LOCUS       sequence-108551-        7603 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative E. coli/S. cerevisiae shuttle vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7603)
  AUTHORS   Gnugge R, Liphardt T, Rudolf F
  TITLE     A shuttle vector series for precise genetic engineering of 
            Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2015 Dec 9. doi: 10.1002/yea.3144.
  PUBMED    26647923
REFERENCE   2  (bases 1 to 7603)
  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: "Yeast. 2015
            Dec 9. doi: 10.1002/yea.3144."
COMMENT     Sequence Label: pRG207
FEATURES             Location/Qualifiers
     source          1..7603
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(372..4550)
                     /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"
     promoter        complement(4551..4790)
                     /gene="S. cerevisiae LYS2"
                     /label=LYS2 promoter
     primer_bind     4892..4914
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4906..4923
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4907..4923
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4930..4949
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4930..4948
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     5057..5073
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5057..5073
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5107..5123)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5108..5124)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(5222..5240)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5261..5277)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5261..5277)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5274..5296)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5285..5301
                     /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)."
     primer_bind     complement(5675..5692)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5846..6434)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5926..5945)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6613..7404,7405..7473))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7236..7255
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7474..7578)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgaaa gaacgcggca ttagcaacat ttgcgaagcg aacgattttg cgaactgcta
       61 ttaattaatt ggcgcgcctg tgaagatcac aaggttttgc attgaggata gtatagaagc
      121 aagaatcatt gaattacagg aaaaaaaggc aaatatgatt catgctacaa taaaccaaga
      181 tgaagctgcc attagcagac taacgccagc cgatttacag ttcttattca ataactaata
      241 ttttattctc ttattatata ttattctcgg agtttttaag tgacatcacc cgaaaagaag
      301 ctaagtcttt ctcctaattc atatttaatt attgtacatg gacatatgca tatcatacgt
      361 aatgctcaac cttaagctgc tgcggagctt ccacgagcac cagcacctga agcaactaga
      421 cttatttgcg cttgagttag ttctatactt ggcagtggaa gtttgtcatt atgagttggt
      481 ggaggtaaaa atccaacctt gtttaaaaat gcaatatata taccaacctc ttctggagta
      541 acacctattc cattagacca atcaacacct gtccatgcgg tgtctttctt taaagatgcc
      601 acggcgttcc tatcgtctag ttccggagct ttggtacttt caggtaagtt gtctaagacc
      661 atgtgtagca aaggatacaa cgcattttct tcattcctgt caataacaga cgcctccaat
      721 gatttcttcc atttagaata gctttcgatt tcgacatcgt aaccataatc gtgtaaagta
      781 tacaagtagt ctttgaataa tattcttggg tgacccgtta cttgagcaac ggccaattca
      841 ttttctttgg gaggattcaa agacgtagca acaacaacac gagcaacatg atctactgga
      901 accatattca cggaattttc gatgtctgga atcttaccta attggactga acctttcaaa
      961 aatctcaata agaaatcatc tgtgtttgaa gaaccattgg cagaggcacc tgttacgtaa
     1021 cctggtctga caatacaccc acgtaggccc ctttcacctg cacgtctaat gatgtactca
     1081 gcagcccatt tggactgacc atatccacca gtgagcccgc ttgcagagtt cattaaatcg
