LOCUS       sequence-234491-        6594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpession plasmid of glycerol kinase (GUT1) in Y. lipolytica.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6594)
  AUTHORS   Egermeier M, Sauer M, Marx H
  TITLE     Golden Gate based metabolic engineering strategy for wild-type 
            strains of Yarrowia lipolytica.
  JOURNAL   FEMS Microbiol Lett. 2019 Jan 29. pii: 5304171. doi: 
            10.1093/femsle/fnz022.
  PUBMED    30698703
REFERENCE   2  (bases 1 to 6594)
  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: "FEMS 
            Microbiol Lett. 2019 Jan 29. pii: 5304171. doi: 
            10.1093/femsle/fnz022."
COMMENT     Sequence Label: pMEG_BB3_YL001
FEATURES             Location/Qualifiers
     source          1..6594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(2396..2984)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2476..2495)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     gene            5475..6594
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        5475..5818
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             5819..6391
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      6397..6594
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 gcttggatcc gaattttcag aacctaattt atctgttacc cggcctgtgg ctcgcacagc
       61 ttaaaaatgg tcaaactttc cccttcttgt ctttttttcc tcacattcat caggttcttg
      121 tcttgatctt tcaagtgagt attaattacc gaccttggtt cttcattggg agagcattgg
      181 aagccgtggt gcagcaacca caaaacggtt cttccccttc gataccttct tgcctgcctt
      241 tcaatacaag tcggctcgat tagcggtggt cgcccccgcc agcggagaac atggaactaa
      301 cccagaatga gagctaagtg gagaaagaag agagtcagac gactcaagcg aaagcgccgc
      361 aaggtccgag ctcgatccaa ataagcggtt tttaacggag atttaacact aaatcgaaga
      421 acttttcccg tttcatttgc gaatgagctc gttaacaaaa tcccccagtt tttttatcca
      481 gctgtaagga ttgacattag taatgaatta ttgtttggta tatttaaatc tgtagttcct
      541 ttctgtccgt gtcggcaact gttgtactcg tgatttactt gtattgacga atacttactg
      601 tagcgcactc tgctgctact ggtcgtaagg atgtgctatt tcggtgtatg gtgggttttt
      661 tgggggtcgg aaccgatgac tgttacacgg gcacggctcg ttgtgtacac gcacagagct
      721 cttgcgagtc atgttgtagc tagctcgtcg tgttcaggaa ctgttcgatg gttcggagag
      781 agtcgccgcc cagaacatac gcgcaccgat gtcagcagac agccttatta caagtacagt
      841 atgtacatac tactgtatat tcaagcaagt atatccgtgg gtgcgggtga tttggatcta
      901 aggttcgtac tcaacactca cgagcagctt gcctatgtta catcctttta tcagacataa
      961 cataattgga gtttacttac acacggggtg tacctgtatg agcaccacct acaattgtag
     1021 cactggtact tgtacaaaga atttattcgt acgaatcaca ggtacggccg ccctcaccga
     1081 accagcgaat acctcagcgg tcccctgcag tgactcaaca aagcgatatg aacatcttgc
     1141 gatggtatcc tgctgatagt ttttactgta caaacacctg tgtagctcct tctagcattt
     1201 ttaagttatt cacacctcaa ggcgagggat aaattaaata aattccaaaa gcgaagatcg
     1261 agaaactaaa ttaaaattcc aaaaacgaag ttggaacaca accccccgaa aaaaaacaac
     1321 aaacaaaaaa cccaacaaaa taaacaaaaa caaaataaat atataactac cagtatctga
     1381 ctaaaagttc aaatactcgt acttacaaca aatagaaatg agccggccaa aattctgcag
     1441 aaaaaaattt caaacaagta ctggtataat taaattaaaa aacacatcaa agtatcataa
