LOCUS       sequence-108550-        5384 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Episomal E. coli/S. cerevisiae shuttle vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5384)
  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 5384)
  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: pRG226
FEATURES             Location/Qualifiers
     source          1..5384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..1349
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(970..1017)
                     /label=FRT (mutant)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FRT site with a mutation in the core sequence (Azam 
                     et al., 1997)"
     protein_bind    complement(970..1017)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(1666..2469)
                     /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(2470..2686)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     2737..2759
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2751..2768
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2752..2768
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2775..2794
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2775..2793
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2902..2918
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2902..2918
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2952..2968)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2953..2969)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(3067..3085)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3106..3122)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3106..3122)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3119..3141)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3130..3146
                     /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(3456..3473)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3627..4215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3707..3726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4394..5185,5186..5254))
                     /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     5017..5036
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5255..5359)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta
       61 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg
      121 ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag
      181 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
      241 gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc
      301 gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct
      361 ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta
      421 ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga
      481 ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat tatattctat
      541 accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt
      601 ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg
      661 tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg
      721 tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt
      781 tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca
      841 aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt
      901 acagtccggt gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt tggttttcaa
      961 aagcgctctg aagttcctat actttttaga gaataggaac ttcggaatag gaacttcaaa
     1021 gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat acagctcact
     1081 gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat atatacatga gaagaacggc
     1141 atagtgcgtg tttatgctta aatgcgtact tatatgcgtc tatttatgta ggatgaaagg
     1201 tagtctagta cctcctgtga tattatccca ttccatgcgg ggtatcgtat gcttccttca
     1261 gcactaccct ttagctgttc tatatgctgc cactcctcaa ttggattagt ctcatccttc
     1321 aatgctatca tttcctttga tattggatca tattaattaa ctctggcgcg ccactacttt
     1381 ttggaactct tgttgttctt tggagttcaa tgcgtccatc tttacagtcc tgtcttattg
     1441 ttcttgattt gtgccccgta aaatactgtt acttggttct ggcgaggtat tggatagttc
     1501 ctttttataa aggccatgaa ggtttttctt tccaattttt tttttttcgt cattatagaa
     1561 atcattacga ccgagattcc cggcatatgg taataactga tataattaaa ttgaagctct
     1621 aatttgtgag tttagtatac atgcatttac ttataataca gttttttagt tttgctggcc
     1681 gcatcttctc aaatatgctt cccagcctgc ttttctgtaa cgttcaccct ctaccttagc
     1741 atcccttccc tttgcaaata gtcctcttcc aacaataata atgtcagatc ctgtagagac
     1801 cacatcatcc acggttctat actgttgacc caatgcgtct cccttgtcat ctaaacccac
     1861 accgggtgtc ataatcaacc aatcgtaacc ttcatctctt ccacccatgt ctctttgagc
     1921 aataaagccg ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac ccttagtata
     1981 ttctccagta gctagggagc ccttgcatga caattctgct aacatcaaaa ggcctctagg
     2041 ttcctttgtt acttcttccg ccgcctgctt caaaccgcta acaatacctg ggcccaccac
     2101 accgtgtgca ttcgtaatgt ctgcccattc tgctattctg tatacacacg cagagtactg
     2161 caatttgact gtattaccaa tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta
     2221 cttggcggat aatgccttta gcggcttaac tgtgccctcc atggaaaaat cagtcaagag
     2281 atccacatgt gtttttagta aacaaatttt gggacctaat gcttcaacta actccagtaa
     2341 ttccttggtg gtacgaacat ccaatgaagc acacaagttt gtttgctttt cgtgcatgat
     2401 attaaatagc ttggcagcaa caggactagg atgagtagca gcacgttcct tatatgtagc
     2461 tttcgacatg atttatcttc gtttcctgca cgtttttgtt ctgtgcagtt gggttaagaa
     2521 tactgggcaa tttcatgttt cttcaacacc acctatgcgt atatatacca atctaagtct
     2581 gtgctccttc cttcgttctt ccttctgctc ggagattacc gaatcaaaaa aatttcaaag
     2641 aaaccggaat caaaaaaaag aacaaaaaaa aaaaagatga atgaaaagcg atcggatgtg
     2701 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     2761 cggccagtga attgtaatac gactcactat agggcgaatt ggagaaacag aataatatat
     2821 aaacctgtat aatataacct tgaagactat atttcttttc gcggccgcct atagggcgaa
     2881 ttggagctcc accgcggtgg ctctagaact agtggatccc ccgggctgca ggaattcgat
     2941 atcaagctta tcgataccgt cgacctcgag ggggggcccg gtacccagct tcggccgtat
     3001 ttacagataa cgatgtcgtt attatatata tatatatata ttgtcacttg tacccagctt
     3061 ttgttccctt tagtgagggt taatttcgag cttggcgtaa tcatggtcat agctgtttcc
     3121 tgtgtgaaat tgttatccgc tcacaacagc tggatgaatt gaatagcttt atggaccctg
     3181 aaaccacagc cacattaacc ttctttgatg gtcaaaactt atccttcacc ataaatatgc
     3241 ctcgcaaaaa aggtaattaa catatataga attacattat ttatgaaata tcatcactat
     3301 ctcttagcat ctttaatcct tttctacatc agataacttc ggtttgttat catcgtctgt
     3361 attgtcatca attggcgcgc ctctccctgc aggcattaat gaatcggcca acgcgcgggg
     3421 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3481 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3541 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3601 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3661 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3721 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3781 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3841 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3901 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3961 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4021 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4081 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4141 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4201 aaaaaaggat ctcaagaaga tcctttgatc ttttgtttaa acctacgggg tctgacgctc
     4261 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4321 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4381 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4441 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4501 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4561 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4621 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4681 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4741 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4801 tgtgcaaaaa agcggttagc tccttcggtc ctccgatggt tgtcagaagt aagttggccg
     4861 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4921 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4981 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5041 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5101 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5161 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5221 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5281 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5341 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acct
//
