LOCUS       V004079                 9915 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004079
VERSION     V004079
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9915)
  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 9915)
  AUTHORS   Coy DL, Wagenbach M, Howard J.
  TITLE     Mechanochemical Coupling Stoichiometry of Single Kinesin Motors
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 9915)
  AUTHORS   Coy DL, Wagenbach M, Howard J.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-MAR-1998) Physiology and Biophysics, University of
            Washington, Campus Box 357290, Seattle, WA 98195-7290, USA
REFERENCE   4  (bases 1 to 9915)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 9915)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Biol.
            Chem."; date: "1999"; volume: "274"; issue: "6"; pages: "3667-3671"
            SGRef: number: 2; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
            (23-MAR-1998) Physiology and Biophysics, University of Washington,
            Campus Box 357290, Seattle, WA 98195-7290, USA"
            SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9915
                     /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
                     /label="AmpR"
                     /note="beta-lactamase"
     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)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(3614..3635)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3651..4730)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(4731..4808)
                     /label="lacI promoter"
     promoter        5117..5135
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5136..5160
                     /label="lac operator"
                     /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             5175..5197
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5205..8129
                     /gene="Khc"
                     /label="Kinesin heavy chain"
                     /note="Kinesin heavy chain from Drosophila melanogaster.
                     Accession#: P17210"
     CDS             8154..8177
                     /codon_start=1
                     /product="co-translated peptide"
                     /label="co-translated peptide"
                     /note="modeled on Schoner, B.E. et al. (1984) PNAS USA
                     81:5403-5407"
                     /protein_id="AAD13355.1"
                     /translation="MFPLEDD"
     CDS             8178..9701
                     /gene="Klc"
                     /label="Kinesin light chain"
                     /note="Kinesin light chain from Drosophila melanogaster.
                     Accession#: P46824"
     CDS             9702..9719
                     /label="thrombin site"
                     /note="thrombin recognition and cleavage site"
     CDS             9720..9746
                     /label="9xHis"
                     /note="9xHis affinity tag"
     CDS             9759..9776
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      9843..9890
                     /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 cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     3361 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     3421 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     3481 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     3541 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     3601 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     3661 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3721 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     3781 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     3841 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     3901 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
     3961 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     4021 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     4081 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     4141 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     4201 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     4261 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     4321 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     4381 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     4441 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     4501 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     4561 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     4621 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     4681 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     4741 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     4801 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     4861 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     4921 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     4981 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     5041 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
     5101 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
