LOCUS       sequence-108542-        3833 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 3833)
  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 3833)
  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: pRG203MX
FEATURES             Location/Qualifiers
     source          1..3833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             273..923
                     /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     1262..1284
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1276..1293
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1277..1293
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1300..1319
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1300..1318
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1427..1443
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1427..1443
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1477..1493)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1478..1494)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(1592..1610)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1631..1647)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1631..1647)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1644..1666)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1655..1671
                     /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(1905..1922)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2076..2664)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2156..2175)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2843..3634,3635..3703))
                     /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     3466..3485
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3704..3808)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgaaa gaacgcggca ttagcaacat ttgcgaagcg aacgattttg cgaactgcta
       61 ttaattaact ctggcgcgcc tcctctagta cactctatat ttttttatgc ctcggtaatg
      121 attttcattt ttttttttcc acctagcgga tgactctttt tttttcttag cgattggcat
      181 tatcacataa tgaattatac attatataaa gtaatgtgat ttcttcgaag aatatactaa
      241 aaaatgagca ggcaagataa acgaaggcac atatgaggag ggcttttgta gaaagaaata
      301 cgaacgaaac gaaaatcagc gttgccatcg ctttggacaa agctccctta cctgaagagt
      361 cgaattttat tgatgaactt ataacttcca agcatgcaaa ccaaaaggga gaacaagtaa
      421 tccaagtaga cacgggaatt ggattcttgg atcacatgta tcatgcactg gctaaacatg
      481 caggctggag cttacgactt tactcaagag gtgatttaat catcgatgat catcacactg
      541 cagaagatac tgctattgca cttggtattg cattcaagca ggctatgggt aactttgccg
      601 gtgttaaaag atttggacat gcttattgtc cacttgacga agctctttcc agaagcgtag
      661 ttgacttgtc gggacggccc tatgctgtta tcgatttggg attaaagcgt gaaaaggttg
      721 gggaattgtc ctgtgaaatg atccctcact tactatattc cttttcggta gctgctggaa
      781 ttactttgca tgttacctgc ttatatggta gtaatgacca tcatcgtgct gaaagcgctt
      841 ttaaatctct ggctgttgcc atgcgcgcgg ctacttctct tactggaagt tctgaagtcc
      901 caagcacgaa gggagtgttg taaagatgaa ttggattatg tcaggaaaag aacgacaatt
      961 ttgcatccaa attgtctaaa ttttagagtt gcttgaaaac aatagaacct tacttgcttt
     1021 ataattacgt taattagaag cgttatctcg tgaaggaata tagtacgtag ccgtataaat
     1081 tgaattgaat gttcagctta tagaatagag acactttgct gttcaatgcg tcgtcactta
     1141 ccatactcac tttattatac gactttaagt ataaactccg cggttatggt aaaattaatg
     1201 atgcacaaac gtccgatcgg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     1261 tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc
     1321 gaattggaga aacagaataa tatataaacc tgtataatat aaccttgaag actatatttc
     1381 ttttcgcggc cgcctatagg gcgaattgga gctccaccgc ggtggctcta gaactagtgg
     1441 atcccccggg ctgcaggaat tcgatatcaa gcttatcgat accgtcgacc tcgagggggg
     1501 gcccggtacc cagcttcggc cgtatttaca gataacgatg tcgttattat atatatatat
     1561 atatattgtc acttgtaccc agcttttgtt ccctttagtg agggttaatt tcgagcttgg
     1621 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca acagctgaca
     1681 ccgattattt aaagctgcag catacgatat atatacatgt gtatatatgt atacctatga
     1741 atgtcagtaa gtatgtatac gaacagtatg atactgaaga tgacaaggta atgcatcatt
     1801 ctatacgtgt cattctgaac gaggcgcgcc tctccctgca ggcattaatg aatcggccaa
     1861 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     1921 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1981 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2041 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2101 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2161 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2221 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2281 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2341 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2401 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2461 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2521 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2581 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2641 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttgtttaaa cctacggggt
     2701 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2761 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2821 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2881 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     2941 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3001 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3061 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3121 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3181 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3241 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatggtt gtcagaagta
     3301 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3361 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     3421 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3481 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3541 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3601 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3661 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3721 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     3781 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cct
//
