LOCUS       sequence-107490-        7580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  harbours marker genes LEU2 and MET17.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7580)
  AUTHORS   Mulleder M, Campbell K, Matsarskaia O, Eckerstorfer F, Ralser M
  TITLE     Saccharomyces cerevisiae single-copy plasmids for auxotrophy 
            compensation, multiple marker selection, and for designing 
            metabolically cooperating communities.
  JOURNAL   F1000Res. 2016 Sep 20;5:2351. eCollection 2016.
  PUBMED    27830062
REFERENCE   2  (bases 1 to 7580)
  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: "F1000Res.";
            date: "2016-09-20"; volume: "5"; pages: "2351. eCollection 2016"
COMMENT     Sequence Label: pLM
FEATURES             Location/Qualifiers
     source          1..7580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     165..183
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(221..243)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(376..1710)
                     /codon_start=1
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /product="O-acetylserine and O-acetylhomoserine 
                     sulfhydrylase, required for sulfur amino acid synthesis"
                     /label=MET17
                     /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ
                     LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI
                     VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV
                     PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII
                     VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA
                     SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG
                     FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
     promoter        2193..2600
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2601..3695
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     rep_origin      complement(4230..4689)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4367..4388)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4579..4598
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4815..4837
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4829..4846
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4830..4846
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4853..4872
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4853..4871
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    4880..4987
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(4999..5017)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5038..5054)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5038..5054)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5051..5073)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5062..5078
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5086..5092,5093..5110,5111..5116))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5131..5152
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5269..5286)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5440..6028)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5520..5539)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6199..6990,6991..7059))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6822..6841
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7060..7164)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(7194..7580,1..117)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 ttacatcttc ggaaaacaaa aactattttt tctttaattt ctttttttac tttctatttt
       61 taatttatat atttatatta aaaaatttaa attataatta tttttatagc acgtgatgaa
      121 aaggacccta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
      181 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
      241 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
      301 cgtcagcggg tgttggcggt cgagcttgtg agagaaagta ggtttataca taattttaca
      361 actcattacg cacactcatg gtttttggcc agcgaaaaca gtttcaaaag attgctggaa
      421 gtctgcaata atgtcatcaa taaattcgat accaacagag acacgaatta agtccttggt
      481 aacaccagat gccaactttt ctttgtcatt taattgtttg tgggtagtga agtatggagc
      541 aatgactaag gtcttggcat caccaacatt ggccaagtta gaggcaagct ttaaattgtc
      601 aacaacttga gcaccagaaa gtttgaatgg gtcagtttcc ttgtcggcat ttggtaagtc
      661 ttttacaccg aaagataaga caccaccgaa accgttagat agatacttct tagcattttc
      721 atgatgagaa tgagatgcta aaccagggta tgaaacccaa gatacgtatg gggattgttc
      781 taaccatttg gctaacttca atgcattttc accgtgtctt tcagctctca aagataatgt
      841 ttcaacacct tgtagtagca agaaagaggc aaatgggttc atcaatggac ccaaatctct
      901 taatagttca gttctaacat gaacgatgta tgccaagtta ccgtaggctt cattgtagat
      961 agtaccgtga tatccttcgg caggttgaga gaattgaggg aacttttctg ggtagtcctt
     1021 ccatgggaac ttaccagagt caacaataat accaccgata gtagtaccat gaccaccaat
     1081 ccatttggta gcagaatgtg ttacaatatc agcaccgtat ttaattggct gacagaagta
     1141 accaccggca ccaaatgtgt tgtcaacgac aactggaata ccgtgtttgt gagcaattgc
     1201 aacaattttt tcaaaatccg gaacattgta ctttggatta ccaatggttt ccaaataaac
     1261 agccttggtt ctttcatcaa agaccttttc gaattcttct ggattgtcac cttcaacaaa
     1321 tctagcctcg ataccaaatc ttttgaacga gattttgaac tggttataag taccaccgta
     1381 taagtaagaa gtggaaacga tgttgtcacc agtgtgtgcc aaaccttgga tggcaagggt
     1441 ttgagcggct tgaccggagg aaacagccaa agcagcagca ccaccttcta aagcagcaat
     1501 tctttcttcc aaaacattac tggttgggtt ttggaaacgg gaatagacgt aacctggaac
     1561 ttctagacca aacaattgcg aaccatgctt agagttttcg aaaacataag aagtggtggc
     1621 gtaaattggt acagctctgg atctgtgagc attgtcacca gggttctctt ggccggcgtg
     1681 tagttgaaca gtatcgaaat gagatggcat tgtatggatg ggggtaatag aattgtatct
     1741 atgtatctga cgaccctgta ttacacgaag gaaatagtag atgaaaagac aagagagcaa
     1801 gaaaaaggaa aaacttcctt tctaacagac gctttactta accttatata tatcttattt
     1861 ttttcatcga gcgtgttcaa ttggacaagg tgccaccttt tcgacacttc ggttattatg
     1921 ttacacagtt ttcatgagga tggcgcgcgc catcgaggag aacttctagt atatctacat
     1981 acctaatatt attgccttat taaaaatgga atcccaacaa ttacatcaaa atccacattc
     2041 tcttcaaaat caattgtcct gtacttcctt gttcatgtgt gttcaaaaac gttatattta
     2101 taggataatt atactctatt tctcaacaag taattggttg tttggccgag cggtctaagg
