LOCUS       sequence-318460-        5093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for C-terminal BirA* tagging of proteins in Saccharomyces 
            cerevisiae; Selection marker: HIS3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5093)
  AUTHORS   Salvatori R, Kehrein K, Singh AP, Aftab W, Moller-Hergt BV, Forne I,
            Imhof A, Ott M
  TITLE     Molecular Wiring of a Mitochondrial Translational Feedback Loop.
  JOURNAL   Mol Cell. 2020 Feb 20;77(4):887-900.e5. doi: 
            10.1016/j.molcel.2019.11.019. Epub 2019 Dec 26.
  PUBMED    31883951
REFERENCE   2  (bases 1 to 5093)
  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"; doi: 
            "10.1016/j.molcel.2019.11.019"; journalName: "Mol Cell"; date: 
            "2020-02-20- 20"; volume: "77"; issue: "4"; pages: "887-900.e5"
COMMENT     Sequence Label: pYM28-BirA*
FEATURES             Location/Qualifiers
     source          1..5093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..27
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      60..247
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(115..139)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            322..1522
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        322..665
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             666..1319
                     /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      1325..1522
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1626..1645)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1627..1645)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1732..1749)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1903..2491)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1983..2002)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2662..3453,3454..3522))
                     /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     3285..3304
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3523..3627)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3695..3713
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3751..3773)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3873..3892
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3973..3991
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3973..3990
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4032..4055
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 tatccatatg acgttccaga ttacgctgct cagtgctagg gcgcgccact tctaaataag
       61 cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat
      121 acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt
      181 tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct
      241 ctaccggcag atccgctagg gataacaggg taatatagat ctgtttagct tgcctcgtcc
      301 ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac
      361 gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat
      421 gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc
      481 tcctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc
      541 cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt
      601 catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa
      661 caaccatggg taggagggct tttgtagaaa gaaatacgaa cgaaacgaaa atcagcgttg
      721 ccatcgcttt ggacaaagct cccttacctg aagagtcgaa ttttattgat gaacttataa
      781 cttccaagca tgcaaaccaa aagggagaac aagtaatcca agtagacacg ggaattggat
      841 tcctggatca catgtatcat gcactggcta aacatgcagg ctggagctta cgactttact
      901 caagaggtga tttaatcatc gatgatcatc acactgcaga agatactgct attgcacttg
      961 gtattgcatt caagcaggct atgggtaact ttgccggcgt taaaagattt ggacatgctt
     1021 attgtccact tgacgaagcc ctttctagaa gcgtagttga cttgtcggga cggccctatg
     1081 ctgttatcga tttgggatta aagcgtgaaa aggttgggga attgtcctgt gaaatgatcc
     1141 ctcacttact atattccttt tcggtagcag ctggaattac tttgcatgtt acctgcttat
     1201 atggtagtaa tgaccatcat cgtgctgaaa gcgcttttaa atctctggct gttgccatgc
     1261 gcgcggctac tagtcttact ggaagttctg aagtcccaag cacgaaggga gtgttgtaaa
     1321 gagtactgac aataaaaaga ttcttgtttt caagaacttg tcatttgtat agttttttta
     1381 tattgtagtt gttctatttt aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga
     1441 catcatctgc ccagatgcga agttaagtgc gcagaaagta atatcatgcg tcaatcgtat
     1501 gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa cgccgccatc cagtttaaac
     1561 gagctcgaat tcatcgatga tatcagatcc actagtggcc tatgcggccg cggatctgcc
     1621 ggtctcccta tagtgagtcg tattaatttc gataagccag gttaacctgc attaatgaat
     1681 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1741 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1801 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1861 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1921 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1981 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2041 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2101 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2161 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2221 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2281 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2341 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2401 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2461 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2521 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2581 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2641 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2701 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2761 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     2821 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2881 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     2941 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3001 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3061 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3121 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3181 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3241 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3301 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3361 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3421 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3481 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3541 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3601 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     3661 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     3721 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     3781 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     3841 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     3901 gcaccatatg gacatattgt cgttagaacg cggctacaat taatacataa ccttatgtat
     3961 catacacata cgatttaggt gacactatag aacgcggccg ccagctgaag cttcgtacgc
     4021 tgcaggtcga cgattacaag gacgacgacg acaagacgcg tggtggtggt ggttctggtg
     4081 gtggtggttc tatgaaggac aacaccgtgc ccctgaagct gatcgccctg ctggctaacg
     4141 gcgagttcca ctctggcgag cagctgggcg agacactggg catgtctaga gccgccatca
     4201 acaagcacat ccagaccctg agagactggg gcgtggacgt gttcaccgtg cctggcaagg
     4261 gctacagcct gcccgaacct atccagctgc tgaacgccaa gcagatcctg ggccagctgg
     4321 acggcggatc tgtggctgtg ctgcctgtga tcgacagcac caaccagtac ctgctggaca
     4381 gaatcggcga gctgaagtcc ggcgacgcct gtatcgctga gtaccagcag gctggcagag
     4441 gcggcagggg cagaaagtgg ttctctccct tcggcgctaa cctgtacctg agcatgttct
     4501 ggcggctgga acagggacct gccgctgcta tcggactgag cctcgtgatc ggcatcgtga
     4561 tggccgaggt gctgagaaag ctgggcgccg acaaagtgcg cgtgaagtgg cccaacgacc
     4621 tgtatctgca ggaccggaag ctggccggca tcctggtgga actgacaggc aagacaggcg
     4681 acgccgccca gatcgtgatt ggcgctggca tcaacatggc tatgagaaga gtggaagagt
     4741 ccgtcgtgaa ccagggctgg atcaccctgc aggaagccgg catcaacctg gaccggaaca
     4801 cactggccgc catgctgatc agagagctga gagccgccct ggaactgttc gagcaggaag
     4861 gcctggcccc ctacctgtcc agatgggaga agctggacaa cttcatcaac agacccgtga
     4921 aactgatcat cggcgacaaa gaaatcctgg gcatcagcag aggcatcgac aagcagggcg
     4981 ctctgctgct ggaacaggac ggcatcatca agccctggat gggcggcgag atcagcctga
     5041 gatctgctga gaagggcggc agcggacctg gcggaggcgc tccttaagga tcc
//
