LOCUS       Exported                4482 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Co-expression vector pCDF-BIRA, complete sequence.
ACCESSION   JF914075
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4482)
  AUTHORS   Keates T, Cooper CD, Savitsky P, Allerston CK, Phillips C, 
            Hammarstrom M, Daga N, Berridge G, Mahajan P, Burgess-Brown NA, 
            Muller S, Graslund S, Gileadi O.
  TITLE     Expressing the human proteome for affinity proteomics: optimising 
            expression of soluble protein domains and in vivo biotinylation
  JOURNAL   N Biotechnol 29 (5), 515-525 (2012)
  PUBMED    22027370
REFERENCE   2  (bases 1 to 4482)
  AUTHORS   Savitsky P, Gileadi O.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-MAY-2011) Structural Genomics Consortium, University 
            of Oxford, Old Road Campus Research Building, Roosevelt Drive, 
            Oxford, Oxfordshire OX3 7DQ, UK
REFERENCE   3  (bases 1 to 4482)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4482
                     /organism="Co-expression vector pCDF-BIRA"
                     /mol_type="other DNA"
                     /label=derived from pCDFDuet-1
                     /note="derived from pCDFDuet-1"
                     /db_xref="taxon:1088859"
     promoter        84..102
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      84..100
                     /regulatory_class="promoter"
                     /note="T7"
     protein_bind    103..127
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     misc_feature    105..125
                     /label=lac operator
                     /note="lac operator"
     CDS             171..1139
                     /codon_start=1
                     /gene="birA"
                     /product="BirA"
                     /label=birA
                     /note="biotin-protein ligase derived from Escherichia coli;
                     Val2 is a cloning artifact"
                     /protein_id="AEP13897.1"
                     /translation="MVKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLR
                     DWGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKS
                     GDACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLR
                     KLGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQ
                     GWITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLII
                     GDKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     gene            171..1139
                     /gene="birA"
                     /label=birA
     CDS             1167..1211
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     misc_feature    1263..1310
                     /label=T7 terminator
                     /note="T7 terminator"
     terminator      1263..1310
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1481..2272)
                     /codon_start=1
                     /gene="aadA"
                     /product="streptomycin 3-adenylyltransferase"
                     /label=aadA
                     /note="aminoglycoside resistance protein"
                     /protein_id="AEP13899.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            complement(1481..2272)
                     /gene="aadA"
                     /label=aadA
                     /note="derived from correct, original source Tn21"
     promoter        complement(2273..2364)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2412..3150)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="CloDF13 ori"
     rep_origin      complement(2503)
                     /direction=LEFT
     CDS             complement(3360..4442)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /protein_id="AEP13898.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(3360..4442)
                     /gene="lacI"
                     /label=lacI
     promoter        complement(4443..38)
                     /gene="lacI"
                     /label=lacI promoter