LOCUS       sequence-108546-        5111 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 5111)
  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 5111)
  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: pRG215
FEATURES             Location/Qualifiers
     source          1..5111
                     /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(819..1913)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFQNHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1914..2319)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     2515..2537
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2529..2546
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2530..2546
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2553..2572
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2553..2571
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2680..2696
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2680..2696
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2730..2746)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2731..2747)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(2845..2863)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2884..2900)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2884..2900)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2897..2919)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2908..2924
                     /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(3183..3200)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3354..3942)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3434..3453)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4121..4912,4913..4981))
                     /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     4744..4763
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4982..5086)
                     /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 gcgcctaact ttttgtgtgg tgccctcctc cttgtcaata ttaatgttaa agtgcaattc
      601 tttttcctta tcacgttgag ccattagtat caatttgctt acctgtattc ctttacatcc
      661 tcctttttct ccttcttgat aaatgtatgt agattgcgta tatagtttcg tctaccctat
      721 gaacatattc cattttgtaa tttcgtgtcg catatgtttc tattatgaat ttcatttata
      781 aagtttatgt acaaatatca taaaaaaaga gaatcttttt aagcaaggat tttcttaact
      841 tcttcggcga cagcatcacc gacttcggtg gtactgttgg aaccacctaa atcaccagtt
      901 ctgatacctg catccaaaac ctttttaact gcatcttcaa tggccttacc ttcttcaggc
      961 aagttcaatg acaatttcaa catcattgca gcagacaaga tagtggcgat agggttgacc
     1021 ttattctttg gcaaatctgg agcagaaccg tggcatggtt cgtacaaacc aaatgcggtg
     1081 ttcttgtctg gcaaagaggc caaggacgca gatggcaaca aacccaagga acctgggata
     1141 acggaggctt catcggagat gatgtcacca aacatgttgc tggtgattat aataccattt
     1201 aggtgggttg ggttcttaac taggatcatg gcggcagaat caatcaattg atgttgaacc
     1261 ttcaatgtag gaaattcgtt cttgatggtt tcctccacag tttttctcca taatcttgaa
     1321 gaggccaaaa cattagcttt atccaaggac caaataggca atggtggctc atgttgtagg
     1381 gccatgaaag cggccattct tgtgattctt tgcacttctg gaacggtgta ttgttcacta
     1441 tcccaagcga caccatcacc atcgtcttcc tttctcttac caaagtaaat acctcccact
     1501 aattctctga caacaacgaa gtcagtacct ttagcaaatt gtggcttgat tggagataag
     1561 tctaaaagag agtcggatgc aaagttacat ggtcttaagt tggcgtacaa ttgaagttct
     1621 ttacggattt ttagtaaacc ttgttcaggt ctaacactac cagtacccca tttaggacca
     1681 cccacagcac ctaacaaaac ggcatcagcc ttcttggagg cttccagcgc ctcatctgga
     1741 agtggaacac ctgtagcatc gatagcagca ccaccaatta aatgattttg gaaatcgaac
     1801 ttgacattgg aacgaacatc agaaatagct ttaagaacct taatggcttc ggctgtgatt
     1861 tcttgaccaa cgtggtcacc tggcaaaacg acgatcttct taggggcaga cattagaatg
     1921 gtatatcctt gaaatatata tatatattgc tgaaatgtaa aaggtaagaa aagttagaaa
     1981 gtaagacgat tgctaaccac ctattggaaa aaacaatagg tccttaaata atattgtcaa
     2041 cttcaagtat tgtgatgcaa gcatttagtc atgaacgctt ctctattcta tatgaaaagc
     2101 cggttccggc gctctcacct ttcctttttc tcccaatttt tcagttgaaa aaggtatatg
     2161 cgtcaggcga cctctgaaat taacaaaaaa tttccagtca tcgaatttga ttctgtgcga
     2221 tagcgcccct gtgtgttctc gttatgttga ggaaaaaaat aatggttgct aagagattcg
     2281 aactcttgca tcttacgata cctgagtatt cccacagtta actgcggtca agatatttct
     2341 tgaatcaggc gccttagacc gctcggccaa acaaccaatt acttgttgag aaatagagta
     2401 taattatcct ataaatataa cgtttttgaa cacacatgaa caaggaagta caggacaatt
     2461 gattttcgat cggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     2521 cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg
     2581 agaaacagaa taatatataa acctgtataa tataaccttg aagactatat ttcttttcgc
     2641 ggccgcctat agggcgaatt ggagctccac cgcggtggct ctagaactag tggatccccc
     2701 gggctgcagg aattcgatat caagcttatc gataccgtcg acctcgaggg ggggcccggt
     2761 acccagcttc ggccgtattt acagataacg atgtcgttat tatatatata tatatatatt
     2821 gtcacttgta cccagctttt gttcccttta gtgagggtta atttcgagct tggcgtaatc
     2881 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaacagctg taatatatag
     2941 tagtaacctg aaaaatagtc aatgcccgtg gttcatttgc caatagaaca gtcgatgtga
     3001 tcaatgtaca cgaaagtcaa tataaaatat ccaaatagcc aatggtgtca ggacggctaa
     3061 ggcaaatgca actttgaaat acttattcgg tttgcatctt ggcgcgcctc ttcctgcagg
     3121 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3181 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3241 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3301 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3361 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3421 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3481 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3541 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3601 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3661 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3721 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3781 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3841 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3901 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3961 tgtttaaacc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4021 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4081 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4141 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4201 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4261 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4321 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4381 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4441 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4501 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4561 cgatggttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4621 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4681 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4741 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4801 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4861 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4921 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4981 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5041 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5101 aagtgccacc t
//
