LOCUS       sequence-108544-        5125 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 5125)
  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 5125)
  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: pRG223MX
FEATURES             Location/Qualifiers
     source          1..5125
                     /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             1565..2215
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQK
                     GEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQA
                     MGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLYS
                     FSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     primer_bind     2554..2576
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2568..2585
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2569..2585
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2592..2611
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2592..2610
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2719..2735
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2719..2735
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2769..2785)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2770..2786)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(2884..2902)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2923..2939)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2923..2939)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2936..2958)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2947..2963
                     /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(3197..3214)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3368..3956)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3448..3467)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4135..4926,4927..4995))
                     /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     4758..4777
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4996..5100)
                     /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 cctcctctag
     1381 tacactctat atttttttat gcctcggtaa tgattttcat tttttttttt ccacctagcg
     1441 gatgactctt tttttttctt agcgattggc attatcacat aatgaattat acattatata
     1501 aagtaatgtg atttcttcga agaatatact aaaaaatgag caggcaagat aaacgaaggc
     1561 acatatgagg agggcttttg tagaaagaaa tacgaacgaa acgaaaatca gcgttgccat
     1621 cgctttggac aaagctccct tacctgaaga gtcgaatttt attgatgaac ttataacttc
     1681 caagcatgca aaccaaaagg gagaacaagt aatccaagta gacacgggaa ttggattctt
     1741 ggatcacatg tatcatgcac tggctaaaca tgcaggctgg agcttacgac tttactcaag
     1801 aggtgattta atcatcgatg atcatcacac tgcagaagat actgctattg cacttggtat
     1861 tgcattcaag caggctatgg gtaactttgc cggtgttaaa agatttggac atgcttattg
     1921 tccacttgac gaagctcttt ccagaagcgt agttgacttg tcgggacggc cctatgctgt
     1981 tatcgatttg ggattaaagc gtgaaaaggt tggggaattg tcctgtgaaa tgatccctca
     2041 cttactatat tccttttcgg tagctgctgg aattactttg catgttacct gcttatatgg
     2101 tagtaatgac catcatcgtg ctgaaagcgc ttttaaatct ctggctgttg ccatgcgcgc
     2161 ggctacttct cttactggaa gttctgaagt cccaagcacg aagggagtgt tgtaaagatg
     2221 aattggatta tgtcaggaaa agaacgacaa ttttgcatcc aaattgtcta aattttagag
     2281 ttgcttgaaa acaatagaac cttacttgct ttataattac gttaattaga agcgttatct
     2341 cgtgaaggaa tatagtacgt agccgtataa attgaattga atgttcagct tatagaatag
     2401 agacactttg ctgttcaatg cgtcgtcact taccatactc actttattat acgactttaa
     2461 gtataaactc cgcggttatg gtaaaattaa tgatgcacaa acgtccgatc ggatgtgctg
     2521 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     2581 ccagtgaatt gtaatacgac tcactatagg gcgaattgga gaaacagaat aatatataaa
     2641 cctgtataat ataaccttga agactatatt tcttttcgcg gccgcctata gggcgaattg
     2701 gagctccacc gcggtggctc tagaactagt ggatcccccg ggctgcagga attcgatatc
     2761 aagcttatcg ataccgtcga cctcgagggg gggcccggta cccagcttcg gccgtattta
     2821 cagataacga tgtcgttatt atatatatat atatatattg tcacttgtac ccagcttttg
     2881 ttccctttag tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt
     2941 gtgaaattgt tatccgctca caacagctga caccgattat ttaaagctgc agcatacgat
     3001 atatatacat gtgtatatat gtatacctat gaatgtcagt aagtatgtat acgaacagta
     3061 tgatactgaa gatgacaagg taatgcatca ttctatacgt gtcattctga acgaggcgcg
     3121 cctctccctg caggcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3181 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3241 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3301 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3361 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3421 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3481 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3541 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3601 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3661 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3721 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3781 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3841 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3901 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3961 atcctttgat cttttgttta aacctacggg gtctgacgct cagtggaacg aaaactcacg
     4021 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4081 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4141 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4201 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4261 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4321 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4381 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4441 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4501 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4561 ctccttcggt cctccgatgg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4621 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4681 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4741 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4801 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4861 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4921 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4981 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5041 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5101 cacatttccc cgaaaagtgc cacct
//
