LOCUS       Exported                4610 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBIOTRX-BirA, complete sequence.
ACCESSION   AF044308
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4610)
  AUTHORS   LaVallie ER, DiBlasio EA, Kovacic S, Grant KL, Schendel PF, McCoy 
            JM.
  TITLE     A thioredoxin gene fusion expression system that circumvents 
            inclusion body formation in the E. coli cytoplasm
  JOURNAL   Biotechnology (N.Y.) 11 (2), 187-193 (1993)
  PUBMED    7763371
REFERENCE   2  (bases 1 to 4610)
  AUTHORS   Smith PA, Tripp BC, DiBlasio-Smith EA, Lu Z, LaVallie ER, McCoy JM.
  TITLE     A plasmid expression system for quantitative in vivo biotinylation 
            of thioredoxin fusion proteins in Escherichia coli
  JOURNAL   Nucleic Acids Res. 26 (6), 1414-1420 (1998)
  PUBMED    9490786
REFERENCE   3  (bases 1 to 4610)
  AUTHORS   Smith PA, Tripp BC, DiBlasio-Smith EA, Lu Z, LaVallie ER, McCoy JM.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-JAN-1998) Molecular Biology and Gene Expression, 
            Genetics Institute, Inc., 87 Cambridge Park Drive, Cambridge, MA 
            02140, USA
REFERENCE   4  (bases 1 to 4610)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4610
                     /organism="Cloning vector pBIOTRX-BirA"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:72145"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             201..1061
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AAC40209.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            201..1061
                     /gene="bla"
                     /label=bla
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      2159..2187
                     /regulatory_class="promoter"
                     /note="lambda pL"
     misc_feature    2724..2792
                     /note="Tag 81; biotinylation recognition sequence"
     CDS             2793..3122
                     /codon_start=1
                     /gene="trxA"
                     /product="Eschericia coli thioredoxin"
                     /label=trxA
                     /protein_id="AAC40210.1"
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     gene            2793..3122
                     /gene="trxA"
                     /label=trxA
     CDS             2793..3119
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             3154..4119
                     /codon_start=1
                     /gene="birA"
                     /product="Eschericia coli biotin-holoenzyme synthetase"
                     /label=birA
                     /protein_id="AAC40211.1"
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     gene            3154..4119
                     /gene="birA"
                     /label=birA
     regulatory      4138..4204
                     /regulatory_class="terminator"
                     /note="AspA gene transcription terminator"
     primer_bind     complement(4214..4230)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"