LOCUS       sequence-108552-        8068 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Centromeric E. coli/S. cerevisiae shuttle vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8068)
  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 8068)
  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: pRG217
FEATURES             Location/Qualifiers
     source          1..8068
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    18..521
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(837..5015)
                     /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(5016..5255)
                     /gene="S. cerevisiae LYS2"
                     /label=LYS2 promoter
     primer_bind     5357..5379
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5371..5388
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5372..5388
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5395..5414
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5395..5413
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     5522..5538
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5522..5538
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5572..5588)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5573..5589)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(5687..5705)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5726..5742)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5726..5742)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5739..5761)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5750..5766
                     /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(6140..6157)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6311..6899)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6391..6410)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7078..7869,7870..7938))
                     /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     7701..7720
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7939..8043)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgggt ccttttcatc acgtgctata aaaataatta taatttaaat tttttaatat
       61 aaatatataa attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt
      121 ttccgaagat gtaaaagact ctagggggat cgccaacaaa tactaccttt tatcttgctc
      181 ttcctgctct caggtattaa tgccgaattg tttcatcttg tctgtgtaga agaccacaca
      241 cgaaaatcct gtgattttac attttactta tcgttaatcg aatgtatatc tatttaatct
      301 gcttttcttg tctaataaat atatatgtaa agtacgcttt ttgttgaaat tttttaaacc
      361 tttgtttatt tttttttctt cattccgtaa ctcttctacc ttctttattt actttctaaa
      421 atccaaatac aaaacataaa aataaataaa cacagagtaa attcccaaat tattccatca
      481 ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat ccgtcttaat taattggcgc
      541 gcctgtgaag atcacaaggt tttgcattga ggatagtata gaagcaagaa tcattgaatt
      601 acaggaaaaa aaggcaaata tgattcatgc tacaataaac caagatgaag ctgccattag
      661 cagactaacg ccagccgatt tacagttctt attcaataac taatatttta ttctcttatt
      721 atatattatt ctcggagttt ttaagtgaca tcacccgaaa agaagctaag tctttctcct
      781 aattcatatt taattattgt acatggacat atgcatatca tacgtaatgc tcaaccttaa
      841 gctgctgcgg agcttccacg agcaccagca cctgaagcaa ctagacttat ttgcgcttga
      901 gttagttcta tacttggcag tggaagtttg tcattatgag ttggtggagg taaaaatcca
      961 accttgttta aaaatgcaat atatatacca acctcttctg gagtaacacc tattccatta
     1021 gaccaatcaa cacctgtcca tgcggtgtct ttctttaaag atgccacggc gttcctatcg
     1081 tctagttccg gagctttggt actttcaggt aagttgtcta agaccatgtg tagcaaagga
     1141 tacaacgcat tttcttcatt cctgtcaata acagacgcct ccaatgattt cttccattta
     1201 gaatagcttt cgatttcgac atcgtaacca taatcgtgta aagtatacaa gtagtctttg
     1261 aataatattc ttgggtgacc cgttacttga gcaacggcca attcattttc tttgggagga
     1321 ttcaaagacg tagcaacaac aacacgagca acatgatcta ctggaaccat attcacggaa
     1381 ttttcgatgt ctggaatctt acctaattgg actgaacctt tcaaaaatct caataagaaa