     1501 cgttagttat tttattttat ttaataaaag aaaacaacaa aatgggctca aaactttcaa
     1561 cttatacgat acataccaaa taacaattta gtatttatct aagtgctttt cgtagataat
     1621 ggaatacaaa tggatatcca gagtatacac atggatagta tacactgaca cgacaattct
     1681 gtatctcttt atgttaacta ctgtgaggcg ttaaatagag cttgatatat aaaatgttac
     1741 atttcacagt ctgaactttt gcagattacc taatttggta agatattaat tatgaactga
     1801 aagttgatgg catccctaaa tttgatgaaa gatgaaattg taaatgaggt ggtaaaagag
     1861 ctacagtcgt tttgttttga gataccatca tctctaacga aatatctatt aaaaatctca
     1921 gtgtgatcat gagtcattgc caccctggaa aatgtcatca tggctgatat ttctaactgt
     1981 ttacttgaga taaatatata tttacaagaa cttcccttga aattaattta gatataaaat
     2041 gtttgcgggc aagttactac gaggaataaa ttatatctgt tgactagaag ttatgaacat
     2101 tcagtatata tgcacatata ataaccaact tcgaattttc caaattagga agtaaattgt
     2161 tataatgaaa catactttta ggtttttata tattataatg gaatacgtct ccattatttt
     2221 cgccatctaa tttatttaat aaaaaagttt agatagatga tctaatttta tatattaaaa
     2281 tagatggtga gagaaaaaaa aatggcttgc atatatatat atatatacag atcccgctgt
     2341 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2401 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2461 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2521 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2581 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2641 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2701 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2761 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2821 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2881 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2941 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3001 tttctacggg gtggagcgca aaaaaccccg cccctgacag ggcggggttt tttcgcgatc
     3061 ggagagacac cgggttggcg gcgcatttgt gtcccaaaaa acagccccaa ttgccccaat
     3121 tgaccccaaa ttgacccagt agcgggccca accccggcga gagccccctt ctccccacat
     3181 atcaaacctc ccccggttcc cacacttgcc gttaagggcg tagggtactg cagtctggaa
     3241 tctacgcttg ttcagacttt gttctagttt ctttgtctgg ccatccgggt aacccatgcc
     3301 ggacgcaaaa tagactactg aaaatttttt tgctttgtgg ttgggacttt agccaagggt
     3361 ataaaagacc accgtccccg aattaccttt cctcttcttt tctctctctc cttgtcaact
     3421 cacacccgaa atcgttaagc atttccttct gagtataaga atcattcaac atgtcatcct
     3481 acgtaggagc tctcgaccag ggtaccacct ccacccgttt cattctcttt tcgcctgacg
     3541 gcaagcccgt ggcatcccac cagatcgaat tcacccagat ctacccccac cccggatggg
     3601 tggagcacga ccccgaggag ctcgtgagct cgtgtctgga gtgcatgtcg tcggtggcca
     3661 aggaaatgcg aacccagggc atcaaggtgg ccgacgtgaa ggcgatcgga atcaccaacc
     3721 agcgagaaac caccgtgctt tgggacattg agacgggcca gcccctgtac aacgccattg
     3781 tgtggtccga cgcccgaacc ggcgacaccg tcaagaagct cgaggcccag cccggcgctg
     3841 acgaaatccc caagctctgt ggcctgcccc tgtccaccta ctttgccgga gtcaaggtcc
     3901 gatggatcct ggataacgtc aaggaggccc gagagtgcta cgatcgaggc aagctggcct
     3961 tctccaccat cgactcgtgg ctgctctaca acctcacggg cggcctcaac ggcggcgccc
     4021 acatcaccga cacctccaac gcctcccggt ccatgttcat gaacattgag acgctcaagt
     4081 acgacgagaa gctcatcaag ttctttggcg tcgagaagct cattctcccc aagattgtct
