pPpT4_MFalpha_eGFP vector (V017490)

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"