     2161 cgcctgattc aagaaatatc ttgaccgcag ttaactgtgg gaatactcag gtatcgtaag
     2221 atgcaagagt tcgaatctct tagcaaccat tatttttttc ctcaacataa cgagaacaca
     2281 caggggcgct atcgcacaga atcaaattcg atgactggaa attttttgtt aatttcagag
     2341 gtcgcctgac gcatatacct ttttcaactg aaaaattggg agaaaaagga aaggtgagag
     2401 cgccggaacc ggcttttcat atagaataga gaagcgttca tgactaaatg cttgcatcac
     2461 aatacttgaa gttgacaata ttatttaagg acctattgtt ttttccaata ggtggttagc
     2521 aatcgtctta ctttctaact tttcttacct tttacatttc agcaatatat atatatatat
     2581 ttcaaggata taccattcta atgtctgccc ctaagaagat cgtcgttttg ccaggtgacc
     2641 acgttggtca agaaatcaca gccgaagcca ttaaggttct taaagctatt tctgatgttc
     2701 gttccaatgt caagttcgat ttcgaaaatc atttaattgg tggtgctgct atcgatgcta
     2761 caggtgttcc acttccagat gaggcgctgg aagcctccaa gaaggctgat gccgttttgt
     2821 taggtgctgt gggtggtcct aaatggggta ccggtagtgt tagacctgaa caaggtttac
     2881 taaaaatccg taaagaactt caattgtacg ccaacttaag accatgtaac tttgcatccg
     2941 actctctttt agacttatct ccaatcaagc cacaatttgc taaaggtact gacttcgttg
     3001 ttgtcagaga attagtggga ggtatttact ttggtaagag aaaggaagac gatggtgatg
     3061 gtgtcgcttg ggatagtgaa caatacaccg ttccagaagt gcaaagaatc acaagaatgg
     3121 ccgctttcat ggccctacaa catgagccac cattgcctat ttggtccttg gataaagcta
     3181 atgttttggc ctcttcaaga ttatggagaa aaactgtgga ggaaaccatc aagaacgaat
     3241 tccctacatt gaaggttcaa catcaattga ttgattctgc cgccatgatc ctagttaaga
     3301 acccaaccca cctaaatggt attataatca ccagcaacat gtttggtgat atcatctccg
     3361 atgaagcctc cgttatccca ggttccttgg gtttgttgcc atctgcgtcc ttggcctctt
     3421 tgccagacaa gaacaccgca tttggtttgt acgaaccatg ccacggttct gctccagatt
     3481 tgccaaagaa taaggtcaac cctatcgcca ctatcttgtc tgctgcaatg atgttgaaat
     3541 tgtcattgaa cttgcctgaa gaaggtaagg ccattgaaga tgcagttaaa aaggttttgg
     3601 atgcaggtat cagaactggt gatttaggtg gttccaacag taccaccgaa gtcggtgatg
     3661 ctgtcgccga agaagttaag aaaatccttg cttaaaaaga ttctcttttt ttatgatatt
     3721 tgtacataaa ctttataaat gaaattcata atagaaacga cacgaaatta caaaatggaa
     3781 tatgttcata gggtagacga aactatatac gcaatctaca tacatttatc aagaaggaga
     3841 aaaaggagga tgtaaaggaa tacaggtaag caaattgata ctaatggctc aacgtgataa
     3901 ggaaaaagaa ttgcacttta acattaatat tgacaaggag gagggcacca cacaaaaagt
     3961 taggtgtaac agaaaatcat gaaactatga ttcctaattt atatattgga ggattttctc
     4021 taaaaaaaaa aaaatacaac aaataaaaaa cactcaatga cctgaccatt tgatggagtt
     4081 taagtcaata ccttcttgaa ccatttccca taatggtgaa agttccctca agaattttac
     4141 tctgtcagaa acggccttac gacgtagtcg aggatcctat gcggtgtgaa ataccgcaca
     4201 gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt
     4261 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     4321 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     4381 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4441 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     4501 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4561 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggctgg
     4621 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     4681 agggcgcgtc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     4741 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     4801 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga
     4861 ctcactatag ggcgaattgg agctccaccg cggtggcggc cgctctagaa ctagtgggtc
     4921 ccccgggctg caggaattcg atatcaagct tatcgatacc gtcgacctgg agggggggcc
     4981 cggtaccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt aatcatggtc
     5041 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5101 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5161 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5221 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5281 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5341 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5401 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5461 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5521 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5581 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5641 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5701 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5761 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5821 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5881 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5941 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6001 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6061 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6121 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6181 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6241 tctatttcgt tcatccatag ttgcctgact gcccgtcgtg tagataacta cgatacggga
     6301 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6361 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6421 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6481 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6541 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6601 catgttgtga aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6661 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6721 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6781 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6841 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6901 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6961 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7021 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7081 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7141 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acggatcgct
     7201 tgcctgtaac ttacacgcgc ctcgtatctt ttaatgatgg aataatttgg gaatttactc
     7261 tgtatttatt tatttttatg ttttgtattt ggattttaga aagtaaataa agaaggtaga
     7321 agagttacgg aatgaagaaa aaaaaataaa caaaggttta aaaaatttca acaaaaagcg
     7381 tactttacat atatatttat tagacaagaa aagcagatta aatagatata cattcgatta
     7441 acgataagta aaatgtaaaa tcacaggatt ttcgtgtgtg gtcttctaca cagacaagat
     7501 gaaacaattc ggcattaata cctgagagca ggaagagcaa gataaaaggt agtatttgtt
     7561 ggcgatcccc ctagagtctt
//
