LOCUS       sequence-108545-        4646 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 4646)
  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 4646)
  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: pRG205
FEATURES             Location/Qualifiers
     source          1..4646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(354..1448)
                     /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(1449..1854)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     2050..2072
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2064..2081
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2065..2081
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2088..2107
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2088..2106
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2215..2231
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2215..2231
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2265..2281)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2266..2282)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(2380..2398)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2419..2435)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2419..2435)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2432..2454)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2443..2459
                     /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(2718..2735)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2889..3477)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2969..2988)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3656..4447,4448..4516))
                     /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     4279..4298
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4517..4621)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgaaa gaacgcggca ttagcaacat ttgcgaagcg aacgattttg cgaactgcta
       61 ttaattaact ctggcgcgcc taactttttg tgtggtgccc tcctccttgt caatattaat
      121 gttaaagtgc aattcttttt ccttatcacg ttgagccatt agtatcaatt tgcttacctg
      181 tattccttta catcctcctt tttctccttc ttgataaatg tatgtagatt gcgtatatag
      241 tttcgtctac cctatgaaca tattccattt tgtaatttcg tgtcgcatat gtttctatta
      301 tgaatttcat ttataaagtt tatgtacaaa tatcataaaa aaagagaatc tttttaagca
      361 aggattttct taacttcttc ggcgacagca tcaccgactt cggtggtact gttggaacca
      421 cctaaatcac cagttctgat acctgcatcc aaaacctttt taactgcatc ttcaatggcc
      481 ttaccttctt caggcaagtt caatgacaat ttcaacatca ttgcagcaga caagatagtg
      541 gcgatagggt tgaccttatt ctttggcaaa tctggagcag aaccgtggca tggttcgtac
      601 aaaccaaatg cggtgttctt gtctggcaaa gaggccaagg acgcagatgg caacaaaccc
      661 aaggaacctg ggataacgga ggcttcatcg gagatgatgt caccaaacat gttgctggtg
      721 attataatac catttaggtg ggttgggttc ttaactagga tcatggcggc agaatcaatc
      781 aattgatgtt gaaccttcaa tgtaggaaat tcgttcttga tggtttcctc cacagttttt
      841 ctccataatc ttgaagaggc caaaacatta gctttatcca aggaccaaat aggcaatggt
      901 ggctcatgtt gtagggccat gaaagcggcc attcttgtga ttctttgcac ttctggaacg
      961 gtgtattgtt cactatccca agcgacacca tcaccatcgt cttcctttct cttaccaaag
     1021 taaatacctc ccactaattc tctgacaaca acgaagtcag tacctttagc aaattgtggc
     1081 ttgattggag ataagtctaa aagagagtcg gatgcaaagt tacatggtct taagttggcg
     1141 tacaattgaa gttctttacg gatttttagt aaaccttgtt caggtctaac actaccagta
     1201 ccccatttag gaccacccac agcacctaac aaaacggcat cagccttctt ggaggcttcc
     1261 agcgcctcat ctggaagtgg aacacctgta gcatcgatag cagcaccacc aattaaatga
     1321 ttttggaaat cgaacttgac attggaacga acatcagaaa tagctttaag aaccttaatg
     1381 gcttcggctg tgatttcttg accaacgtgg tcacctggca aaacgacgat cttcttaggg
     1441 gcagacatta gaatggtata tccttgaaat atatatatat attgctgaaa tgtaaaaggt
     1501 aagaaaagtt agaaagtaag acgattgcta accacctatt ggaaaaaaca ataggtcctt
     1561 aaataatatt gtcaacttca agtattgtga tgcaagcatt tagtcatgaa cgcttctcta
     1621 ttctatatga aaagccggtt ccggcgctct cacctttcct ttttctccca atttttcagt
     1681 tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca aaaaatttcc agtcatcgaa
     1741 tttgattctg tgcgatagcg cccctgtgtg ttctcgttat gttgaggaaa aaaataatgg
     1801 ttgctaagag attcgaactc ttgcatctta cgatacctga gtattcccac agttaactgc
     1861 ggtcaagata tttcttgaat caggcgcctt agaccgctcg gccaaacaac caattacttg
     1921 ttgagaaata gagtataatt atcctataaa tataacgttt ttgaacacac atgaacaagg
     1981 aagtacagga caattgattt tcgatcggat gtgctgcaag gcgattaagt tgggtaacgc
     2041 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac
     2101 tatagggcga attggagaaa cagaataata tataaacctg tataatataa ccttgaagac
     2161 tatatttctt ttcgcggccg cctatagggc gaattggagc tccaccgcgg tggctctaga
     2221 actagtggat cccccgggct gcaggaattc gatatcaagc ttatcgatac cgtcgacctc
     2281 gagggggggc ccggtaccca gcttcggccg tatttacaga taacgatgtc gttattatat
     2341 atatatatat atattgtcac ttgtacccag cttttgttcc ctttagtgag ggttaatttc
     2401 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaac
     2461 agctgtaata tatagtagta acctgaaaaa tagtcaatgc ccgtggttca tttgccaata
     2521 gaacagtcga tgtgatcaat gtacacgaaa gtcaatataa aatatccaaa tagccaatgg
     2581 tgtcaggacg gctaaggcaa atgcaacttt gaaatactta ttcggtttgc atcttggcgc
     2641 gcctcttcct gcaggcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2701 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2761 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2821 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2881 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2941 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3001 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3061 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3121 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3181 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3241 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3301 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3361 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3421 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3481 gatcctttga tcttttgttt aaacctacgg ggtctgacgc tcagtggaac gaaaactcac
     3541 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3601 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3661 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3721 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3781 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3841 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3901 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3961 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4021 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4081 gctccttcgg tcctccgatg gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4141 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4201 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4261 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4321 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4381 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4441 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4501 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4561 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4621 gcacatttcc ccgaaaagtg ccacct
//
