LOCUS       sequence-107507-        8210 bp    DNA     circular SYN 14-APR-2025
DEFINITION  harbours marker gene LYS2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8210)
  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 8210)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pK
FEATURES             Location/Qualifiers
     source          1..8210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        165..404
                     /gene="S. cerevisiae LYS2"
                     /label=LYS2 promoter
     CDS             405..4583
                     /codon_start=1
                     /gene="S. cerevisiae LYS2"
                     /product="alpha-aminoadipate reductase, required for lysine
                     biosynthesis"
                     /label=LYS2
                     /note="yeast auxotrophic marker"
                     /translation="MTNEKVWIEKLDNPTLSVLPHDFLRPQQEPYTKQATYSLQLPQLD
                     VPHDSFSNKYAVALSVWAALIYRVTGDDDIVLYIANNKILRFNIQPTWSFNELYSTINN
                     ELNKLNSIEANFSFDELAEKIQSCQDLERTPQLFRLAFLENQDFKLDEFKHHLVDFALN
                     LDTSNNAHVLNLIYNSLLYSNERVTIVADQFTQYLTAALSDPSNCITKISLITASSKDS
                     LPDPTKNLGWCDFVGCIHDIFQDNAEAFPERTCVVETPTLNSDKSRSFTYRDINRTSNI
                     VAHYLIKTGIKRGDVVMIYSSRGVDLMVCVMGVLKAGATFSVIDPAYPPARQTIYLGVA
                     KPRGLIVIRAAGQLDQLVEDYINDELEIVSRINSIAIQENGTIEGGKLDNGEDVLAPYD
                     HYKDTRTGVVVGPDSNPTLSFTSGSEGIPKGVLGRHFSLAYYFNWMSKRFNLTENDKFT
                     MLSGIAHDPIQRDMFTPLFLGAQLYVPTQDDIGTPGRLAEWMSKYGCTVTHLTPAMGQL
                     LTAQATTPFPKLHHAFFVGDILTKRDCLRLQTLAENCRIVNMYGTTETQRAVSYFEVKS
                     KNDDPNFLKKLKDVMPAGKGMLNVQLLVVNRNDRTQICGIGEIGEIYVRAGGLAEGYRG
                     LPELNKEKFVNNWFVEKDHWNYLDKDNGEPWRQFWLGPRDRLYRTGDLGRYLPNGDCEC
                     CGRADDQVKIRGFRIELGEIDTHISQHPLVRENITLVRKNADNEPTLITFMVPRFDKPD
                     DLSKFQSDVPKEVETDPIVKGLIGYHLLSKDIRTFLKKRLASYAMPSLIVVMDKLPLNP
                     NGKVDKPKLQFPTPKQLNLVAENTVSETDDSQFTNVEREVRDLWLSILPTKPASVSPDD
                     SFFDLGGHSILATKMIFTLKKKLQVDLPLGTIFKYPTIKAFAAEIDRIKSSGGSSQGEV
                     VENVTANYAEDAKKLVETLPSSYPSREYFVEPNSAEGKTTINVFVTGVTGFLGSYILAD
                     LLGRSPKNYSFKVFAHVRAKDEEAAFARLQKAGITYGTWNEKFASNIKVVLGDLSKSQF
                     GLSDEKWMDLANTVDIIIHNGALVHWVYPYAKLRDPNVISTINVMSLAAVGKPKFFDFV
                     SSTSTLDTEYYFNLSDKLVSEGKPGILESDDLMNSASGLTGGYGQSKWAAEYIIRRAGE
                     RGLRGCIVRPGYVTGASANGSSNTDDFLLRFLKGSVQLGKIPDIENSVNMVPVDHVARV
                     VVATSLNPPKENELAVAQVTGHPRILFKDYLYTLHDYGYDVEIESYSKWKKSLEASVID
                     RNEENALYPLLHMVLDNLPESTKAPELDDRNAVASLKKDTAWTGVDWSNGIGVTPEEVG
                     IYIAFLNKVGFLPPPTHNDKLPLPSIELTQAQISLVASGAGARGSSAAA"
     rep_origin      complement(4671..5126)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4808..4829)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5020..5039
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5252..5274
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5266..5283
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5267..5283
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5290..5309
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5290..5308
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    5317..5424
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(5436..5454)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5475..5491)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5475..5491)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5488..5510)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5499..5515
                     /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(5523..5529,5530..5547,5548..5553))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5568..5589
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5706..5723)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5877..6465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5957..5976)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6636..7427,7428..7496))
                     /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     7259..7278
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7497..7601)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8183..8201
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcggcgc gtcgaggaaa actcttcaat agttttgcca gcggaattcc acttgcaatt
      181 acataaaaaa ttccggcggt ttttcgcgtg tgactcaatg tcgaaatacc tgcctaatga
      241 acatgaacat cgcccaaatg tatttgaaga cccgctggga gaagttcaag atatataagt
      301 aacaagcagc caatagtata aaaaaaaatc tgagtttatt acctttcctg gaatttcagt
