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pPpT4_MFalpha_eGFP vector (V017490)

Basic Vector Information

Vector Name:
pPpT4_MFalpha_eGFP
Antibiotic Resistance:
Bleomycin
Length:
4477 bp
Type:
Yeast Expression
Replication origin:
ori
Host:
Yeast
Copy Number:
High copy number
Promoter:
AOX1
Growth Strain(s):
DH5alpha
Growth Temperature:
37℃

pPpT4_MFalpha_eGFP vector Vector Map

pPpT4_MFalpha_eGFP4477 bp600120018002400300036004200P_AOX1_Syn_dBamHIAOX1 promoterrv(pPpT4)Signal sequence (MF-alpha)Gene of Interest GOI (eGFP)fw(pPpT4)AOX1 terminatorP_ILV5P_EM72_Synzeocin resistance proteinAOD terminatorori

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

pPpT4_MFalpha_eGFP vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                4477 bp DNA     circular SYN 29-MAY-2024
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4477)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4477)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Imported using the Genbank importer. File name: 
            pPpT4_Alph_formatiert.gb
FEATURES             Location/Qualifiers
     source          1..4477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          140..1070
                     /mol_type="other DNA"
                     /db_xref="taxon:1182041"
                     /organism="Escherichia coli-Pichia pastoris shuttle vector
                     pPpT4"
     source          2972..3346
                     /mol_type="other DNA"
                     /db_xref="taxon:1182041"
                     /organism="Escherichia coli-Pichia pastoris shuttle vector
                     pPpT4"
     source          3348..3823
                     /mol_type="other DNA"
                     /db_xref="taxon:1182041"
                     /organism="Escherichia coli-Pichia pastoris shuttle vector
                     pPpT4"
     misc_feature    6..131
                     /label=P_AOX1_Syn_dBamHI
     regulatory      140..1070
                     /label=AOX1 promoter
                     /note="AOX1 promoter"
                     /regulatory_class="promoter"
     primer_bind     complement(1071..1083)
                     /label=rv(pPpT4)
     CDS             1084..1350
                     /codon_start=1
                     /label=Signal sequence (MF-alpha)
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
     CDS             1351..2067
                     /codon_start=1
                     /label=Gene of Interest GOI (eGFP)
                     /translation="ASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     2068..2087
                     /label=fw(pPpT4)
     terminator      2088..2334
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2349..2906
                     /label=P_ILV5
     promoter        2907..2970
                     /label=P_EM72_Syn
     CDS             2972..3346
                     /codon_start=1
                     /product="zeocin resistance protein"
                     /label=zeocin resistance protein
                     /protein_id="AFJ20653.1"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     regulatory      3348..3823
                     /label=AOD terminator
                     /note="AOD terminator"
                     /regulatory_class="terminator"
     rep_origin      3870..4458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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