LOCUS       Exported                6567 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pPK113, complete sequence.
ACCESSION   AF053733
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6567)
  AUTHORS   Coy DL, Wagenbach M, Howard J.
  TITLE     Kinesin takes one 8-nm step for each ATP that it hydrolyzes
  JOURNAL   J. Biol. Chem. 274 (6), 3667-3671 (1999)
  PUBMED    9920916
REFERENCE   2  (bases 1 to 6567)
  AUTHORS   Coy DL, Wagenbach M, Howard J.
  TITLE     Mechanochemical Coupling Stoichiometry of Single Kinesin Motors
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6567)
  AUTHORS   Coy DL, Wagenbach M, Howard J.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-MAR-1998) Physiology and Biophysics, University of 
            Washington, Campus Box 357290, Seattle, WA 98195-7290, USA
REFERENCE   4  (bases 1 to 6567)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6567
                     /organism="Expression vector pPK113"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:74548"
     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
     gene            559..1459
                     /gene="bla"
                     /label=bla
     CDS             599..1459
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance marker"
                     /protein_id="AAD13350.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     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"
     regulatory      3348..3364
                     /regulatory_class="promoter"
                     /standard_name="T7 promoter"
     RBS             3398..3420
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3428..6391
                     /codon_start=1
                     /gene="dhk"
                     /product="recombinant kinesin heavy chain"
                     /label=dhk
                     /note="Drosophila melanogaster kinesin heavy chain"
                     /protein_id="AAD13351.1"
                     /translation="MSAEREIPAEDSIKVVCRFRPLNDSEEKAGSKFVVKFPNNVEENC
                     ISIAGKVYLFDKVFKPNASQEKVYNEAAKSIVTDVLAGYNGTIFAYGQTSSGKTHTMEG
                     VIGDSVKQGIIPRIVNDIFNHIYAMEVNLEFHIKVSYYEIYMDKIRDLLDVSKVNLSVH
                     EDKNRVPYVKGATERFVSSPEDVFEVIEEGKSNRHIAVTNMNEHSSRSHSVFLINVKQE
                     NLENQKKLSGKLYLVDLAGSEKVSKTGAEGTVLDEAKNINKSLSALGNVISALADGNKT
                     HIPYRDSKLTRILQESLGGNARTTIVICCSPASFNESETKSTLDFGRRAKTVKNVVCVN
                     EELTAEEWKRRYEKEKEKNARLKGKVEKLEIELARWRAGETVKAEEQINMEDLMEASTP
                     NLEVEAAQTAAAEAALAAQRTALANMSASVAVNEQARLATECERLYQQLDDKDEEINQQ
                     SQYAEQLKEQVMEQEELIANARREYETLQSEMARIQQENESAKEEVKEVLQALEELTVN
                     YDQKSQEIDNKNKDIDALNEELQQKQSVFNAASTELQQLKDMSSHQKKRITEMLTNLLR
                     DLGEVGQAIAPGESSIDLKMSALAGTDASKVEEDFTMARLFISKMKTEAKNIAQRCSNM
                     ETQQADSNKKISEYEKDLGEYRLLISQHEARMKSLQESMREAENKKRTLEEQIDSLREE
                     CAKLKAAEHVSAVNAEEKQRAEELRSMFDSQMDELREAHTRQVSELRDEIAAKQHEMDE
                     MKDVHQKLLLAHQQMTADYEKVRQEDAEKSSELQNIILTNERREQARKDLKGLEDTVAK
                     ELQTLHNLRKLFVQDLQQRIRKNVVNEESEEDGGSLAQKQKISFLENNLDQLTKVHKQL
                     VRDNADLRCELPKLEKRLRCTMERVKALETALKEAKEGAMRDRKRYQYEVDRIKEAVRQ
                     KHLGRRGPQAQIAKPIRSGQGAIAIRGGGAVGGPSPLAQVNPVNSLVPRGSHHHHHH"
     gene            3428..6391
                     /gene="dhk"
                     /label=dhk
     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             6353..6370
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             6371..6388
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6411..6428
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      6495..6542
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"