cgcatctggg cagatgatgt cgaggcgaaa aaaaatataa atcacgctaa catttgatta
      421 aaatagaaca actacaatat aaaaaaacta tacaaatgac aagttcttga aaacaagaat
      481 ctttttattg tcagttactt tattcctttg ccctcggacg agtgctgggg cgtcggtttc
      541 cactatcggc gagtacttct acacagccat cggtccagac ggccgcgctt ctgcgggcga
      601 tttgtgtacg cccgacagtc ccggctccgg atcggacgat tgcgtcgcat cgaccctgcg
      661 cccaagctgc atcatcgaaa ttgccgtcaa ccaagctctg atagagttgg tcaagaccaa
      721 tgcggagcat atacgcccgg agccgcggcg atcctgcaag ctccggatgc ctccgctcga
      781 agtagcgcgt ctgctgctcc atacaagcca accacggcct ccagaagaag atgttggcga
      841 cctcgtattg ggaatccccg aacatcgcct cgctccagtc aatgaccgct gttatgcggc
      901 cattgtccgt caggacattg ttggagccga aatccgcgtg cacgaggtgc cggacttcgg
      961 ggcagtcctc ggcccaaagc atcagctcat cgagagcctg cgcgacggac gcactgacgg
     1021 tgtcgtccat cacagtttgc cagtgataca catggggatc agcaatcgcg catatgaaat
     1081 cacgccatgt agtgtattga ccgattcctt gcggtccgaa tgggccgaac ccgctcgtct
     1141 ggctaagatc ggccgcagcg atcgcatcca tggcctccgc gaccggttgc agaacagcgg
     1201 gcagttcggt ttcaggcagg tcttgcaacg tgacaccctg tgcacggcgg gagatgcaat
     1261 aggtcaggct ctcgctaaat tccccaatgt caagcacttc cggaatcggg agcgcggccg
     1321 atgcaaagtg ccgataaaca taacgatctt tgtagaaacc atcggcgcag ctatttaccc
     1381 gcaggacata tccacgccct cctacatcga agctgaaagc acgagattct tcgccctccg
     1441 agagctgcat caggtcggac acgctgtcga acttttcgat cagaaacttc tcgacagacg
     1501 tcgcggtgag ttcaggcttt ttacccattt ttgtttatgt tcggatgtga tgtgagaact
     1561 gtatcctagc aagattttaa aaggaagtat atgaaagaag aacctcagtg gcaaatccta
     1621 accttttata tttctctaca ggggcgcggc gtggggacaa ttcaacgcgt ctgtgagggg
     1681 agcgtttccc tgctcgcagg tctggagcga ggagccgtaa tttttgcttc gcgccgtgcg
     1741 gccatcaaaa tgtatggatg caaatgatta tacatgggga tgtatgggct aaatgtacgg
     1801 gcgacagtca catcatgccc ctgagctgcg cacgtcaaga ctgtcaagga gggtattctg
     1861 ggcctccatg tcgctggccg ggtgacccgg cggggacgag gcaagctaaa ctgtaatgtc
     1921 ttcacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1981 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2041 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2101 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2161 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2221 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2281 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2341 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2401 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2461 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2521 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2581 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2641 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgattg atcgggcacg
     2701 taagaggttc caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt
     2761 tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca
     2821 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
     2881 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
     2941 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
     3001 atccggaatt tcgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc
     3061 cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc
     3121 acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa
     3181 acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtttca gccaatccct
     3241 gggtgagttt caccagtttt gatttaaacg tggcca
//