     4141 cgtccgccga ggtctacggc cgaatcggaa ccggcccctt cgccaacatc cccctggccg
     4201 gctgtctcgg tgaccagtcc gccgccctcg tcggccagaa ggcctttgag cccggccagg
     4261 ccaagaacac atatggaacc ggctgcttcc tgctctacaa cgccggcgag aagcccatca
     4321 tctccaacaa cggcctgctg accaccgtcg gctaccactt caagggccag aagcccgtct
     4381 acgctctgga gggctccatc tccgtcgccg gctcgtgcat caagtggctg cgagacaaca
     4441 ttggtctgat tgagtcatcc gagcagattg gagagcttgc ctcccaggtc gacgactctg
     4501 ccggcgtggt gtttgtcacc gctctgtccg gcctgtttgc cccctactgg cgaaccgacg
     4561 cccgaggcac cattctgggt ctgactcagt tcaccaccaa ggcccacatt tgccgagccg
     4621 ccctggaggc tacttgtttc cagacccggg ccattctcga cgccatggcc aaggactctg
     4681 gtaagccctt caccaagctg cgagtcgacg gaggaatgac caactcggac attgctatgc
     4741 agatccaggc cgacattctt ggcattgagg tcgagcgacc cgccatgcga gagacgaccg
     4801 ctctgggtgc cgccattgct gccggctttg ccgttggcgt gtggaagtcc attgaggatc
     4861 ttaaggacat caacaccgag ggcatgaccg agtttgcttc caagaccaac gaggaggagc
     4921 gggccgccat gatgaagcag tggaaccggg gcattgagcg agctgttggc tggcttgagt
     4981 aagcttgcgt ctacaactgg acccttagcc tgtatatatc aattgattat ttaaagattt
     5041 ggtcggtagg cggttcgtat tgtacaatgg gatctgttac tgaggtggat ctacccaact
     5101 tgcgagattc aattgcgaga ttcaatcgcg agattcaatt gcgagaatca gttgcgagtt
     5161 gttctaacac tcagcttcta cgagcgcttg tattaggacg agtgatactc cgtggggcga
     5221 cggcttctct tgcgtcttgt gttgtattct ttcttacact atcgtccatc tccaaccacc
     5281 tcgtactcgc acgctcgcct gtactgctta ctccaaacaa catgactggc tgacgaaatc
     5341 aaggcgctgt atatatgtac acatctacag caacagaagg ttgttttgtt gcccagctcg
     5401 ctaatagtct tgaaaccgtc tccattcaca gctgagacat gtggaggttg cggttcgatt
     5461 gacgctccgg catggacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca
     5521 gctcaggggc atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat
     5581 catttgcatc catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc
     5641 tgcagacctg cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg
     5701 cgcccctgta gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac
     5761 ttccttttaa aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaccat
     5821 gggtaccact cttgacgaca cggcttaccg gtaccgcacc agtgtcccgg gggacgccga
     5881 ggccatcgag gcactggatg ggtccttcac caccgacacc gtttttcgcg tcaccgccac
     5941 cggggacggc ttcaccctgc gggaggtgcc ggtggacccg cccctgacca aggtgttccc
     6001 cgacgacgaa tcggacgacg aatcggacga cggggaggac ggcgacccgg actcccggac
     6061 gttcgtcgcg tacggggacg acggcgacct ggcgggcttc gtggtcgtct cgtactccgg
     6121 ctggaaccgc cggctgaccg tcgaggacat cgaggtcgcc ccggagcacc gggggcacgg
     6181 ggtcgggcgc gcgttgatgg ggctcgcgac ggagttcgcc cgcgagcggg gcgccgggca
     6241 cctctggctg gaggtcacca acgtcaacgc accggcgatc cacgcgtacc ggcggatggg
     6301 gttcaccctc tgcggcctgg acaccgccct gtacgacggc accgcctcgg acggcgagca
     6361 ggcgctctac atgagcatgc cctgccccta atcagtactg acaataaaaa gattcttgtt
     6421 ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat
     6481 gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt
     6541 gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actg
//