     1141 tctgattcta aaatgcctgg cttcccttcg ctaacaagtt tatctgacaa attaaagtag
     1201 tattcagtgt caagagtaga agtggaggaa acaaagtcaa agaactttgg cttgccgacg
     1261 gcggctaagc tcataacatt gatagttgaa ataacatttg gatctctcaa tttggcgtat
     1321 ggataaaccc agtgaactaa cgcaccatta tggataatta tatcaactgt gtttgccaaa
     1381 tccatccact tctcatctga aagaccaaat tggcttttag ataaatcgcc taatacaact
     1441 ttaatatttg aggcaaattt ttcgttccaa gtaccatagg tgatacctgc cttttgtaat
     1501 cttgcaaatg cagcttcttc atccttggcc ctgacgtggg caaacacttt gaaactgtag
     1561 ttctttggag aacgtcctaa caaatctgca aggatgtagg agcccagaaa tcctgtgaca
     1621 ccagtaacaa acacattaat tgttgttttt ccttcggcac tattaggttc aacaaaatat
     1681 tcacgagagg ggtacgaact tggtagcgtc tcaaccaatt tcttggcgtc ttccgcataa
     1741 tttgcagtga cattttcgac gacctcacct tgagatgatc cacccgatga tttaattctg
     1801 tcaatttccg cggcaaaggc ctttatcgtt ggatacttga aaattgtgcc caatggtaaa
     1861 tcaacttgca gctttttctt taaggtaaaa atcattttgg tagccaagat agaatgacca
     1921 cctaaatcga aaaacgaatc atctggtgat acagatgctg gcttggtagg taatatactt
     1981 aaccataagt ctctaacctc gcgctcaaca ttggtaaact gagagtcgtc agtttcagaa
     2041 actgtatttt cagctaccaa atttaattgc ttgggagttg ggaattgaag tttaggctta
     2101 tcaactttac catttggatt caatggtagt ttatccataa ccacaatcaa ggaaggcata
     2161 gcatagctag ccaatctttt ctttaagaaa gtcctgatgt ccttggataa aagatggtaa
     2221 ccgattaagc ccttaactat agggtcagtt tcaacctcct ttggaacatc actttggaac
     2281 ttagacaagt catctggctt gtcaaatctt gggaccataa atgtgatcaa tgttggctca
     2341 ttgtcggcat ttttgcgaac taaagtaatg ttttctctta ccaatggatg ttgggaaatg
     2401 tgcgtatcta tttctcctaa ttcgattctg aacccacgaa ttttaacttg atcatcagcc
     2461 ctaccgcaac attcacagtc accgtttggt agataacgac ctaaatcacc cgttctgtac
     2521 aatctatctc ttggacctaa ccagaattgt ctccaaggtt caccattatc cttatccaaa
     2581 taattccagt gatctttttc aacaaaccag ttgttcacaa atttttcttt attcaattct
     2641 ggtaatcctc tataaccttc ggccaaacca cctgcacgaa cataaatctc acctatttcg
     2701 ccaataccac atatttgagt acggtcgttc ctgttaacaa ctaatagctg aacgttcaac
     2761 atacctttac cagcaggcat gacatctttc aattttttca aaaagtttgg atcgtcattt
     2821 tttgatttaa cttcgaaata agaaactgca cgctgtgttt cagtagtacc gtacatatta
     2881 acaatacggc aattttctgc caaggtttgt aacctcagac aatcacgttt tgttaaaatg
     2941 tcacccacaa agaacgcatg atgtaactta gggaatggtg tagtagcttg ggcagtaagt
     3001 aattgaccca tggcaggtgt taaatgggta actgtgcaac catacttact catccattcc
     3061 gctaaacggc ccggtgtacc aatatcatct tgagtaggga catacaattg ggcacctaaa
     3121 aataatggtg taaacatatc tctttgaatt ggatcatgtg caataccgct cagcattgtg
     3181 aatttatcat tttctgttaa gttgaacctt ttggacatcc aattgaaata ataagccaag
     3241 gaaaaatgtc taccaagaac acccttagga ataccttcgg aaccagatgt gaaagatagg
     3301 gttgggttgg aatctggtcc aactacaaca cctgttctgg tgtctttgta gtgatcgtat
     3361 ggagccaaaa catcctcgcc attgtccaat ttgccacctt caatagtacc attttcttga
     3421 atagcgatgg aattgattct tgaaacaatc tccaattcat cattgatgta atcttctact
     3481 aattgatcca attgtccagc agctctaata acaatcaacc cacgtggttt agcaacacct
     3541 aagtaaatgg tttgtctggc tgggggatat gcagggtcga taactgaaaa ggttgcgcca
     3601 gctttcaaga cacccatcac acataccatc aaatccacac ccctagaaga atagatcatc
     3661 actacatcac ctcttttgat acctgtttta atcaaataat gggcaactat gttagaagtg
     3721 cggttgatgt cgcgataagt gaaagaacgg gacttgtcgg aatttagtgt tggagtctcc
     3781 acaacacagg ttctctctgg gaaggcttca gcattgtcct ggaaaatgtc gtgaatacac
     3841 cccacgaaat cgcaccagcc caagttctta gttggatcag gtaaactatc cttggatgat
     3901 gcggtgatca gagagatttt agttatgcaa ttggatggat cgcttagcgc agcagtcaaa
     3961 tattgagtaa attggtccgc aacaatggtt actctttcat tcgaatacag taagctgtta
     4021 taaattaagt tcaaaacatg cgcattatta ctggtatcca aattcaaagc aaagtccact
     4081 aaatgatgct tgaactcgtc taatttgaaa tcttggtttt ccaaaaaggc caaacggaac
