LOCUS       sequence-108548-        4092 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 4092)
  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 4092)
  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: pRG206
FEATURES             Location/Qualifiers
     source          1..4092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(374..1177)
                     /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(1178..1394)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     1445..1467
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1459..1476
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1460..1476
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1483..1502
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1483..1501
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1610..1626
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1610..1626
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1660..1676)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1661..1677)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(1775..1793)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1814..1830)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1814..1830)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1827..1849)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1838..1854
                     /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(2164..2181)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2335..2923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2415..2434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3102..3893,3894..3962))
                     /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     3725..3744
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3963..4067)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgaaa gaacgcggca ttagcaacat ttgcgaagcg aacgattttg cgaactgcta
       61 ttaattaact ctggcgcgcc actacttttt ggaactcttg ttgttctttg gagttcaatg
      121 cgtccatctt tacagtcctg tcttattgtt cttgatttgt gccccgtaaa atactgttac
      181 ttggttctgg cgaggtattg gatagttcct ttttataaag gccatgaagg tttttctttc
      241 caattttttt tttttcgtca ttatagaaat cattacgacc gagattcccg gcatatggta
      301 ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt
      361 ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt
      421 ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa
      481 caataataat gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca
      541 atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt
      601 catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct
      661 tcgcaatgtc aacagtaccc ttagtatatt ctccagtagc tagggagccc ttgcatgaca
      721 attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttccgcc gcctgcttca
      781 aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg
      841 ctattctgta tacacacgca gagtactgca atttgactgt attaccaatg tcagcaaatt
      901 ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg
      961 tgccctccat ggaaaaatca gtcaagagat ccacatgtgt ttttagtaaa caaattttgg
     1021 gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac
     1081 acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat
     1141 gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcacg
     1201 tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacaccac
     1261 ctatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgctcgg
     1321 agattaccga atcaaaaaaa tttcaaagaa accggaatca aaaaaaagaa caaaaaaaaa
     1381 aaagatgaat gaaaagcgat cggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1441 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag
     1501 ggcgaattgg agaaacagaa taatatataa acctgtataa tataaccttg aagactatat
     1561 ttcttttcgc ggccgcctat agggcgaatt ggagctccac cgcggtggct ctagaactag
     1621 tggatccccc gggctgcagg aattcgatat caagcttatc gataccgtcg acctcgaggg
     1681 ggggcccggt acccagcttc ggccgtattt acagataacg atgtcgttat tatatatata
     1741 tatatatatt gtcacttgta cccagctttt gttcccttta gtgagggtta atttcgagct
     1801 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaacagctg
     1861 gatgaattga atagctttat ggaccctgaa accacagcca cattaacctt ctttgatggt
     1921 caaaacttat ccttcaccat aaatatgcct cgcaaaaaag gtaattaaca tatatagaat
     1981 tacattattt atgaaatatc atcactatct cttagcatct ttaatccttt tctacatcag
     2041 ataacttcgg tttgttatca tcgtctgtat tgtcatcaat tggcgcgcct ctccctgcag
     2101 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2161 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2221 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2281 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2341 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2401 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2461 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2521 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2581 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2641 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2701 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2761 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2821 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2881 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2941 ttgtttaaac ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3001 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3061 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3121 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3181 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3241 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3301 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3361 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3421 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3481 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3541 ccgatggttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3601 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3661 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3721 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3781 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3841 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3901 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3961 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4021 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4081 aaagtgccac ct
//
