LOCUS       sequence-61112-e        4732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with mCherry with SpHis5 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4732)
  TITLE     Yeast tagging plasmids.
REFERENCE   2  (bases 1 to 4732)
  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"
COMMENT     Sequence Label: pFA6a-link-mCherry-SpHis5
FEATURES             Location/Qualifiers
     source          1..4732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             5..715
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(150..168)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     418..437
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      736..923
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(791..815)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            998..2198
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        998..1341
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1342..1995
                     /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      2001..2198
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2302..2321)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2303..2321)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2408..2425)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2579..3167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2659..2678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3338..4129,4130..4198))
                     /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     3961..3980
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4199..4303)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4371..4389
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4427..4449)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4549..4568
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4649..4667
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4649..4666
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 taacatggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
       61 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
      121 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
      181 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
      241 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
      301 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
      361 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
      421 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
      481 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
      541 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
      601 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
      661 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtaaggcgc
      721 gccacttcta aataagcgaa tttcttatga tttatgattt ttattattaa ataagttata
      781 aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg aaaattctta
      841 ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagta tgaggtcgct
      901 cttattgacc acacctctac cggcagatcc gctagggata acagggtaat atagatctgt
      961 ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac atggaggccc agaataccct
     1021 ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta
     1081 gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg
     1141 aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga
     1201 cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc
     1261 actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat
     1321 cacatccgaa cataaacaac catgggtagg agggcttttg tagaaagaaa tacgaacgaa
     1381 acgaaaatca gcgttgccat cgctttggac aaagctccct tacctgaaga gtcgaatttt
     1441 attgatgaac ttataacttc caagcatgca aaccaaaagg gagaacaagt aatccaagta
     1501 gacacgggaa ttggattctt ggatcacatg tatcatgcac tggctaaaca tgcaggctgg
     1561 agcttacgac tttactcaag aggtgattta atcatcgatg atcatcacac tgcagaagat
     1621 actgctattg cacttggtat tgcattcaag caggctatgg gtaactttgc cggcgttaaa
     1681 agatttggac atgcttattg tccacttgac gaagctcttt ctagaagcgt agttgacttg
     1741 tcgggacggc cctatgctgt tatcgatttg ggattaaagc gtgaaaaggt tggggaattg
     1801 tcctgtgaaa tgatccctca cttactatat tccttttcgg tagcagctgg aattactttg
     1861 catgttacct gcttatatgg tagtaatgac catcatcgtg ctgaaagcgc ttttaaatct
     1921 ctggctgttg ccatgcgcgc ggctactagt cttactggaa gttctgaagt cccaagcacg
     1981 aagggagtgt tgtaaagagt actgacaata aaaagattct tgttttcaag aacttgtcat
     2041 ttgtatagtt tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata
     2101 ttttttttcg cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat
     2161 catgcgtcaa tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc
     2221 gccatccagt ttaaacgagc tcgaattcat cgatgatatc agatccacta gtggcctatg
     2281 cggccgcgga tctgccggtc tccctatagt gagtcgtatt aatttcgata agccaggtta
     2341 acctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2401 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2461 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2521 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2581 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2641 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2701 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2761 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2821 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2881 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2941 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3001 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3061 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3121 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3181 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3241 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3301 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3361 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3421 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3481 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3541 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3601 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3661 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3721 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3781 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3841 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3901 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3961 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4021 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4081 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4141 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4201 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4261 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4321 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     4381 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     4441 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     4501 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     4561 agattgtact gagagtgcac catatggaca tattgtcgtt agaacgcggc tacaattaat
     4621 acataacctt atgtatcata cacatacgat ttaggtgaca ctatagaacg cggccgccag
     4681 ctgaagcttc gtacgctgca ggtcgacgga tcggtgacgg tgctggttta at
//