     4141 aactgagggg tcctttccaa gtcttggcaa ctttgaattt tttcagctag ctcgtcaaag
     4201 gaaaaattgg cctcaataga attgagcttg ttcaactcat tgttaattgt agaatacagc
     4261 tcattaaatg accacgttgg ttgaatattg aatcttaaga ttttgttatt cgcaatataa
     4321 agaacaatat catcgtcacc agttactcta tatatcaatg cagcccatac actcaaagcg
     4381 acagcgtatt tgttagaaaa actatcatga ggcacatcga gctgaggtag ctgtaacgaa
     4441 tatgtagctt gtttcgtata aggttcttgt tgtgggcgta aaaagtcatg tggtaacact
     4501 gaaagagttg gattatccaa cttctctatc cagacctttt cgttagtcat tagtgagtaa
     4561 ctctgtgata tgtctctata attagcagtt tttcactgaa attccaggaa aggtaataaa
     4621 ctcagatttt tttttatact attggctgct tgttacttat atatcttgaa cttctcccag
     4681 cgggtcttca aatacatttg ggcgatgttc atgttcatta ggcaggtatt tcgacattga
     4741 gtcacacgcg aaaaaccgcc ggaatttttt atgtaattgc aagtgcaatt ccgctggcaa
     4801 aactattgaa gagttttcct cgctaaaact gtgcgatgcc tctcgatcgg atgtgctgca
     4861 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     4921 agtgaattgt aatacgactc actatagggc gaattggaga aacagaataa tatataaacc
     4981 tgtataatat aaccttgaag actatatttc ttttcgcggc cgcctatagg gcgaattgga
     5041 gctccaccgc ggtggctcta gaactagtgg atcccccggg ctgcaggaat tcgatatcaa
     5101 gcttatcgat accgtcgacc tcgagggggg gcccggtacc cagcttcggc cgtatttaca
     5161 gataacgatg tcgttattat atatatatat atatattgtc acttgtaccc agcttttgtt
     5221 ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5281 gaaattgtta tccgctcaca acagctgaga agcggtcagc aagaagaaaa tcgaataatc
     5341 agtttcaatg aatatagacg taacaaaggc acaaaaaaga aaacaagtag atatgtcata
     5401 catatatata tatttcaggc agcaagtgac catcaaccag gaagtgtgaa atagcaaaga
     5461 actgtttact tcttcagact tagaaagctc ttcccatagg agaaagcaac tgctttcctt
     5521 tgtagtaatt gattgtcttt tcagcccttg acgcaacacc gtccgctgtg aaatcgaaca
     5581 atttgtaaat ttcaggcgcg ccatcctgca ggcattaatg aatcggccaa cgcgcgggga
     5641 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5701 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5761 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5821 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5881 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5941 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6001 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6061 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6121 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6181 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6241 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6301 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6361 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6421 aaaaaggatc tcaagaagat cctttgatct tttgtttaaa cctacggggt ctgacgctca
     6481 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6541 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6601 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6661 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6721 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6781 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6841 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6901 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6961 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7021 gtgcaaaaaa gcggttagct ccttcggtcc tccgatggtt gtcagaagta agttggccgc
     7081 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7141 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7201 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7261 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7321 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7381 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7441 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7501 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7561 acaaataggg gttccgcgca catttccccg aaaagtgcca cct
//