     1441 tcatctgtgt ttgaagaacc attggcagag gcacctgtta cgtaacctgg tctgacaata
     1501 cacccacgta ggcccctttc acctgcacgt ctaatgatgt actcagcagc ccatttggac
     1561 tgaccatatc caccagtgag cccgcttgca gagttcatta aatcgtctga ttctaaaatg
     1621 cctggcttcc cttcgctaac aagtttatct gacaaattaa agtagtattc agtgtcaaga
     1681 gtagaagtgg aggaaacaaa gtcaaagaac tttggcttgc cgacggcggc taagctcata
     1741 acattgatag ttgaaataac atttggatct ctcaatttgg cgtatggata aacccagtga
     1801 actaacgcac cattatggat aattatatca actgtgtttg ccaaatccat ccacttctca
     1861 tctgaaagac caaattggct tttagataaa tcgcctaata caactttaat atttgaggca
     1921 aatttttcgt tccaagtacc ataggtgata cctgcctttt gtaatcttgc aaatgcagct
     1981 tcttcatcct tggccctgac gtgggcaaac actttgaaac tgtagttctt tggagaacgt
     2041 cctaacaaat ctgcaaggat gtaggagccc agaaatcctg tgacaccagt aacaaacaca
     2101 ttaattgttg tttttccttc ggcactatta ggttcaacaa aatattcacg agaggggtac
     2161 gaacttggta gcgtctcaac caatttcttg gcgtcttccg cataatttgc agtgacattt
     2221 tcgacgacct caccttgaga tgatccaccc gatgatttaa ttctgtcaat ttccgcggca
     2281 aaggccttta tcgttggata cttgaaaatt gtgcccaatg gtaaatcaac ttgcagcttt
     2341 ttctttaagg taaaaatcat tttggtagcc aagatagaat gaccacctaa atcgaaaaac
     2401 gaatcatctg gtgatacaga tgctggcttg gtaggtaata tacttaacca taagtctcta
     2461 acctcgcgct caacattggt aaactgagag tcgtcagttt cagaaactgt attttcagct
     2521 accaaattta attgcttggg agttgggaat tgaagtttag gcttatcaac tttaccattt
     2581 ggattcaatg gtagtttatc cataaccaca atcaaggaag gcatagcata gctagccaat
     2641 cttttcttta agaaagtcct gatgtccttg gataaaagat ggtaaccgat taagccctta
     2701 actatagggt cagtttcaac ctcctttgga acatcacttt ggaacttaga caagtcatct
     2761 ggcttgtcaa atcttgggac cataaatgtg atcaatgttg gctcattgtc ggcatttttg
     2821 cgaactaaag taatgttttc tcttaccaat ggatgttggg aaatgtgcgt atctatttct
     2881 cctaattcga ttctgaaccc acgaatttta acttgatcat cagccctacc gcaacattca
     2941 cagtcaccgt ttggtagata acgacctaaa tcacccgttc tgtacaatct atctcttgga
     3001 cctaaccaga attgtctcca aggttcacca ttatccttat ccaaataatt ccagtgatct
     3061 ttttcaacaa accagttgtt cacaaatttt tctttattca attctggtaa tcctctataa
     3121 ccttcggcca aaccacctgc acgaacataa atctcaccta tttcgccaat accacatatt
     3181 tgagtacggt cgttcctgtt aacaactaat agctgaacgt tcaacatacc tttaccagca
     3241 ggcatgacat ctttcaattt tttcaaaaag tttggatcgt cattttttga tttaacttcg
     3301 aaataagaaa ctgcacgctg tgtttcagta gtaccgtaca tattaacaat acggcaattt
     3361 tctgccaagg tttgtaacct cagacaatca cgttttgtta aaatgtcacc cacaaagaac
     3421 gcatgatgta acttagggaa tggtgtagta gcttgggcag taagtaattg acccatggca
     3481 ggtgttaaat gggtaactgt gcaaccatac ttactcatcc attccgctaa acggcccggt
     3541 gtaccaatat catcttgagt agggacatac aattgggcac ctaaaaataa tggtgtaaac
     3601 atatctcttt gaattggatc atgtgcaata ccgctcagca ttgtgaattt atcattttct
     3661 gttaagttga accttttgga catccaattg aaataataag ccaaggaaaa atgtctacca
     3721 agaacaccct taggaatacc ttcggaacca gatgtgaaag atagggttgg gttggaatct
     3781 ggtccaacta caacacctgt tctggtgtct ttgtagtgat cgtatggagc caaaacatcc
     3841 tcgccattgt ccaatttgcc accttcaata gtaccatttt cttgaatagc gatggaattg
     3901 attcttgaaa caatctccaa ttcatcattg atgtaatctt ctactaattg atccaattgt
     3961 ccagcagctc taataacaat caacccacgt ggtttagcaa cacctaagta aatggtttgt
     4021 ctggctgggg gatatgcagg gtcgataact gaaaaggttg cgccagcttt caagacaccc
     4081 atcacacata ccatcaaatc cacaccccta gaagaataga tcatcactac atcacctctt
     4141 ttgatacctg ttttaatcaa ataatgggca actatgttag aagtgcggtt gatgtcgcga
     4201 taagtgaaag aacgggactt gtcggaattt agtgttggag tctccacaac acaggttctc
     4261 tctgggaagg cttcagcatt gtcctggaaa atgtcgtgaa tacaccccac gaaatcgcac
     4321 cagcccaagt tcttagttgg atcaggtaaa ctatccttgg atgatgcggt gatcagagag
     4381 attttagtta tgcaattgga tggatcgctt agcgcagcag tcaaatattg agtaaattgg
     4441 tccgcaacaa tggttactct ttcattcgaa tacagtaagc tgttataaat taagttcaaa
     4501 acatgcgcat tattactggt atccaaattc aaagcaaagt ccactaaatg atgcttgaac
