LOCUS       sequence-108541-        6656 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 6656)
  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 6656)
  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: pRG221
FEATURES             Location/Qualifiers
     source          1..6656
                     /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)."
     promoter        1408..1758
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     CDS             1759..3093
                     /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     3766..3788
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3780..3797
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3781..3797
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3804..3823
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3804..3822
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3931..3947
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3931..3947
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3981..3997)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3982..3998)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(4096..4114)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4135..4151)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4135..4151)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4148..4170)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4159..4175
                     /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(4728..4745)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4899..5487)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4979..4998)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5666..6457,6458..6526))
                     /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     6289..6308
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6527..6631)
                     /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 acacacacca taccttccca
     1381 tatgtcctgg cgcgccctta gctctcatta ttttttgctt tttctcttga ggtcacatga
     1441 tcgcaaaatg gcaaatggca cgtgaagctg tcgatattgg ggaactgtgg tggttggcaa
     1501 atgactaatt aagttagtca aggcgccatc ctcatgaaaa ctgtgtaaca taataaccga
     1561 agtgtcgaaa aggtggcacc ttgtccaatt gaacacgctc gatgaaaaaa ataagatata
     1621 tataaggtta agtaaagcgt ctgttagaaa ggaagttttt cctttttctt gctctcttgt
     1681 cttttcatct actatttcct tcgtgtaata cagggtcgtc agatacatag atacaattct
     1741 attaccccca tccatacaat gccatctcat ttcgatactg ttcaactaca cgccggtcaa
     1801 gagaaccctg gtgacaatgc tcacagatcc agagctgtac caatttacgc caccacttct
     1861 tatgttttcg aaaactctaa gcatggttcg caattgtttg gtttagaagt tccaggttac
     1921 gtctattccc gtttccaaaa cccaaccagt aatgttttgg aagaaagaat tgctgcttta
     1981 gaaggtggtg ctgctgcttt ggctgtttcc tccggtcaag ccgctcaaac ccttgccatc
     2041 caaggtttgg cacacactgg tgacaacatc gtttccactt cttacttata cggtggtact
     2101 tataaccagt tcaaaatctc gttcaaaaga tttggtatcg aggctagatt tgttgaaggt
     2161 gacaatccag aagaatttga aaaggtcttt gatgaaagaa ccaaggctgt ttatttggaa
     2221 accattggta atccaaagta caatgttccg gattttgaaa aaattgttgc aattgctcac
     2281 aaacacggta ttccagttgt cgttgacaac acatttggtg ccggtggtta cttctgtcag
     2341 ccaattaaat acggtgctga tattgtaaca cattctgcta ccaaatggat tggtggtcat
     2401 ggtactacta tcggtggtat tattgttgac tctggtaagt tcccatggaa ggactaccca
     2461 gaaaagttcc ctcaattctc tcaacctgcc gaaggttatc acggtactat ctacaatgaa
     2521 gcctacggta acttggcata catcgttcat gttagaactg aactattaag agatttgggt
     2581 ccattgatga acccatttgc ctctttcttg ctactacaag gtgttgaaac attatctttg
     2641 agagctgaaa gacacggtga aaatgcattg aagttagcca aatggttaga acaatcccca
     2701 tacgtatctt gggtttcata ccctggttta gcatctcatt ctcatcatga aaatgctaag
     2761 aagtatctat ctaacggttt cggtggtgtc ttatctttcg gtgtaaaaga cttaccaaat
     2821 gccgacaagg aaactgaccc attcaaactt tctggtgctc aagttgttga caatttaaag
     2881 ctagcctcta acttggccaa tgttggtgat gccaagacct tagtcattgc tccatacttc
     2941 actacccaca aacaattaaa tgacaaagaa aagttggcat ctggtgttac caaggactta
     3001 attcgtgtct ctgttggtat cgaatttatt gatgacatta ttgcagactt ccagcaatct
     3061 tttgaaactg ttttcgctgg ccaaaaacca tgagtgtgcg taatgagttg taaaattatg
     3121 tataaaccta ctttctctca caagtactat acttttataa aacgaacttt attgaaatga
     3181 atatcctttt tttcccttgt tacatgtcgt gactcgtact ttgaacctaa attgttctaa
     3241 catcaaagaa cagtgttaat tcgcagtcga gaagaaaaat atggtgaaca agactcatct
     3301 acttcatgag actactttac gcctcctata aagctgtcac actggataaa tttattgtag
     3361 gaccaagtta caaaagagga tgatggaggt ttctttacaa taaagaagca catgtgtgtt
     3421 aacgttttta gtatttgctt gttatgtaaa tcaggaaaac ttcgcgggat ttggttggat
     3481 gctactttcc atacaataaa tattatagat caaaaaagcc aaattacaag taaagattag
     3541 taaagctgtt gcaattccat cgttgataaa aatgttagtt attaaatata aaagtcagaa
     3601 taggtgaact tggatttaat tgttggcatt tcgttgctgc tagaggccat aatattagat
     3661 agccaggaca tactaggtct cctcgtggta taggaatcca taaaatggaa ttggtgacga
     3721 tcggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     3781 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg gagaaacaga
     3841 ataatatata aacctgtata atataacctt gaagactata tttcttttcg cggccgccta
     3901 tagggcgaat tggagctcca ccgcggtggc tctagaacta gtggatcccc cgggctgcag
     3961 gaattcgata tcaagcttat cgataccgtc gacctcgagg gggggcccgg tacccagctt
     4021 cggccgtatt tacagataac gatgtcgtta ttatatatat atatatatat tgtcacttgt
     4081 acccagcttt tgttcccttt agtgagggtt aatttcgagc ttggcgtaat catggtcata
     4141 gctgtttcct gtgtgaaatt gttatccgct cacaacagct gtcagatata gtcggattgc
     4201 ccttttaagc aatcaatagt gttttatttg caacaatgtc gtcatagttt aatatgtcct
     4261 ataagatgtt aacttgctca acattcaaca aagtttggtc tcttggccca gttggttaag
     4321 gcaccgtgct aataacgcgg ggatcagcgg ttcgatcccg ctagagacca tttattttta
     4381 aattcgaccg tctgacagac ggtgcactta ttacttaatt tttttttttt tttaatcctc
     4441 gttaaactga gtacataatc agcaacaata taatataaaa gttttgcttt tttatgtacg
     4501 tacaataggg gtacttaaat aaataaataa acaataaaac aatcaataaa taaataatat
     4561 acgatcataa attacgagat atatttcata gtcttgactt ctccagccga acgtcattaa
     4621 atttaaggca gaaaagataa acaagggcgc gccagagcct gcaggcatta atgaatcggc
     4681 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4741 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4801 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4861 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4921 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4981 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5041 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5101 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5161 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5221 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5281 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5341 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5401 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5461 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttgttt aaacctacgg
     5521 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5581 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5641 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5701 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5761 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5821 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5881 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5941 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6001 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6061 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatg gttgtcagaa
     6121 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6181 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6241 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6301 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6361 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6421 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6481 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6541 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6601 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacct
//
