LOCUS       Exported                3474 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pPANK, complete sequence.
ACCESSION   AY327140
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3474)
  AUTHORS   Binz HK, Stumpp MT, Forrer P, Amstutz P, Pluckthun A.
  TITLE     Designing repeat proteins: well-expressed, soluble and stable 
            proteins from combinatorial libraries of consensus ankyrin repeat 
            proteins
  JOURNAL   J. Mol. Biol. 332 (2), 489-503 (2003)
  PUBMED    12948497
REFERENCE   2  (bases 1 to 3474)
  AUTHORS   Binz KH, Stumpp MT, Forrer P, Amstutz P, Zahnd C, Pluckthun A.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JUN-2003) Biochemistry, University of Zurich, 
            Winterthurerstrasse 190, Zurich 8057, Switzerland
REFERENCE   3  (bases 1 to 3474)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3474
                     /organism="Expression vector pPANK"
                     /mol_type="other DNA"
                     /note="similar to pQE30 (QIAgen) but with mutated BpiI and 
                     BsaI sites"
                     /db_xref="taxon:244215"
     regulatory      7..87
                     /regulatory_class="promoter"
                     /note="T5 promoter"
     promoter        10..54
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /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)."
     protein_bind    62..78
                     /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)."
     RBS             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    115..144
                     /label=RGS-His-tag
                     /note="RGS-His-tag"
     CDS             127..144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    145..198
                     /note="mcs; multiple cloning site"
     terminator      208..302
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             346..1005
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     terminator      1070..1156
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    1324..1464
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      1597..2253
                     /label=ColE1
                     /note="ColE1"
     rep_origin      complement(1650..2238)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2409..3269)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampicillin resistance"
                     /protein_id="AAQ68222.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQAAMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3270..3374)
                     /gene="bla"
                     /label=AmpR promoter