LOCUS       sequence-154309-        4293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gene Tagging vector HIS/RFP - Plasmid for monomeric Cherry (mCherry)
            labeling of endogenous proteins with a HIS selection marker, 
            originating from pSIVh (ID:81091).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4293)
  AUTHORS   Wosika V, Durandau E, Varidel C, Aymoz D, Schmitt M, Pelet S
  TITLE     New families of single integration vectors and gene tagging plasmids
            for genetic manipulations in budding yeast.
  JOURNAL   Mol Genet Genomics. 2016 Dec;291(6):2231-2240. Epub 2016 Sep 16.
  PUBMED    27637489
REFERENCE   2  (bases 1 to 4293)
  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: "Mol Genet 
            Genomics."; date: "2016-12"; volume: "291(6)"; pages: "2231-2240. 
            Epub 2016 Sep 16"
COMMENT     Sequence Label: GTH-r
FEATURES             Location/Qualifiers
     source          1..4293
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(88..151)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        178..196
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(358..376)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     626..645
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      971..1158
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1026..1050)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1197..1213)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1197..1213)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    1214..1247
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1749..2402
                     /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="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      2408..2605
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    2629..2662
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2663..2681
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2710..2728)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2761..3349)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2841..2860)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3520..4293,1..18,19..87))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4143..4162
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
       61 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
      121 tcagggttat tgtctcatga gcggatacat atttgaagta tcgttcgaat taattaaaat
      181 taaccctcac taaaggggca tgcgcggccg ctctagaact aaagggcgag gaggataaca
      241 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
      301 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
      361 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
      421 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
      481 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
      541 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
      601 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
      661 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
      721 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
      781 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
      841 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
      901 gcatggacga gctgtacaag ggatcttaaa tgctagcatt ttaactcgag ggcgcgccac
      961 ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa
     1021 aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt
     1081 gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg tcgctcttat
     1141 tgaccacacc tctaccggca gatccgctag ggataacagg gtaatataga tccatggtca
     1201 tagctgtttc ctgataactt cgtataatgt atgctatacg aagttatgtc tgtaggcgaa
     1261 cctaaccggc caattttcca agaccgctct gccgctgagc ttgcacccgt gccgctacac
     1321 aacacccaca catccgtaaa ccccccgcca tcggtatcac caaggttgtc tatgacgagg
     1381 cgtacaagag gacgtggggg ccgcctctcc aatgtcacca ataggaaaaa agaaagaccg
     1441 agtgcgagtt tctgacagaa gcctcaagaa ttttttttct ttcttcgact atgctggagg
     1501 tagagatgat cgaggcagta gtcaagtata tataaattaa ttagttccac caccttcttt
     1561 tcttgtgttg ctcctttcga tgctatttct ggctttttct aatttttttc atcgtttctt
     1621 actttcttcc taatatataa attcttttgc attttcattc ttcctctcca tttgttcatt
     1681 ctccttaaat tttttttagg tctcctttct ttcaagaaac acagtcatcg aactataatt
     1741 aactaaacat gggtaggagg gcttttgtag aaagaaatac gaacgaaacg aaaatcagcg
     1801 ttgctatcgc tttggacaaa gctcccttac ctgaagagtc gaattttatt gatgaactta
     1861 taacttccaa acatgcaaac caaaagggag aacaagtaat ccaagtagac acgggaattg
     1921 gattcttgga tcacatgtat catgcactgg ctaaacatgc aggctggagc ttacgacttt
     1981 actcaagagg tgatttaatc attgatgatc atcacacagc agaagatact gctattgcac
     2041 ttggtattgc attcaagcag gctatgggta actttgccgg cgttaaaaga tttggacatg
     2101 cttattgtcc acttgacgaa gctctttcaa gaagcgtagt tgatttgtcg ggacggccct
     2161 atgctgttat tgatttggga ttaaagcgtg aaaaggttgg ggaattgtcc tgtgaaatga
     2221 tccctcactt actatattcc ttttcggtag ctgctggaat tactttgcat gttacctgct
     2281 tatatggtag taatgaccat catcgtgctg aaagcgcttt taaatctctg gctgttgcca
     2341 tgcgcgcggc tacaagtctt actggaagtt ctgaagtccc aagcacgaag ggagtgttgt
     2401 aatcattact gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt
     2461 ttatattgta gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct
     2521 cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg
     2581 tatgtgaatg ctggtcgcta tactgctgtc gattcgatac taacgccgat aacttcgtat
     2641 aatgtatgct atacgaagtt attaatacga ctcactatag ggcgaattgg agctccaatt
     2701 cggacgtccc ctatagtgag tcgtattatt aattaattcg aaggccgcgt tgctggcgtt
     2761 tttccatagg ctcggccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2821 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2881 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2941 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3001 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3061 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3121 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3181 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3241 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3301 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3361 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3421 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3481 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3541 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tgcccgtcgt
     3601 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3661 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3721 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3781 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3841 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3901 aaggcgagtt acatgatccc ccatgttgtg aaaaaaagcg gttagctcct tcggtcctcc
     3961 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4021 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4081 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4141 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4201 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4261 tgcacccaac tgatcttcag catcttttac ttt
//
