LOCUS       sequence-108555-        3763 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 3763)
  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 3763)
  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: pRG236
FEATURES             Location/Qualifiers
     source          1..3763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(301..1104)
                     /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"
     primer_bind     1199..1221
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1213..1230
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1214..1230
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1237..1256
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1237..1255
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1364..1380
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1364..1380
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1414..1430)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1415..1431)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(1529..1547)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1568..1584)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1568..1584)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1581..1603)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1592..1608
                     /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(1835..1852)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2006..2594)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2086..2105)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2773..3564,3565..3633))
                     /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     3396..3415
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3634..3738)
                     /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 tatggtaata actgatataa
      241 ttaaattgaa gctctaattt gtgagtttag tatacatgca tttacttata atacagtttt
      301 ttagttttgc tggccgcatc ttctcaaata tgcttcccag cctgcttttc tgtaacgttc
      361 accctctacc ttagcatccc ttccctttgc aaatagtcct cttccaacaa taataatgtc
      421 agatcctgta gagaccacat catccacggt tctatactgt tgacccaatg cgtctccctt
      481 gtcatctaaa cccacaccgg gtgtcataat caaccaatcg taaccttcat ctcttccacc
      541 catgtctctt tgagcaataa agccgataac aaaatctttg tcgctcttcg caatgtcaac
      601 agtaccctta gtatattctc cagtagctag ggagcccttg catgacaatt ctgctaacat
      661 caaaaggcct ctaggttcct ttgttacttc ttccgccgcc tgcttcaaac cgctaacaat
      721 acctgggccc accacaccgt gtgcattcgt aatgtctgcc cattctgcta ttctgtatac
      781 acacgcagag tactgcaatt tgactgtatt accaatgtca gcaaattttc tgtcttcgaa
      841 gagtaaaaaa ttgtacttgg cggataatgc ctttagcggc ttaactgtgc cctccatgga
      901 aaaatcagtc aagagatcca catgtgtttt tagtaaacaa attttgggac ctaatgcttc
      961 aactaactcc agtaattcct tggtggtacg aacatccaat gaagcacaca agtttgtttg
     1021 cttttcgtgc atgatattaa atagcttggc agcaacagga ctaggatgag tagcagcacg
     1081 ttccttatat gtagctttcg acatgattta tcttcgtttc ctgcacgttt ttgttctgtg
     1141 cagttgggtt cgatcggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     1201 cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa
     1261 ttggagaaac agaataatat ataaacctgt ataatataac cttgaagact atatttcttt
     1321 tcgcggccgc ctatagggcg aattggagct ccaccgcggt ggctctagaa ctagtggatc
     1381 ccccgggctg caggaattcg atatcaagct tatcgatacc gtcgacctcg agggggggcc
     1441 cggtacccag cttcggccgt atttacagat aacgatgtcg ttattatata tatatatata
     1501 tattgtcact tgtacccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt
     1561 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaaca gctggaaaca
     1621 tataaaacgg aatgaggaat aatcgtaata ttagtatgta gaaatataga ttccattttg
     1681 aggattccta tatccttgag gagaacttct agtatattct gtatacctaa tattatagcc
     1741 tttatcaaca atggcgcgcc tctccctgca ggcattaatg aatcggccaa cgcgcgggga
     1801 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     1861 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1921 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1981 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2041 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2101 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2161 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2221 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2281 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2341 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2401 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2461 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2521 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2581 aaaaaggatc tcaagaagat cctttgatct tttgtttaaa cctacggggt ctgacgctca
     2641 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2701 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2761 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2821 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2881 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     2941 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3001 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3061 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3121 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3181 gtgcaaaaaa gcggttagct ccttcggtcc tccgatggtt gtcagaagta agttggccgc
     3241 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3301 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3361 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3421 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3481 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3541 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3601 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3661 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3721 acaaataggg gttccgcgca catttccccg aaaagtgcca cct
//
