LOCUS       sequence-108385-        5364 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 5364)
  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 5364)
  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: pRG201
FEATURES             Location/Qualifiers
     source          1..5364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        116..466
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     CDS             467..1801
                     /codon_start=1
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /product="O-acetylserine and O-acetylhomoserine 
                     sulfhydrylase, required for sulfur amino acid synthesis"
                     /label=MET17
                     /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ
                     LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI
                     VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV
                     PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII
                     VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA
                     SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG
                     FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
     primer_bind     2474..2496
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2488..2505
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2489..2505
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2512..2531
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2512..2530
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2639..2655
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2639..2655
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2689..2705)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2690..2706)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(2804..2822)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2843..2859)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2843..2859)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2856..2878)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2867..2883
                     /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(3436..3453)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3607..4195)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3687..3706)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4374..5165,5166..5234))
                     /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     4997..5016
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5235..5339)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgaaa gaacgcggca ttagcaacat ttgcgaagcg aacgattttg cgaactgcta
       61 ttaattaaac acacaccata ccttcccata tgtcctggcg cgcccttagc tctcattatt
      121 ttttgctttt tctcttgagg tcacatgatc gcaaaatggc aaatggcacg tgaagctgtc
      181 gatattgggg aactgtggtg gttggcaaat gactaattaa gttagtcaag gcgccatcct
      241 catgaaaact gtgtaacata ataaccgaag tgtcgaaaag gtggcacctt gtccaattga
      301 acacgctcga tgaaaaaaat aagatatata taaggttaag taaagcgtct gttagaaagg
      361 aagtttttcc tttttcttgc tctcttgtct tttcatctac tatttccttc gtgtaataca
      421 gggtcgtcag atacatagat acaattctat tacccccatc catacaatgc catctcattt
      481 cgatactgtt caactacacg ccggtcaaga gaaccctggt gacaatgctc acagatccag
      541 agctgtacca atttacgcca ccacttctta tgttttcgaa aactctaagc atggttcgca
      601 attgtttggt ttagaagttc caggttacgt ctattcccgt ttccaaaacc caaccagtaa
      661 tgttttggaa gaaagaattg ctgctttaga aggtggtgct gctgctttgg ctgtttcctc
      721 cggtcaagcc gctcaaaccc ttgccatcca aggtttggca cacactggtg acaacatcgt
      781 ttccacttct tacttatacg gtggtactta taaccagttc aaaatctcgt tcaaaagatt
      841 tggtatcgag gctagatttg ttgaaggtga caatccagaa gaatttgaaa aggtctttga
      901 tgaaagaacc aaggctgttt atttggaaac cattggtaat ccaaagtaca atgttccgga
      961 ttttgaaaaa attgttgcaa ttgctcacaa acacggtatt ccagttgtcg ttgacaacac
     1021 atttggtgcc ggtggttact tctgtcagcc aattaaatac ggtgctgata ttgtaacaca
     1081 ttctgctacc aaatggattg gtggtcatgg tactactatc ggtggtatta ttgttgactc
     1141 tggtaagttc ccatggaagg actacccaga aaagttccct caattctctc aacctgccga
     1201 aggttatcac ggtactatct acaatgaagc ctacggtaac ttggcataca tcgttcatgt
     1261 tagaactgaa ctattaagag atttgggtcc attgatgaac ccatttgcct ctttcttgct
     1321 actacaaggt gttgaaacat tatctttgag agctgaaaga cacggtgaaa atgcattgaa
     1381 gttagccaaa tggttagaac aatccccata cgtatcttgg gtttcatacc ctggtttagc
     1441 atctcattct catcatgaaa atgctaagaa gtatctatct aacggtttcg gtggtgtctt
     1501 atctttcggt gtaaaagact taccaaatgc cgacaaggaa actgacccat tcaaactttc
     1561 tggtgctcaa gttgttgaca atttaaagct agcctctaac ttggccaatg ttggtgatgc
     1621 caagacctta gtcattgctc catacttcac tacccacaaa caattaaatg acaaagaaaa
     1681 gttggcatct ggtgttacca aggacttaat tcgtgtctct gttggtatcg aatttattga
     1741 tgacattatt gcagacttcc agcaatcttt tgaaactgtt ttcgctggcc aaaaaccatg
     1801 agtgtgcgta atgagttgta aaattatgta taaacctact ttctctcaca agtactatac
     1861 ttttataaaa cgaactttat tgaaatgaat atcctttttt tcccttgtta catgtcgtga
     1921 ctcgtacttt gaacctaaat tgttctaaca tcaaagaaca gtgttaattc gcagtcgaga
     1981 agaaaaatat ggtgaacaag actcatctac ttcatgagac tactttacgc ctcctataaa
     2041 gctgtcacac tggataaatt tattgtagga ccaagttaca aaagaggatg atggaggttt
     2101 ctttacaata aagaagcaca tgtgtgttaa cgtttttagt atttgcttgt tatgtaaatc
     2161 aggaaaactt cgcgggattt ggttggatgc tactttccat acaataaata ttatagatca
     2221 aaaaagccaa attacaagta aagattagta aagctgttgc aattccatcg ttgataaaaa
     2281 tgttagttat taaatataaa agtcagaata ggtgaacttg gatttaattg ttggcatttc
     2341 gttgctgcta gaggccataa tattagatag ccaggacata ctaggtctcc tcgtggtata
     2401 ggaatccata aaatggaatt ggtgacgatc ggatgtgctg caaggcgatt aagttgggta
     2461 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac
     2521 tcactatagg gcgaattgga gaaacagaat aatatataaa cctgtataat ataaccttga
     2581 agactatatt tcttttcgcg gccgcctata gggcgaattg gagctccacc gcggtggctc
     2641 tagaactagt ggatcccccg ggctgcagga attcgatatc aagcttatcg ataccgtcga
     2701 cctcgagggg gggcccggta cccagcttcg gccgtattta cagataacga tgtcgttatt
     2761 atatatatat atatatattg tcacttgtac ccagcttttg ttccctttag tgagggttaa
     2821 tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     2881 caacagctgt cagatatagt cggattgccc ttttaagcaa tcaatagtgt tttatttgca
     2941 acaatgtcgt catagtttaa tatgtcctat aagatgttaa cttgctcaac attcaacaaa
     3001 gtttggtctc ttggcccagt tggttaaggc accgtgctaa taacgcgggg atcagcggtt
     3061 cgatcccgct agagaccatt tatttttaaa ttcgaccgtc tgacagacgg tgcacttatt
     3121 acttaatttt tttttttttt taatcctcgt taaactgagt acataatcag caacaatata
     3181 atataaaagt tttgcttttt tatgtacgta caataggggt acttaaataa ataaataaac
     3241 aataaaacaa tcaataaata aataatatac gatcataaat tacgagatat atttcatagt
     3301 cttgacttct ccagccgaac gtcattaaat ttaaggcaga aaagataaac aagggcgcgc
     3361 cagagcctgc aggcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3421 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3481 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3541 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3601 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3661 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3721 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3781 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3841 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3901 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3961 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4021 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4081 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4141 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4201 tcctttgatc ttttgtttaa acctacgggg tctgacgctc agtggaacga aaactcacgt
     4261 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4321 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4381 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4441 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4501 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4561 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4621 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4681 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4741 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4801 tccttcggtc ctccgatggt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4861 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4921 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4981 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5041 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5101 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5161 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5221 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5281 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5341 acatttcccc gaaaagtgcc acct
//
