LOCUS       sequence-317068-        4681 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 4681)
  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 4681)
  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..4681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             383..394
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             401..1114
                     /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      1126..1197
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1213..1240
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1501..1688
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             1689..2351
                     /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(2996..3811)
                     /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(2996..3811)
                     /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     3719..3738
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3803..3822
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4036..4624
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4525..4544
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgttctttc ctgcgttatc ccctgattct gtggataacc gtgcggccgc ccctgaattc
       61 gcatctagac tgatgagacg tggtagagcc acaaacagcc ggtacaagca acgatctcca
      121 ggaccatctg aatcatgcgc ggatgacacg aactcacgac ggcgatcaca gacattaacc
      181 cacagtacag acactgcgac aacgtggcaa ttcgtcgcaa tacaacgtga gaccgaaagt
      241 gaaacgtgat ttcatgcgtc attttgaaca ttttgtaaat cttatttaat aatgtgtgcg
      301 gcaattcaca tttaatttat gaatgttttc ttaacatcgc ggcaactcaa gaaacggcag
      361 gttcggatct tagctactag agaaagagga gaaatactag atgcgtaaag gcgaagagct
      421 gttcactggt gtcgtcccta ttctggtgga actggatggt gatgtcaacg gtcataagtt
      481 ttccgtgcgt ggcgagggtg aaggtgacgc aactaatggt aaactgacgc tgaagttcat
      541 ctgtactact ggtaaactgc cggttccttg gccgactctg gtaacgacgc tgacttatgg
      601 tgttcagtgc tttgctcgtt atccggacca tatgaagcag catgacttct tcaagtccgc
      661 catgccggaa ggctatgtgc aggaacgcac gatttccttt aaggatgacg gcacgtacaa
      721 aacgcgtgcg gaagtgaaat ttgaaggcga taccctggta aaccgcattg agctgaaagg
      781 cattgacttt aaagaggacg gcaatatcct gggccataag ctggaataca attttaacag
      841 ccacaatgtt tacatcaccg ccgataaaca aaaaaatggc attaaagcga attttaaaat
      901 tcgccacaac gtggaggatg gcagcgtgca gctggctgat cactaccagc aaaacactcc
      961 aatcggtgat ggtcctgttc tgctgccaga caatcactat ctgagcacgc aaagcgttct
     1021 gtctaaagat ccgaacgaga aacgcgatca tatggttctg ctggagttcg taaccgcagc
     1081 gggcatcacg catggtatgg atgaactgta caaatgacca ggcatcaaat aaaacgaaag
     1141 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac
     1201 tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataggtc tcagctggaa
     1261 atctgctcgt cagtggtgct cacactgacg aatcatgtac agatcatacc gatgactgcc
     1321 tggcgactca caactaagca agacagccgg aaccagcgcc ggcgaacacc actgcatata
     1381 tggcatatca caacagtcca cgtctcaagc agttacagag atgttacgaa ccactagtgc
     1441 actgcagtac aaacacagtc ctttcccgca attttctttt tctattactc ttggcctcct
     1501 ctagtacact ctatattttt ttatgcctcg gtaatgattt tcattttttt ttttccacct
     1561 agcggatgac tctttttttt tcttagcgat tggcattatc acataatgaa ttatacatta
     1621 tataaagtaa tgtgatttct tcgaagaata tactaaaaaa tgagcaggca agataaacga
     1681 aggcaaagat gacagagcag aaagccctag taaagcgtat tacaaatgaa accaagattc
     1741 agattgcgat ctctttaaag ggtggtcccc tagcgataga gcactcgatc ttcccagaaa
     1801 aagaggcaga agcagtagca gaacaggcca cacaatcgca agtgattaac gtccacacag
     1861 gtatagggtt tctggaccat atgatacatg ctctggccaa gcattccggc tggtcgctaa
     1921 tcgttgagtg cattggtgac ttacacatag acgaccatca caccactgag gactgcggga
     1981 ttgctctcgg tcaagctttt aaagaggccc taggggccgt gcgtggagta aaaaggtttg
     2041 gatcaggatt tgcgcctttg gatgaggcac tttccagagc ggtggttgat ctttcgaaca
     2101 ggccgtacgc agttgtcgaa cttggtttgc aaagggagaa agtaggtgat ctctcttgcg
     2161 agatgatccc gcattttctt gaaagctttg cagaggctag cagaattacc ctccacgttg
     2221 attgtctgcg aggcaagaat gatcatcacc gtagtgagag tgcgttcaag gctcttgcgg
     2281 ttgccataag agaagccacc tcgcccaatg gtaccaacga tgttccctcc accaaaggtg
     2341 ttcttatgta gtgacaccga ttatttaaag ctgctgcata cgatatatat acatgtgtat
     2401 atatgtatac ctatgaatgt cagtaagtat gtatacgaac agtatgatac tgaagatgac
     2461 aaggtaatgc atcattctat acgtgtcatt ctgaacgagg cgcgctttcc ttttttcttt
     2521 ttgctttttc tttttttttc tcttgaactc gacggatcat agagtttcta cgcaagactt
     2581 cttgaagtga aaaccaactt tcagtcttca aactaaaaat gaaaatcagt gggctaaatt
     2641 cagcataata cctgtggata taattacata caaaaatata attaaaaata tacgttaaaa
     2701 ataatttatt tttgtaaaaa ctataaaata tattttactt tcggaacaac tttttaactt
     2761 ataattttgt tttaaataaa aacattgtat ttaaaaataa taaattatta aataaaattt
     2821 taaagtatgt ttatttaaaa tatagaatac tattaaaatt actttaaact ttaaaataaa
     2881 aaaataataa attatatagt tttaaaatta ataaatttta tacacgtgac cagatcataa
     2941 tagttttctt tccttgccga gcggccgcgt gttacaacca attaaccaat tctgattaga
     3001 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     3061 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     3121 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     3181 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     3241 ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat
     3301 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     3361 gagcgaggcg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     3421 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     3481 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     3541 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     3601 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     3661 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     3721 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctgg
     3781 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     3841 cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat
     3901 tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcagt
     3961 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4021 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4081 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4141 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4201 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4261 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4321 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4381 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4441 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4501 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4561 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4621 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4681 c
//
