LOCUS       40924_34729        6372 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pPK115, complete sequence.
ACCESSION   .
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     Kinesin's 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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6372)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Cell 
            Biol."; date: "1999"; volume: "1"; issue: "5"; pages: "288-292"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-DEC-1998) Department of Physiology and Biophysics, University of
            Washington, Campus Box 357290, Seattle, WA 98195-7290, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        494..598
                     /label=AmpR promoter
     CDS             599..1456
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1630..2218
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2404..2546)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2651..2839)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        3348..3366
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3398..3420
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     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
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     CDS             6176..6193
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             6216..6233
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      6300..6347
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
      601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1141 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2341 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2401 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg
     2461 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     2521 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     2581 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2641 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     2701 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     2761 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     2821 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     2881 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     2941 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     3001 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     3061 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     3121 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     3181 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     3241 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     3301 cccgtggcca ggacccaacg ctgcccgaga tctcgatccc gcgaaattaa tacgactcac
     3361 tatagggaga ccacaacggt ttccctctag aaataatttt gtttaacttt aagaaggaga
     3421 tatacatatg agcgcagaac gagaaattcc agccgaggac agcatcaaag tggtctgccg
     3481 attccgaccg ctgaacgaca gcgaagagaa ggccggctcc aagttcgtgg tcaagttccc
     3541 caacaatgtg gaggagaact gcatatccat agcgggcaag gtgtatttgt tcgacaaggt
     3601 cttcaaaccg aatgcatccc aggaaaaggt ctacaatgag gcggccaagt ccattgttac
     3661 ggatgtcctg gccgggtaca atggaacgat attcgcatac ggtcagacgt cctccggaaa
     3721 aacgcatacg atggagggcg tgatcgggga ctccgtaaaa cagggtatca tacctaggat
     3781 cgtcaacgac attttcaatc acatctacgc gatggaggtg aacctggagt ttcacatcaa
     3841 ggtctcctac tacgagatct acatggacaa gattcgagat ctgttggacg tctccaaggt
     3901 gaaccttagt gtgcacgagg ataagaaccg ggtgccgtac gtcaagggcg ctacggaacg
     3961 gttcgtctcg tcgccggagg atgttttcga ggtgatcgag gagggcaaat ccaatcgtca
     4021 catcgctgtg acaaacatga acgagcattc ttcgcgatcc cactcagtat tccttatcaa
     4081 tgtgaagcag gagaacctgg agaaccagaa gaaactatcc ggcaaactct acctggtgga
     4141 tttggccgga tccgagaagg tttccaagac tggagcggag ggaaccgttc ttgatgaagc
     4201 caagaacatc aacaagtcgc tgtcggcctt gggcaacgta atttctgccc tggcggacgg
     4261 aaacaaaacg cacatcccct accgtgattc caagctaacg cgcatcctgc aggagtcgct
     4321 gggaggcaac gcacgcacaa ccatcgtcat ctgctgctct ccagccagtt tcaacgaatc
     4381 tgaaacgaag tcaacgctgg acttcggtcg tagagccaag acagtgaaga acgtggtctg
     4441 cgttaacgag gagcttactg ccgaggaatg gaagcgacgc tatgaaaagg agaaggaaaa
     4501 gaacgcccga ctaaagggta aggtggagaa gcttgagatc gagcttgcgc gctggagagc
     4561 gggtgaaact gttaaggcgg aggagcaaat caacatggag gatctcatgg aggcaagcac
     4621 gcccaacctg gaagtggagg cagcacagac ggcggcggcc gaggccgctt tggccgccca
     4681 gcgaacggct ctcgccaata tgtccgcatc ggttgccgtg aacgagcagg ccaggctggc
     4741 tacagagtgc gagcgtctct accagcagct ggacgacaag gatgaggaga tcaatcagca
     4801 gagccagtac gccgagcagc tcaaggagca ggtgatggag caggaggaac tcatcgctaa
     4861 cgctcggcgt gagtatgaga ctttgcagtc ggagatggcg cgaattcaac aggagaacga
     4921 gtccgccaag gaagaggtta aggaggtgct ccaagctctc gaagagctga ctgtaaacta
     4981 cgaccagaaa tcccaggaga tcgataacaa gaacaaggat atcgatgccc tcaacgagga
     5041 gctgcagcag aagcagtctg tgttcaacgc cgcctccaca gagctacagc agctcaagga
     5101 catggccaag aacattgccc agcgatgctc caacatggaa acacagcagg ctgactccaa
     5161 caagaagatc tccgaatatg agaaagatct gggcgagtac cggctactca tttcgcagca
     5221 cgaggcacgc atgaagtcgc tgcaggagtc gatgcgggag gcagagaaca agaagcgcac
     5281 gctcgaggaa caaatcgatt cgctgcgcga ggaatgcgcc aagctcaagg ccgccgagca
     5341 cgtttccgcc gttaacgccg aggagaaaca gcgggctgag gagctgcgct ccatgttcga
     5401 ttctcagatg gacgagctac gcgaagccca cacccggcag gtgtccgagc tccgggacga
     5461 aattgccgcc aagcagcacg aaatggacga gatgaaggat gtccatcaaa agctgctctt
     5521 ggcgcaccaa cagatgacgg ccgactacga gaaggtgcgc caggaggatg ccgagaagtc
     5581 cagcgagctt cagaacatca tcctcaccaa cgagcgtcgg gagcaagcgc gcaaagacct
     5641 caagggcctg gaggacacgg tggccaagga gttgcagacg ctacacaacc tgcgaaaact
     5701 tttcgttcag gatctacagc aacgaatccg aaagaatgtc gtaaacgagg agagcgagga
     5761 ggacggtgga tcactcgcgc agaaacagaa gatttccttc ttggagaaca accttgatca
     5821 gctgaccaag gtgcacaagc aattggtgcg ggacaacgcc gatctgcggt gcgagctgcc
     5881 caagctggag aagcgtctac gctgtaccat ggagcgggtg aaagctctgg agacagcgct
     5941 caaggaggcg aaggagggcg caatgcggga tcgcaagcgc taccaatacg aggtggaccg
     6001 catcaaggag gcggtgcgac agaagcatct gggcagacgt ggcccacagg cacagatcgc
     6061 aaagccgatc cggtccggcc aaggtgcaat cgccattcgt ggtggtggcg ccgttggagg
     6121 accatccccg ctggcccagg ttaatcctgt caactcgctt gttccgcgtg ggagccatca
     6181 tcatcatcat cactaagcgg ccgcactcga tcgagcacca ccaccaccac cactgagatc
     6241 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
     6301 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
     6361 ctatatccgg at
//
