LOCUS       sequence-108549-        4557 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 4557)
  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 4557)
  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: pRG216
FEATURES             Location/Qualifiers
     source          1..4557
                     /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(839..1642)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDLLTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSACVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1643..1859)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     1910..1932
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1924..1941
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1925..1941
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1948..1967
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1948..1966
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2075..2091
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2075..2091
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2125..2141)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2126..2142)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(2240..2258)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2279..2295)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2279..2295)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2292..2314)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2303..2319
                     /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(2629..2646)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2800..3388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2880..2899)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3567..4358,4359..4427))
                     /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     4190..4209
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4428..4532)
                     /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 taactctggc
      541 gcgccactac tttttggaac tcttgttgtt ctttggagtt caatgcgtcc atctttacag
      601 tcctgtctta ttgttcttga tttgtgcccc gtaaaatact gttacttggt tctggcgagg
      661 tattggatag ttccttttta taaaggccat gaaggttttt ctttccaatt tttttttttt
      721 cgtcattata gaaatcatta cgaccgagat tcccggcata tggtaataac tgatataatt
      781 aaattgaagc tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt
      841 agttttgctg gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac
      901 cctctacctt agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag
      961 atcctgtaga gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt
     1021 catctaaacc cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca
     1081 tgtctctttg agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag
     1141 tacccttagt atattctcca gtagctaggg agcccttgca tgacaattct gctaacatca
     1201 aaaggcctct aggttccttt gttacttctt ccgccgcctg cttcaaaccg ctaacaatac
     1261 ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac
     1321 acgcagagta ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga
     1381 gtaaaaaatt gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa
     1441 aatcagtcaa gagatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa
     1501 ctaactccag taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct
     1561 tttcgtgcat gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt
     1621 ccttatatgt agctttcgac atgatttatc ttcgtttcct gcacgttttt gttctgtgca
     1681 gttgggttaa gaatactggg caatttcatg tttcttcaac accacctatg cgtatatata
     1741 ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ctcggagatt accgaatcaa
     1801 aaaaatttca aagaaaccgg aatcaaaaaa aagaacaaaa aaaaaaaaga tgaatgaaaa
     1861 gcgatcggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     1921 cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga attggagaaa
     1981 cagaataata tataaacctg tataatataa ccttgaagac tatatttctt ttcgcggccg
     2041 cctatagggc gaattggagc tccaccgcgg tggctctaga actagtggat cccccgggct
     2101 gcaggaattc gatatcaagc ttatcgatac cgtcgacctc gagggggggc ccggtaccca
     2161 gcttcggccg tatttacaga taacgatgtc gttattatat atatatatat atattgtcac
     2221 ttgtacccag cttttgttcc ctttagtgag ggttaatttc gagcttggcg taatcatggt
     2281 catagctgtt tcctgtgtga aattgttatc cgctcacaac agctggatga attgaatagc
     2341 tttatggacc ctgaaaccac agccacatta accttctttg atggtcaaaa cttatccttc
     2401 accataaata tgcctcgcaa aaaaggtaat taacatatat agaattacat tatttatgaa
     2461 atatcatcac tatctcttag catctttaat ccttttctac atcagataac ttcggtttgt
     2521 tatcatcgtc tgtattgtca tcaattggcg cgcctctccc tgcaggcatt aatgaatcgg
     2581 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2641 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2701 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2761 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2821 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2881 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2941 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3001 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3061 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3121 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3181 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3241 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3301 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3361 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttgtt taaacctacg
     3421 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3481 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3541 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3601 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3661 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3721 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3781 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3841 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3901 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3961 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat ggttgtcaga
     4021 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4081 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4141 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4201 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4261 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4321 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4381 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4441 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4501 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacct
//
