LOCUS       Exported                3619 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pBA2219
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3619)
  AUTHORS   Austin BP, Waugh DS
  TITLE     Isolation of Metarhizium anisopliae carboxypeptidase A with native 
            disulfide bonds from the cytosol of Escherichia coli BL21(DE3).
  JOURNAL   Protein Expr Purif. 2012 Mar;82(1):116-24. Epub 2011 Dec 14.
  PUBMED    22197595
REFERENCE   2  (bases 1 to 3619)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Expr Purif."; date: "2012-03"; volume: "82(1)"; pages: "116-24. Epub
            2011 Dec 14"
FEATURES             Location/Qualifiers
     source          1..3619
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      108..653
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     554..573
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     770..787
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1237..1256)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1248..2063
                     /codon_start=1
                     /gene="aph(3)-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1321..1340)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2665..2684
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2665..2683
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2715..2737
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2744..2776
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     protein_bind    2786..2810
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3460..3484)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3533..3551)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3547..3594
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"