      361 gaaaaactgc taattataga gagatatcac agagttactc actaatgact aacgaaaagg
      421 tctggataga gaagttggat aatccaactc tttcagtgtt accacatgac tttttacgcc
      481 cacaacaaga accttatacg aaacaagcta catattcgtt acagctacct cagctcgatg
      541 tgcctcatga tagtttttct aacaaatacg ctgtcgcttt gagtgtatgg gctgcattga
      601 tatatagagt aaccggtgac gatgatattg ttctttatat tgcgaataac aaaatcttaa
      661 gattcaatat tcaaccaacg tggtcattta atgagctgta ttctacaatt aacaatgagt
      721 tgaacaagct caattctatt gaggccaatt tttcctttga cgagctagct gaaaaaattc
      781 aaagttgcca agatctggaa aggacccctc agttgttccg tttggccttt ttggaaaacc
      841 aagatttcaa attagacgag ttcaagcatc atttagtgga ctttgctttg aatttggata
      901 ccagtaataa tgcgcatgtt ttgaacttaa tttataacag cttactgtat tcgaatgaaa
      961 gagtaaccat tgttgcggac caatttactc aatatttgac tgctgcgcta agcgatccat
     1021 ccaattgcat aactaaaatc tctctgatca ccgcatcatc caaggatagt ttacctgatc
     1081 caactaagaa cttgggctgg tgcgatttcg tggggtgtat tcacgacatt ttccaggaca
     1141 atgctgaagc cttcccagag agaacctgtg ttgtggagac tccaacacta aattccgaca
     1201 agtcccgttc tttcacttat cgcgacatca accgcacttc taacatagtt gcccattatt
     1261 tgattaaaac aggtatcaaa agaggtgatg tagtgatgat ctattcttct aggggtgtgg
     1321 atttgatggt atgtgtgatg ggtgtcttga aagccggcgc aaccttttca gttatcgacc
     1381 ctgcatatcc cccagccaga caaaccattt acttaggtgt tgctaaacca cgtgggttga
     1441 ttgttattag agctgctgga caattggatc aactagtaga agattacatc aatgatgaat
     1501 tggagattgt ttcaagaatc aattccatcg ctattcaaga aaatggtacc attgaaggtg
     1561 gcaaattgga caatggcgag gatgttttgg ctccatatga tcactacaaa gacaccagaa
     1621 caggtgttgt agttggacca gattccaacc caaccctatc tttcacatct ggttccgaag
     1681 gtattcctaa gggtgttctt ggtagacatt tttccttggc ttattatttc aattggatgt
     1741 ccaaaaggtt caacttaaca gaaaatgata aattcacaat gctgagcggt attgcacatg
     1801 atccaattca aagagatatg tttacaccat tatttttagg tgcccaattg tatgtcccta
     1861 ctcaagatga tattggtaca ccgggccgtt tagcggaatg gatgagtaag tatggttgca
     1921 cagttaccca tttaacacct gccatgggtc aattacttac tgcccaagct actacaccat
     1981 tccctaagtt acatcatgcg ttctttgtgg gtgacatttt aacaaaacgt gattgtctga
     2041 ggttacaaac cttggcagaa aattgccgta ttgttaatat gtacggtacc actgaaacac
     2101 agcgtgcagt ttcttatttc gaagttaaat caaaaaatga cgatccaaac tttttgaaaa
     2161 aattgaaaga tgtcatgcct gctggtaaag gtatgttgaa cgttcagcta ctagttgtta
     2221 acaggaacga tcgtactcaa atatgtggta ttggcgaaat aggtgagatt tatgttcgtg
     2281 caggtggttt ggccgaaggt tatagaggat taccagaatt gaataaagaa aaatttgtga
     2341 acaactggtt tgttgaaaaa gatcactgga attatttgga taaggataat ggtgaacctt
     2401 ggagacaatt ctggttaggt ccaagagata gattgtacag aacgggtgat ttaggtcgtt
     2461 atctaccaaa cggtgactgt gaatgttgcg gtagggctga tgatcaagtt aaaattcgtg
     2521 ggttcagaat cgaattagga gaaatagata cgcacatttc ccaacatcca ttggtaagag
     2581 aaaacattac tttagttcgc aaaaatgccg acaatgagcc aacattgatc acatttatgg
     2641 tcccaagatt tgacaagcca gatgacttgt ctaagttcca aagtgatgtt ccaaaggagg
     2701 ttgaaactga ccctatagtt aagggcttaa tcggttacca tcttttatcc aaggacatca
     2761 ggactttctt aaagaaaaga ttggctagct atgctatgcc ttccttgatt gtggttatgg
     2821 ataaactacc attgaatcca aatggtaaag ttgataagcc taaacttcaa ttcccaactc
     2881 ccaagcaatt aaatttggta gctgaaaata cagtttctga aactgacgac tctcagttta
     2941 ccaatgttga gcgcgaggtt agagacttat ggttaagtat attacctacc aagccagcat
     3001 ctgtatcacc agatgattcg tttttcgatt taggtggtca ttctatcttg gctaccaaaa
     3061 tgatttttac cttaaagaaa aagctgcaag ttgatttacc attgggcaca attttcaagt
     3121 atccaacgat aaaggccttt gccgcggaaa ttgacagaat taaatcatcg ggtggatcat
     3181 ctcaaggtga ggtcgtcgaa aatgtcactg caaattatgc ggaagacgcc aagaaattgg
     3241 ttgagacgct accaagttcg tacccctctc gagaatattt tgttgaacct aatagtgccg
     3301 aaggaaaaac aacaattaat gtgtttgtta ccggtgtcac aggatttctg ggctcctaca
     3361 tccttgcaga tttgttagga cgttctccaa agaactacag tttcaaagtg tttgcccacg
     3421 tcagggccaa ggatgaagaa gctgcatttg caagattaca aaaggcaggt atcacctatg
     3481 gtacttggaa cgaaaaattt gcctcaaata ttaaagttgt attaggcgat ttatctaaaa
     3541 gccaatttgg tctttcagat gagaagtgga tggatttggc aaacacagtt gatataatta
     3601 tccataatgg tgcgttagtt cactgggttt atccatatgc caaattgagg gatccaaatg
     3661 ttatttcaac tatcaatgtt atgagcttag ccgccgtcgg caagccaaag ttctttgact
     3721 ttgtttcctc cacttctact cttgacactg aatactactt taatttgtca gataaacttg
     3781 ttagcgaagg gaagccaggc attttagaat cagacgattt aatgaactct gcaagcgggc
     3841 tcactggtgg atatggtcag tccaaatggg ctgctgagta catcattaga cgtgcaggtg
     3901 aaaggggcct acgtgggtgt attgtcagac caggttacgt aacaggtgcc tctgccaatg
