LOCUS       sequence-108554-        4023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative E. coli/S. cerevisiae shuttle vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4023)
  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 4023)
  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: pRG235
FEATURES             Location/Qualifiers
     source          1..4023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(287..1381)
                     /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"
     primer_bind     1459..1481
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1473..1490
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1474..1490
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1497..1516
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1497..1515
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1624..1640
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1624..1640
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1674..1690)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1675..1691)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(1789..1807)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1828..1844)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1828..1844)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1841..1863)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1852..1868
                     /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(2095..2112)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2266..2854)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2346..2365)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3033..3824,3825..3893))
                     /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     3656..3675
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3894..3998)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgaaa gaacgcggca ttagcaacat ttgcgaagcg aacgattttg cgaactgcta
       61 ttaattaact ctggcgcgcc actatcgtct atcaactaat agttatatta tcaatatatt
      121 atcatatacg gtgttaagat gatgacataa gttatgagaa gctgtcatcg aagttagagg
      181 aagctgaagt gcaaggattg ataatgtaat aggataatca tatgtttcta ttatgaattt
      241 catttataaa gtttatgtac aaatatcata aaaaaagaga atctttttaa gcaaggattt
      301 tcttaacttc ttcggcgaca gcatcaccga cttcggtggt actgttggaa ccacctaaat
      361 caccagttct gatacctgca tccaaaacct ttttaactgc atcttcaatg gccttacctt
      421 cttcaggcaa gttcaatgac aatttcaaca tcattgcagc agacaagata gtggcgatag
      481 ggttgacctt attctttggc aaatctggag cagaaccgtg gcatggttcg tacaaaccaa
      541 atgcggtgtt cttgtctggc aaagaggcca aggacgcaga tggcaacaaa cccaaggaac
      601 ctgggataac ggaggcttca tcggagatga tgtcaccaaa catgttgctg gtgattataa
      661 taccatttag gtgggttggg ttcttaacta ggatcatggc ggcagaatca atcaattgat
      721 gttgaacctt caatgtagga aattcgttct tgatggtttc ctccacagtt tttctccata
      781 atcttgaaga ggccaaaaca ttagctttat ccaaggacca aataggcaat ggtggctcat
      841 gttgtagggc catgaaagcg gccattcttg tgattctttg cacttctgga acggtgtatt
      901 gttcactatc ccaagcgaca ccatcaccat cgtcttcctt tctcttacca aagtaaatac
      961 ctcccactaa ttctctgaca acaacgaagt cagtaccttt agcaaattgt ggcttgattg
     1021 gagataagtc taaaagagag tcggatgcaa agttacatgg tcttaagttg gcgtacaatt
     1081 gaagttcttt acggattttt agtaaacctt gttcaggtct aacactacca gtaccccatt
     1141 taggaccacc cacagcacct aacaaaacgg catcagcctt cttggaggct tccagcgcct
     1201 catctggaag tggaacacct gtagcatcga tagcagcacc accaattaaa tgattttgga
     1261 aatcgaactt gacattggaa cgaacatcag aaatagcttt aagaacctta atggcttcgg
     1321 ctgtgatttc ttgaccaacg tggtcacctg gcaaaacgac gatcttctta ggggcagaca
     1381 ttagaatggt atatccttga aatatatata cgatcggatg tgctgcaagg cgattaagtt
     1441 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgtaat
     1501 acgactcact atagggcgaa ttggagaaac agaataatat ataaacctgt ataatataac
     1561 cttgaagact atatttcttt tcgcggccgc ctatagggcg aattggagct ccaccgcggt
     1621 ggctctagaa ctagtggatc ccccgggctg caggaattcg atatcaagct tatcgatacc
     1681 gtcgacctcg agggggggcc cggtacccag cttcggccgt atttacagat aacgatgtcg
     1741 ttattatata tatatatata tattgtcact tgtacccagc ttttgttccc tttagtgagg
     1801 gttaatttcg agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     1861 gctcacaaca gctggaaaca tataaaacgg aatgaggaat aatcgtaata ttagtatgta
     1921 gaaatataga ttccattttg aggattccta tatccttgag gagaacttct agtatattct
     1981 gtatacctaa tattatagcc tttatcaaca atggcgcgcc tctccctgca ggcattaatg
     2041 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2101 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2161 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2221 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2281 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2341 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2401 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2461 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2521 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2581 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2641 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2701 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2761 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2821 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttgtttaaa
     2881 cctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2941 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3001 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3061 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3121 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3181 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3241 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3301 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3361 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3421 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatggtt
     3481 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3541 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3601 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3661 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3721 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3781 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3841 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3901 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3961 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4021 cct
//
