pYM33- EBFP Plasmid

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  • Model: PVT4015
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pYM33-EBFP

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pYM33-EBFP,Plasmid pYM33-EBFP,pYM33-EBFP vector

pYM33-EBFP Plasmid Information

Promoter: SP6, TEF, T7

Replicon: ori

Terminator: ADH1

Plasmid classification: yeast series, Pichia pastoris expression vector

Plasmid size: 4946bp

Plasmid label: EBFP

Prokaryotic resistance: Amp

Cloned strain: DH5 alpha

Culture conditions: 37 centigrade, aerobic LB

Expression host: yeast cells

5'sequencing primers: T7:TAATACGACTCACTATAGGG

3'sequencing primers: primers designed according to sequence

Use: Yeast expression

pYM33-EBFP Plasmid Description

pYM33-EBFP plasmid

pYM33-EBFP Plasmid Sequence

 

LOCUS       Exported                4946 bp ds-DNA     circular SYN 19-SEP-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4946)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, September 19, 2016 from SnapGene Viewer 3.2.1
            
FEATURES             Location/Qualifiers
     source          1..4946
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             79..795
                     /codon_start=1
                     /product="enhanced blue variant of GFP"
                     /note="EBFP"
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             802..828
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /note="HA"
                     /translation="YPYDVPDYA"
     terminator      861..1048
                     /gene="S. cerevisiae ADH1"
                     /note="ADH1 terminator"
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     gene            1123..2479
                     /note="kanMX"
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1123..1466
                     /note="TEF promoter"
                     /note="Ashbya gossypii TEF promoter"
     CDS             1467..2276
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      2282..2479
                     /note="TEF terminator"
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(2584..2602)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2860..3448)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3619..4479)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4480..4584)
                     /gene="bla"
                     /note="AmpR promoter"
     promoter        4930..2
                     /note="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatccgg agcaggtgct
       61 ggtgctggtg ctggagcaat gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      121 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      181 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
      241 gtgccctggc ccaccctcgt gaccaccctg acccacggcg tgcagtgctt cagccgctac
      301 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
      361 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
      421 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
      481 aacatcctgg ggcacaagct ggagtacaac ttcaacagcc acaacgtcta tatcatggcc
      541 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
      601 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
      661 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
      721 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
      781 gagctgtaca agtaaggatc ctatccatat gacgttccag attacgctgc tcagtgctag
      841 ggcgcgccac ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag
      901 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
      961 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg
     1021 tcgctcttat tgaccacacc tctaccggca gatccgctag ggataacagg gtaatataga
     1081 tctgtttagc ttgcctcgtc cccgccgggt cacccggcca gcgacatgga ggcccagaat
     1141 accctccttg acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac
     1201 atttagccca tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg
     1261 gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc
     1321 acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat
     1381 ttgccactga ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct
     1441 cacatcacat ccgaacataa acaaccatgg gtaaggaaaa gactcacgtt tcgaggccgc
     1501 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     1561 ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca gagttgtttc
     1621 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     1681 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     1741 catggttact caccactgcg atccccggca aaacagcatt ccaggtatta gaagaatatc
     1801 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     1861 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     1921 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     1981 ctgttgaaca agtctggaaa gaaatgcata agcttttgcc attctcaccg gattcagtcg
     2041 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
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