LOCUS       sequence-108540-        5829 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 5829)
  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 5829)
  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: pRG211
FEATURES             Location/Qualifiers
     source          1..5829
                     /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"
     promoter        581..931
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     CDS             932..2266
                     /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"
     primer_bind     2939..2961
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2953..2970
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2954..2970
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2977..2996
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2977..2995
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3104..3120
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3104..3120
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3154..3170)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3155..3171)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(3269..3287)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3308..3324)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3308..3324)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3321..3343)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3332..3348
                     /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(3901..3918)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4072..4660)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4152..4171)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4839..5630,5631..5699))
                     /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     5462..5481
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5700..5804)
                     /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 taaacacaca
      541 ccataccttc ccatatgtcc tggcgcgccc ttagctctca ttattttttg ctttttctct
      601 tgaggtcaca tgatcgcaaa atggcaaatg gcacgtgaag ctgtcgatat tggggaactg
      661 tggtggttgg caaatgacta attaagttag tcaaggcgcc atcctcatga aaactgtgta
      721 acataataac cgaagtgtcg aaaaggtggc accttgtcca attgaacacg ctcgatgaaa
      781 aaaataagat atatataagg ttaagtaaag cgtctgttag aaaggaagtt tttccttttt
      841 cttgctctct tgtcttttca tctactattt ccttcgtgta atacagggtc gtcagataca
      901 tagatacaat tctattaccc ccatccatac aatgccatct catttcgata ctgttcaact
      961 acacgccggt caagagaacc ctggtgacaa tgctcacaga tccagagctg taccaattta
     1021 cgccaccact tcttatgttt tcgaaaactc taagcatggt tcgcaattgt ttggtttaga
     1081 agttccaggt tacgtctatt cccgtttcca aaacccaacc agtaatgttt tggaagaaag
     1141 aattgctgct ttagaaggtg gtgctgctgc tttggctgtt tcctccggtc aagccgctca
     1201 aacccttgcc atccaaggtt tggcacacac tggtgacaac atcgtttcca cttcttactt
     1261 atacggtggt acttataacc agttcaaaat ctcgttcaaa agatttggta tcgaggctag
     1321 atttgttgaa ggtgacaatc cagaagaatt tgaaaaggtc tttgatgaaa gaaccaaggc
     1381 tgtttatttg gaaaccattg gtaatccaaa gtacaatgtt ccggattttg aaaaaattgt
     1441 tgcaattgct cacaaacacg gtattccagt tgtcgttgac aacacatttg gtgccggtgg
     1501 ttacttctgt cagccaatta aatacggtgc tgatattgta acacattctg ctaccaaatg
     1561 gattggtggt catggtacta ctatcggtgg tattattgtt gactctggta agttcccatg
     1621 gaaggactac ccagaaaagt tccctcaatt ctctcaacct gccgaaggtt atcacggtac
     1681 tatctacaat gaagcctacg gtaacttggc atacatcgtt catgttagaa ctgaactatt
     1741 aagagatttg ggtccattga tgaacccatt tgcctctttc ttgctactac aaggtgttga
     1801 aacattatct ttgagagctg aaagacacgg tgaaaatgca ttgaagttag ccaaatggtt
     1861 agaacaatcc ccatacgtat cttgggtttc ataccctggt ttagcatctc attctcatca
     1921 tgaaaatgct aagaagtatc tatctaacgg tttcggtggt gtcttatctt tcggtgtaaa
     1981 agacttacca aatgccgaca aggaaactga cccattcaaa ctttctggtg ctcaagttgt
     2041 tgacaattta aagctagcct ctaacttggc caatgttggt gatgccaaga ccttagtcat
     2101 tgctccatac ttcactaccc acaaacaatt aaatgacaaa gaaaagttgg catctggtgt
     2161 taccaaggac ttaattcgtg tctctgttgg tatcgaattt attgatgaca ttattgcaga
     2221 cttccagcaa tcttttgaaa ctgttttcgc tggccaaaaa ccatgagtgt gcgtaatgag
     2281 ttgtaaaatt atgtataaac ctactttctc tcacaagtac tatactttta taaaacgaac
     2341 tttattgaaa tgaatatcct ttttttccct tgttacatgt cgtgactcgt actttgaacc
     2401 taaattgttc taacatcaaa gaacagtgtt aattcgcagt cgagaagaaa aatatggtga
     2461 acaagactca tctacttcat gagactactt tacgcctcct ataaagctgt cacactggat
     2521 aaatttattg taggaccaag ttacaaaaga ggatgatgga ggtttcttta caataaagaa
     2581 gcacatgtgt gttaacgttt ttagtatttg cttgttatgt aaatcaggaa aacttcgcgg
     2641 gatttggttg gatgctactt tccatacaat aaatattata gatcaaaaaa gccaaattac
     2701 aagtaaagat tagtaaagct gttgcaattc catcgttgat aaaaatgtta gttattaaat
     2761 ataaaagtca gaataggtga acttggattt aattgttggc atttcgttgc tgctagaggc
     2821 cataatatta gatagccagg acatactagg tctcctcgtg gtataggaat ccataaaatg
     2881 gaattggtga cgatcggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     2941 cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa
     3001 ttggagaaac agaataatat ataaacctgt ataatataac cttgaagact atatttcttt
     3061 tcgcggccgc ctatagggcg aattggagct ccaccgcggt ggctctagaa ctagtggatc
     3121 ccccgggctg caggaattcg atatcaagct tatcgatacc gtcgacctcg agggggggcc
     3181 cggtacccag cttcggccgt atttacagat aacgatgtcg ttattatata tatatatata
     3241 tattgtcact tgtacccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt
     3301 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaaca gctgtcagat
     3361 atagtcggat tgccctttta agcaatcaat agtgttttat ttgcaacaat gtcgtcatag
     3421 tttaatatgt cctataagat gttaacttgc tcaacattca acaaagtttg gtctcttggc
     3481 ccagttggtt aaggcaccgt gctaataacg cggggatcag cggttcgatc ccgctagaga
     3541 ccatttattt ttaaattcga ccgtctgaca gacggtgcac ttattactta attttttttt
     3601 ttttttaatc ctcgttaaac tgagtacata atcagcaaca atataatata aaagttttgc
     3661 ttttttatgt acgtacaata ggggtactta aataaataaa taaacaataa aacaatcaat
     3721 aaataaataa tatacgatca taaattacga gatatatttc atagtcttga cttctccagc
     3781 cgaacgtcat taaatttaag gcagaaaaga taaacaaggg cgcgccagag cctgcaggca
     3841 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3901 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3961 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4021 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4081 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4141 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4201 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4261 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4321 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4381 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4441 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4501 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4561 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4621 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttg
     4681 tttaaaccta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4741 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4801 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4861 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4921 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4981 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5041 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5101 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5161 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5221 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5281 atggttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5341 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5401 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5461 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5521 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5581 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5641 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5701 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5761 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5821 gtgccacct
//