     3961 gttcttcaaa cacagatgat ttcttattga gatttttgaa aggttcagtc caattaggta
     4021 agattccaga tatcgaaaat tccgtgaata tggttccagt agatcatgtt gctcgtgttg
     4081 ttgttgctac gtctttgaat cctcccaaag aaaatgaatt ggccgttgct caagtaacgg
     4141 gtcacccaag aatattattc aaagactact tgtatacttt acacgattat ggttacgatg
     4201 tcgaaatcga aagctattct aaatggaaga aatcattgga ggcgtctgtt attgacagga
     4261 atgaagaaaa tgcgttgtat cctttgctac acatggtctt agacaactta cctgaaagta
     4321 ccaaagctcc ggaactagac gataggaacg ccgtggcatc tttaaagaaa gacaccgcat
     4381 ggacaggtgt tgattggtct aatggaatag gtgttactcc agaagaggtt ggtatatata
     4441 ttgcattttt aaacaaggtt ggatttttac ctccaccaac tcataatgac aaacttccac
     4501 tgccaagtat agaactaact caagcgcaaa taagtctagt tgcttcaggt gctggtgctc
     4561 gtggaagctc cgcagcagct taaggttgag cattacgtat gatatgtgaa acagatccta
     4621 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag
     4681 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     4741 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     4801 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     4861 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     4921 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     4981 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     5041 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     5101 gcttaatgcg ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa
     5161 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     5221 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     5281 agtgaattgt aatacgactc actatagggc gaattggagc tccaccgcgg tggcggccgc
     5341 tctagaacta gtgggtcccc cgggctgcag gaattcgata tcaagcttat cgataccgtc
     5401 gacctggagg gggggcccgg taccagcttt tgttcccttt agtgagggtt aatttcgagc
     5461 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5521 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5581 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5641 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5701 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5761 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5821 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5881 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5941 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6001 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6061 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6121 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6181 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6241 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6301 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6361 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6421 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6481 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6541 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6601 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6661 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactgcc cgtcgtgtag
     6721 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6781 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6841 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6901 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6961 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7021 cgagttacat gatcccccat gttgtgaaaa aaagcggtta gctccttcgg tcctccgatc
     7081 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7141 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7201 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7261 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7321 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7381 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7441 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7501 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7561 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7621 ccacctatcg cttgcctgta acttacacgc gcctcgtatc ttttaatgat ggaataattt
     7681 gggaatttac tctgtgttta tttattttta tgttttgtat ttggatttta gaaagtaaat
     7741 aaagaaggta gaagagttac ggaatgaaga aaaaaaaata aacaaaggtt taaaaaattt
     7801 caacaaaaag cgtactttac atatatattt attagacaag aaaagcagat taaatagata
     7861 tacattcgat taacgataag taaaatgtaa aatcacagga ttttcgtgtg tggtcttcta
     7921 cacagacaag atgaaacaat tcggcattaa tacctgagag caggaagagc aagataaaag
     7981 gtagtatttg ttggcgatcc ccctagagtc ttttacatct tcggaaaaca aaaactattt
     8041 tttctttaat ttcttttttt actttctatt tttaatttat atatttatat taaaaaattt
     8101 aaattataat tatttttata gcacgtgatg aaaaggaccc ccgtcctaag aaaccattat
     8161 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
