LOCUS       sequence-108543-        4298 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 4298)
  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 4298)
  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: pRG213MX
FEATURES             Location/Qualifiers
     source          1..4298
                     /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             738..1388
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQK
                     GEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQA
                     MGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLYS
                     FSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     primer_bind     1727..1749
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1741..1758
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1742..1758
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1765..1784
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1765..1783
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1892..1908
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1892..1908
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1942..1958)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1943..1959)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(2057..2075)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2096..2112)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2096..2112)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2109..2131)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2120..2136
                     /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(2370..2387)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2541..3129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2621..2640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3308..4099,4100..4168))
                     /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     3931..3950
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4169..4273)
                     /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 gcgcctcctc tagtacactc tatatttttt tatgcctcgg taatgatttt catttttttt
      601 tttccaccta gcggatgact cttttttttt cttagcgatt ggcattatca cataatgaat
      661 tatacattat ataaagtaat gtgatttctt cgaagaatat actaaaaaat gagcaggcaa
      721 gataaacgaa ggcacatatg aggagggctt ttgtagaaag aaatacgaac gaaacgaaaa
      781 tcagcgttgc catcgctttg gacaaagctc ccttacctga agagtcgaat tttattgatg
      841 aacttataac ttccaagcat gcaaaccaaa agggagaaca agtaatccaa gtagacacgg
      901 gaattggatt cttggatcac atgtatcatg cactggctaa acatgcaggc tggagcttac
      961 gactttactc aagaggtgat ttaatcatcg atgatcatca cactgcagaa gatactgcta
     1021 ttgcacttgg tattgcattc aagcaggcta tgggtaactt tgccggtgtt aaaagatttg
     1081 gacatgctta ttgtccactt gacgaagctc tttccagaag cgtagttgac ttgtcgggac
     1141 ggccctatgc tgttatcgat ttgggattaa agcgtgaaaa ggttggggaa ttgtcctgtg
     1201 aaatgatccc tcacttacta tattcctttt cggtagctgc tggaattact ttgcatgtta
     1261 cctgcttata tggtagtaat gaccatcatc gtgctgaaag cgcttttaaa tctctggctg
     1321 ttgccatgcg cgcggctact tctcttactg gaagttctga agtcccaagc acgaagggag
     1381 tgttgtaaag atgaattgga ttatgtcagg aaaagaacga caattttgca tccaaattgt
     1441 ctaaatttta gagttgcttg aaaacaatag aaccttactt gctttataat tacgttaatt
     1501 agaagcgtta tctcgtgaag gaatatagta cgtagccgta taaattgaat tgaatgttca
     1561 gcttatagaa tagagacact ttgctgttca atgcgtcgtc acttaccata ctcactttat
     1621 tatacgactt taagtataaa ctccgcggtt atggtaaaat taatgatgca caaacgtccg
     1681 atcggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     1741 tgtaaaacga cggccagtga attgtaatac gactcactat agggcgaatt ggagaaacag
     1801 aataatatat aaacctgtat aatataacct tgaagactat atttcttttc gcggccgcct
     1861 atagggcgaa ttggagctcc accgcggtgg ctctagaact agtggatccc ccgggctgca
     1921 ggaattcgat atcaagctta tcgataccgt cgacctcgag ggggggcccg gtacccagct
     1981 tcggccgtat ttacagataa cgatgtcgtt attatatata tatatatata ttgtcacttg
     2041 tacccagctt ttgttccctt tagtgagggt taatttcgag cttggcgtaa tcatggtcat
     2101 agctgtttcc tgtgtgaaat tgttatccgc tcacaacagc tgacaccgat tatttaaagc
     2161 tgcagcatac gatatatata catgtgtata tatgtatacc tatgaatgtc agtaagtatg
     2221 tatacgaaca gtatgatact gaagatgaca aggtaatgca tcattctata cgtgtcattc
     2281 tgaacgaggc gcgcctctcc ctgcaggcat taatgaatcg gccaacgcgc ggggagaggc
     2341 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2401 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2461 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2521 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2581 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2641 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2701 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2761 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2821 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2881 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2941 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3001 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3061 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3121 ggatctcaag aagatccttt gatcttttgt ttaaacctac ggggtctgac gctcagtgga
     3181 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3241 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3301 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3361 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3421 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3481 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3541 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3601 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3661 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3721 aaaaagcggt tagctccttc ggtcctccga tggttgtcag aagtaagttg gccgcagtgt
     3781 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3841 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3901 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3961 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4021 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4081 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4141 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4201 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4261 taggggttcc gcgcacattt ccccgaaaag tgccacct
//