     4561 tcgtctaatt tgaaatcttg gttttccaaa aaggccaaac ggaacaactg aggggtcctt
     4621 tccaagtctt ggcaactttg aattttttca gctagctcgt caaaggaaaa attggcctca
     4681 atagaattga gcttgttcaa ctcattgtta attgtagaat acagctcatt aaatgaccac
     4741 gttggttgaa tattgaatct taagattttg ttattcgcaa tataaagaac aatatcatcg
     4801 tcaccagtta ctctatatat caatgcagcc catacactca aagcgacagc gtatttgtta
     4861 gaaaaactat catgaggcac atcgagctga ggtagctgta acgaatatgt agcttgtttc
     4921 gtataaggtt cttgttgtgg gcgtaaaaag tcatgtggta acactgaaag agttggatta
     4981 tccaacttct ctatccagac cttttcgtta gtcattagtg agtaactctg tgatatgtct
     5041 ctataattag cagtttttca ctgaaattcc aggaaaggta ataaactcag attttttttt
     5101 atactattgg ctgcttgtta cttatatatc ttgaacttct cccagcgggt cttcaaatac
     5161 atttgggcga tgttcatgtt cattaggcag gtatttcgac attgagtcac acgcgaaaaa
     5221 ccgccggaat tttttatgta attgcaagtg caattccgct ggcaaaacta ttgaagagtt
     5281 ttcctcgcta aaactgtgcg atgcctctcg atcggatgtg ctgcaaggcg attaagttgg
     5341 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac
     5401 gactcactat agggcgaatt ggagaaacag aataatatat aaacctgtat aatataacct
     5461 tgaagactat atttcttttc gcggccgcct atagggcgaa ttggagctcc accgcggtgg
     5521 ctctagaact agtggatccc ccgggctgca ggaattcgat atcaagctta tcgataccgt
     5581 cgacctcgag ggggggcccg gtacccagct tcggccgtat ttacagataa cgatgtcgtt
     5641 attatatata tatatatata ttgtcacttg tacccagctt ttgttccctt tagtgagggt
     5701 taatttcgag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5761 tcacaacagc tgagaagcgg tcagcaagaa gaaaatcgaa taatcagttt caatgaatat
     5821 agacgtaaca aaggcacaaa aaagaaaaca agtagatatg tcatacatat atatatattt
     5881 caggcagcaa gtgaccatca accaggaagt gtgaaatagc aaagaactgt ttacttcttc
     5941 agacttagaa agctcttccc ataggagaaa gcaactgctt tcctttgtag taattgattg
     6001 tcttttcagc ccttgacgca acaccgtccg ctgtgaaatc gaacaatttg taaatttcag
     6061 gcgcgccatc ctgcaggcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6121 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6181 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6241 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6301 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6361 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6421 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6481 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6541 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6601 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6661 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6721 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6781 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6841 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6901 aagatccttt gatcttttgt ttaaacctac ggggtctgac gctcagtgga acgaaaactc
     6961 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7021 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7081 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7141 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7201 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7261 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7321 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7381 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7441 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7501 tagctccttc ggtcctccga tggttgtcag aagtaagttg gccgcagtgt tatcactcat
     7561 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7621 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7681 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7741 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7801 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7861 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7921 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7981 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8041 gcgcacattt ccccgaaaag tgccacct
//
