LOCUS       Exported                6372 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pPK115, complete sequence.
ACCESSION   AF117643
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6372)
  AUTHORS   Coy DL, Hancock WO, Wagenbach M, Howard J.
  TITLE     Kinesins tail domain is an inhibitory regulator of the motor domain
  JOURNAL   Nat. Cell Biol. 1 (5), 288-292 (1999)
  PUBMED    10559941
REFERENCE   2  (bases 1 to 6372)
  AUTHORS   Coy DL, Wagenbach M, Howard J.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-DEC-1998) Department of Physiology and Biophysics, 
            University of Washington, Campus Box 357290, Seattle, WA 98195-7290,
            USA
REFERENCE   3  (bases 1 to 6372)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6372
                     /organism="Expression vector pPK115"
                     /mol_type="other DNA"
                     /note="derived from Expression vector pPK113 presented in 
                     GenBank Accession Number AF053733"
                     /db_xref="taxon:91184"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        494..598
                     /gene="bla"
                     /label=AmpR promoter
     CDS             599..1459
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance marker"
                     /protein_id="AAD29959.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            599..1459
                     /gene="bla"
                     /label=bla
     rep_origin      1630..2218
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2404..2546
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2648..2839)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        3348..3366
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3398..3420
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3428..6196
                     /codon_start=1
                     /product="kinesin delta 560-624"
                     /label=kinesin delta 560-624
                     /note="chimera of mutated Drosophila melanogaster kinesin 
                     heavy chain C-terminal region and histidine tag"
                     /protein_id="AAD29958.1"
                     /translation="MSAEREIPAEDSIKVVCRFRPLNDSEEKAGSKFVVKFPNNVEENC
                     ISIAGKVYLFDKVFKPNASQEKVYNEAAKSIVTDVLAGYNGTIFAYGQTSSGKTHTMEG
                     VIGDSVKQGIIPRIVNDIFNHIYAMEVNLEFHIKVSYYEIYMDKIRDLLDVSKVNLSVH
                     EDKNRVPYVKGATERFVSSPEDVFEVIEEGKSNRHIAVTNMNEHSSRSHSVFLINVKQE
                     NLENQKKLSGKLYLVDLAGSEKVSKTGAEGTVLDEAKNINKSLSALGNVISALADGNKT
                     HIPYRDSKLTRILQESLGGNARTTIVICCSPASFNESETKSTLDFGRRAKTVKNVVCVN
                     EELTAEEWKRRYEKEKEKNARLKGKVEKLEIELARWRAGETVKAEEQINMEDLMEASTP
                     NLEVEAAQTAAAEAALAAQRTALANMSASVAVNEQARLATECERLYQQLDDKDEEINQQ
                     SQYAEQLKEQVMEQEELIANARREYETLQSEMARIQQENESAKEEVKEVLQALEELTVN
                     YDQKSQEIDNKNKDIDALNEELQQKQSVFNAASTELQQLKDMAKNIAQRCSNMETQQAD
                     SNKKISEYEKDLGEYRLLISQHEARMKSLQESMREAENKKRTLEEQIDSLREECAKLKA
                     AEHVSAVNAEEKQRAEELRSMFDSQMDELREAHTRQVSELRDEIAAKQHEMDEMKDVHQ
                     KLLLAHQQMTADYEKVRQEDAEKSSELQNIILTNERREQARKDLKGLEDTVAKELQTLH
                     NLRKLFVQDLQQRIRKNVVNEESEEDGGSLAQKQKISFLENNLDQLTKVHKQLVRDNAD
                     LRCELPKLEKRLRCTMERVKALETALKEAKEGAMRDRKRYQYEVDRIKEAVRQKHLGRR
                     GPQAQIAKPIRSGQGAIAIRGGGAVGGPSPLAQVNPVNSLVPRGSHHHHHH"
     RBS             4939..4947
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     CDS             6158..6175
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             6176..6193
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6216..6233
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      6300..6347
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"