LOCUS       sequence-422261-        4137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid backbone for yeast expression, use auxotrophic marker HIS5 
            (Schizosaccharomyces Pombe).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4137)
  AUTHORS   He CW, Cui XF, Ma SJ, Xu Q, Ran YP, Chen WZ, Mu JX, Li H, Zhu J, 
            Gong Q, Xie Z
  TITLE     Membrane recruitment of Atg8 by Hfl1 facilitates turnover of 
            vacuolar membrane proteins in yeast cells approaching stationary 
            phase.
  JOURNAL   BMC Biol. 2021 Jun 4;19(1):117. doi: 10.1186/s12915-021-01048-7.
  PUBMED    34088313
REFERENCE   2  (bases 1 to 4137)
  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: "BMC Biol.";
            date: "2021-06-4"; volume: "19(1):117. doi"; pages: " 
            10.1186/s12915-021-01048-7"
COMMENT     Sequence Label: ClhN-HIS5
FEATURES             Location/Qualifiers
     source          1..4137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      101..288
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(156..180)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    340..373
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            428..1628
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        428..771
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             772..1425
                     /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      1431..1628
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    1688..1721
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(1774..1793)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1775..1793)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1880..1897)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2051..2639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2131..2150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2810..3601,3602..3670))
                     /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     3433..3452
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3671..3775)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3843..3861
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3899..3921)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4021..4040
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4121..4137,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4121..4137,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg tgggattcct gggttaatta
       61 acatcgctag ccctaggtga ggcgcgccac ttctaaataa gcgaatttct tatgatttat
      121 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
      181 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
      241 ttctcaggta tagtatgagg tcgctcttat tgaccacacc tctaccggca gatccgctag
      301 ggataacagg gtaatatctg caggtcgaca acccttaata taacttcgta taatgtatgc
      361 tatacgaagt tattaggtct agagatctgt ttagcttgcc tcgtccccgc cgggtcaccc
      421 ggccagcgac atggaggccc agaataccct ccttgacagt cttgacgtgc gcagctcagg
      481 ggcatgatgt gactgtcgcc cgtacattta gcccatacat ccccatgtat aatcatttgc
      541 atccatacat tttgatggcc gcacggcgcg aagcaaaaat tacggctcct cgctgcagac
      601 ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct
      661 gtagagaaat ataaaaggtt aggatttgcc actgaggttc ttctttcata tacttccttt
      721 taaaatcttg ctaggataca gttctcacat cacatccgaa cataaacaac catgggtagg
      781 agggcttttg tagaaagaaa tacgaacgaa acgaaaatca gcgttgccat cgctttggac
      841 aaagctccct tacctgaaga gtcgaatttt attgatgaac ttataacttc caagcatgca
      901 aaccaaaagg gagaacaagt aatccaagta gacacgggaa ttggattcct ggatcacatg
      961 tatcatgcac tggctaaaca tgcaggctgg agcttacgac tttactcaag aggtgattta
     1021 atcatcgatg atcatcacac tgcagaagat actgctattg cacttggtat tgcattcaag
     1081 caggctatgg gtaactttgc cggcgttaaa agatttggac atgcttattg tccacttgac
     1141 gaagcccttt ctagaagcgt agttgacttg tcgggacggc cctatgctgt tatcgatttg
     1201 ggattaaagc gtgaaaaggt tggggaattg tcctgtgaaa tgatccctca cttactatat
     1261 tccttttcgg tagcagctgg aattactttg catgttacct gcttatatgg tagtaatgac
     1321 catcatcgtg ctgaaagcgc ttttaaatct ctggctgttg ccatgcgcgc ggctactagt
     1381 cttactggaa gttctgaagt cccaagcacg aagggagtgt tgtaaagagt actgacaata
     1441 aaaagattct tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc
     1501 tattttaatc aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag
     1561 atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg
     1621 ctatactgct gtcgattcga tactaacgcc gccatccagt ttaaacgagc tctcgagaac
     1681 ccttaatata acttcgtata atgtatgcta tacgaagtta ttaggtgata tcagatccac
     1741 tagtggccta tgcggccgcg gatctgccgg tctccctata gtgagtcgta ttaatttcga
     1801 taagccaggt taacctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     1861 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     1921 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     1981 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2041 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2101 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2161 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2221 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2281 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2341 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2401 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2461 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2521 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2581 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2641 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2701 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2761 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2821 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     2881 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     2941 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3001 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3061 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3121 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3181 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3241 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3301 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3361 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3421 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3481 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3541 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3601 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3661 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3721 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3781 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     3841 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     3901 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     3961 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     4021 cggcatcaga gcagattgta ctgagagtgc accatatgga catattgtcg ttagaacgcg
     4081 gctacaatta atacataacc ttatgtatca tacacatacg atttaggtga cactata
//
