LOCUS       sequence-309854-        4682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low-copy expression vector for K. marxianus, histidine selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4682)
  AUTHORS   Rajkumar AS, Morrissey JP
  TITLE     Rational engineering of Kluyveromyces marxianus to create a chassis 
            for the production of aromatic products.
  JOURNAL   Microb Cell Fact. 2020 Nov 11;19(1):207. doi: 
            10.1186/s12934-020-01461-7.
  PUBMED    33176787
REFERENCE   2  (bases 1 to 4682)
  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: "Microb Cell
            Fact."; date: "2020-11-11"; volume: "19(1):207. doi"; pages: " 
            10.1186/s12934-020-01461-7"
COMMENT     Sequence Label: pMTU-DO-HIS
FEATURES             Location/Qualifiers
     source          1..4682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             334..345
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             352..1065
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1077..1148
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1164..1191
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1452..1639
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             1640..2302
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     CDS             complement(2947..3762)
                     /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             complement(2947..3762)
                     /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     3670..3689
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3754..3773
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3987..4575
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4476..4495
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctggaatt cgcatctaga ctgatgagac gtggtagagc cacaaacagc cggtacaagc
       61 aacgatctcc aggaccatct gaatcatgcg cggatgacac gaactcacga cggcgatcac
      121 agacattaac ccacagtaca gacactgcga caacgtggca attcgtcgca atacaacgtg
      181 agaccgaaag tgaaacgtga tttcatgcgt cattttgaac attttgtaaa tcttatttaa
      241 taatgtgtgc ggcaattcac atttaattta tgaatgtttt cttaacatcg cggcaactca
      301 agaaacggca ggttcggatc ttagctacta gagaaagagg agaaatacta gatgcgtaaa
      361 ggcgaagagc tgttcactgg tgtcgtccct attctggtgg aactggatgg tgatgtcaac
      421 ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg caactaatgg taaactgacg
      481 ctgaagttca tctgtactac tggtaaactg ccggttcctt ggccgactct ggtaacgacg
      541 ctgacttatg gtgttcagtg ctttgctcgt tatccggacc atatgaagca gcatgacttc
      601 ttcaagtccg ccatgccgga aggctatgtg caggaacgca cgatttcctt taaggatgac
      661 ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt aaaccgcatt
      721 gagctgaaag gcattgactt taaagaggac ggcaatatcc tgggccataa gctggaatac
      781 aattttaaca gccacaatgt ttacatcacc gccgataaac aaaaaaatgg cattaaagcg
      841 aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc agctggctga tcactaccag
      901 caaaacactc caatcggtga tggtcctgtt ctgctgccag acaatcacta tctgagcacg
      961 caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc atatggttct gctggagttc
     1021 gtaaccgcag cgggcatcac gcatggtatg gatgaactgt acaaatgacc aggcatcaaa
     1081 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     1141 acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttataggt
     1201 ctcagctgga aatctgctcg tcagtggtgc tcacactgac gaatcatgta cagatcatac
     1261 cgatgactgc ctggcgactc acaactaagc aagacagccg gaaccagcgc cggcgaacac
     1321 cactgcatat atggcatatc acaacagtcc acgtctcaag cagttacaga gatgttacga
     1381 accactagtg cactgcagta caaacacagt cctttcccgc aattttcttt ttctattact
     1441 cttggcctcc tctagtacac tctatatttt tttatgcctc ggtaatgatt ttcatttttt
     1501 tttttccacc tagcggatga ctcttttttt ttcttagcga ttggcattat cacataatga
     1561 attatacatt atataaagta atgtgatttc ttcgaagaat atactaaaaa atgagcaggc
     1621 aagataaacg aaggcaaaga tgacagagca gaaagcccta gtaaagcgta ttacaaatga
     1681 aaccaagatt cagattgcga tctctttaaa gggtggtccc ctagcgatag agcactcgat
     1741 cttcccagaa aaagaggcag aagcagtagc agaacaggcc acacaatcgc aagtgattaa
     1801 cgtccacaca ggtatagggt ttctggacca tatgatacat gctctggcca agcattccgg
     1861 ctggtcgcta atcgttgagt gcattggtga cttacacata gacgaccatc acaccactga
     1921 ggactgcggg attgctctcg gtcaagcttt taaagaggcc ctaggggccg tgcgtggagt
     1981 aaaaaggttt ggatcaggat ttgcgccttt ggatgaggca ctttccagag cggtggttga
     2041 tctttcgaac aggccgtacg cagttgtcga acttggtttg caaagggaga aagtaggtga
     2101 tctctcttgc gagatgatcc cgcattttct tgaaagcttt gcagaggcta gcagaattac
     2161 cctccacgtt gattgtctgc gaggcaagaa tgatcatcac cgtagtgaga gtgcgttcaa
     2221 ggctcttgcg gttgccataa gagaagccac ctcgcccaat ggtaccaacg atgttccctc
     2281 caccaaaggt gttcttatgt agtgacaccg attatttaaa gctgctgcat acgatatata
     2341 tacatgtgta tatatgtata cctatgaatg tcagtaagta tgtatacgaa cagtatgata
     2401 ctgaagatga caaggtaatg catcattcta tacgtgtcat tctgaacgag gcgcgctttc
     2461 cttttttctt tttgcttttt cttttttttt ctcttgaact cgacggatca tagagtttct
     2521 acgcaagact tcttgaagtg aaaaccaact ttcagtcttc aaactaaaaa tgaaaatcag
     2581 tgggctaaat tcagcataat acctgtggat ataattacat acaaaaatat aattaaaaat
     2641 atacgttaaa aataatttat ttttgtaaaa actataaaat atattttact ttcggaacaa
     2701 ctttttaact tataattttg ttttaaataa aaacattgta tttaaaaata ataaattatt
     2761 aaataaaatt ttaaagtatg tttatttaaa atatagaata ctattaaaat tactttaaac
     2821 tttaaaataa aaaaataata aattatatag ttttaaaatt aataaatttt atacacgtga
     2881 ccagatcata atagttttct ttccttgccg agcggccgcg tgttacaacc aattaaccaa
     2941 ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     3001 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     3061 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     3121 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     3181 gacgactgaa tccggtgaga atggcaaaag cttatgcatt tctttccaga cttgttcaac
     3241 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     3301 tgattgcgcc tgagcgaggc gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     3361 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     3421 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     3481 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     3541 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     3601 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     3661 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     3721 tcgcggcctg gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     3781 gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta
     3841 acatcagaga ttttgagaca caacgtggct ttgttgaata aatcgaactt ttgctgagtt
     3901 gaaggatcag tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3961 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4021 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4081 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4141 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4201 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4261 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4321 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4381 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4441 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4501 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4561 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4621 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtgcggcc
     4681 gc
//