     5161 cctctagaaa taattttgtt taactttaag aaggagatat acatatgagc gcagaacgag
     5221 aaattccagc cgaggacagc atcaaagtgg tctgccgatt ccgaccgctg aacgacagcg
     5281 aagagaaggc cggctccaag ttcgtggtca agttccccaa caatgtggag gagaactgca
     5341 tatccatagc gggcaaggtg tatttgttcg acaaggtctt caaaccgaat gcatcccagg
     5401 aaaaggtcta caatgaggcg gccaagtcca ttgttacgga tgtcctggcc gggtacaatg
     5461 gaacgatatt cgcatacggt cagacgtcct ccggaaaaac gcatacgatg gagggcgtga
     5521 tcggggactc cgtaaaacag ggtatcatac ctaggatcgt caacgacatt ttcaatcaca
     5581 tctacgcgat ggaggtgaac ctggagtttc acatcaaggt ctcctactac gagatctaca
     5641 tggacaagat tcgagatctg ttggacgtct ccaaggtgaa ccttagtgtg cacgaggata
     5701 agaaccgggt gccgtacgtc aagggcgcta cggaacggtt cgtctcgtcg ccggaggatg
     5761 ttttcgaggt gatcgaggag ggcaaatcca atcgtcacat cgctgtgaca aacatgaacg
     5821 agcattcttc gcgatcccac tcagtattcc ttatcaatgt gaagcaggag aacctggaga
     5881 accagaagaa actatccggc aaactctacc tggtggattt ggccggatcc gagaaggttt
     5941 ccaagactgg agcggaggga accgttcttg atgaagccaa gaacatcaac aagtcgctgt
     6001 cggccttggg caacgtaatt tctgccctgg cggacggaaa caaaacgcac atcccctacc
     6061 gtgattccaa gctaacgcgc atcctgcagg agtcgctggg aggcaacgca cgcacaacca
     6121 tcgtcatctg ctgctctcca gccagtttca acgaatctga aacgaagtca acgctggact
     6181 tcggtcgtag agccaagaca gtgaagaacg tggtctgcgt taacgaggag cttactgccg
     6241 aggaatggaa gcgacgctat gaaaaggaga aggaaaagaa cgcccgacta aagggtaagg
     6301 tggagaagct tgagatcgag cttgcgcgct ggagagcggg tgaaactgtt aaggcggagg
     6361 agcaaatcaa catggaggat ctcatggagg caagcacgcc caacctggaa gtggaggcag
     6421 cacagacggc ggcggccgag gccgctttgg ccgcccagcg aacggctctc gccaatatgt
     6481 ccgcatcggt tgccgtgaac gagcaggcca ggctggctac agagtgcgag cgtctctacc
     6541 agcagctgga cgacaaggat gaggagatca atcagcagag ccagtacgcc gagcagctca
     6601 aggagcaggt gatggagcag gaggaactca tcgctaacgc tcggcgtgag tatgagactt
     6661 tgcagtcgga gatggcgcga attcaacagg agaacgagtc cgccaaggaa gaggttaagg
     6721 aggtgctcca agctctcgaa gagctgactg taaactacga ccagaaatcc caggagatcg
     6781 ataacaagaa caaggatatc gatgccctca acgaggagct gcagcagaag cagtctgtgt
     6841 tcaacgccgc ctccacagag ctacagcagc tcaaggacat gtcctcacac cagaagaagc
     6901 gcatcacgga aatgctaacc aacctactgc gcgacctcgg cgaagtgggc caggccattg
     6961 cccccggcga gtccagcatc gaccttaaga tgagtgctct ggctggcacg gatgccagca
     7021 aggtggagga agatttcacc atggcgcgtt tgtttatcag caagatgaag acggaggcca
     7081 agaacattgc ccagcgatgc tccaacatgg aaacacagca ggctgactcc aacaagaaga
     7141 tctccgaata tgagaaagat ctgggcgagt accggctact catttcgcag cacgaggcac
     7201 gcatgaagtc gctgcaggag tcgatgcggg aggcagagaa caagaagcgc acgctcgagg
     7261 aacaaatcga ttcgctgcgc gaggaatgcg ccaagctcaa ggccgccgag cacgtttccg
     7321 ccgttaacgc cgaggagaaa cagcgggctg aggagctgcg ctccatgttc gattctcaga
     7381 tggacgagct acgcgaagcc cacacccggc aggtgtccga gctccgggac gaaattgccg
     7441 ccaagcagca cgaaatggac gagatgaagg atgtccatca aaagctgctc ttggcgcacc
     7501 aacagatgac ggccgactac gagaaggtgc gccaggagga tgccgagaag tccagcgagc
     7561 ttcagaacat catcctcacc aacgagcgtc gggagcaagc gcgcaaagac ctcaagggcc
     7621 tggaggacac ggtggccaag gagttgcaga cgctacacaa cctgcgaaaa cttttcgttc
     7681 aggatctaca gcaacgaatc cgaaagaatg tcgtaaacga ggagagcgag gaggacggtg
     7741 gatcactcgc gcagaaacag aagatttcct tcttggagaa caaccttgat cagctgacca
     7801 aggtgcacaa gcaattggtg cgggacaacg ccgatctgcg gtgcgagctg cccaagctgg
     7861 agaagcgtct acgctgtacc atggagcggg tgaaagctct ggagacagcg ctcaaggagg
     7921 cgaaggaggg cgcaatgcgg gatcgcaagc gctaccaata cgaggtggac cgcatcaagg
     7981 aggcggtgcg acagaagcat ctgggcagac gtggcccaca ggcacagatc gcaaagccga
     8041 tccggtccgg ccaaggtgca atcgccattc gtggtggtgg cgccgttgga ggaccatccc
     8101 cgctggccca ggttaatcct gtcaactcgt aagcggccgc gagggtatta catatgtttc
     8161 cattggagga tgattaaatg actcaaatgt ctcaggacga aattattacc aatactaaaa
     8221 ctgttctgca aggtctggaa gcactgcgtg ttgaacatgt ttctattatg aacggtattg
     8281 cagaagttca gaaggacaat gaaaagtctg acatgctgcg taagaacatc gaaaatattg
     8341 aactgggtct gtctgaagca caagttatga tggcactgac ttcccatctg cagaacatcg
     8401 aagcagaaaa gcacaagctt aagacacagg tgcgccgcct ccaccaggag aacgcctggc
     8461 tccgcgacga gctggccaac actcagcaaa agttccaggc atccgagcag ctggtggccc
     8521 aactagagga ggagaagaag cacctggagt tcatggcgtc cgtgaagaag tacgacgaga
     8581 atcaggagca ggacgacgct tgcgacaagt cccgcaccga tcccgtggtc gagctgttcc
     8641 cggacgagga aaacgaggac agacacaaca tgtcgcccac tccgcccagc cagttcgcca
     8701 accagacttc cggctacgag attccagcgc gtctccgtac tctgcacaac ctggtcattc
     8761 agtatgcatc gcagggaaga tacgaggtag ccgttcccct ctgcaagcaa gcattggaag
     8821 atctcgaaag gacaagtggt cacgaccatc ccgatgtagc aaccatgctg aacattttgg
     8881 ctctcgtgta ccgcgatcag aacaagtata aggaggccgc caatttactc aacgacgctc
     8941 tgtcgattcg aggaaagaca ctgggcgaga accatccagc cgtggcggcc acgttgaaca
     9001 acttggccgt cctctatggc aaacgtggca agtacaaaga tgccgagcca ctgtgcaagc
     9061 gtgccttgga gatccgagag aaggtcctgg gaaaggatca tcccgatgtt gccaagcaac
     9121 ttaacaatct cgccctgctc tgccagaatc agggcaaata cgacgaggtt gagaagtact
     9181 accagcgagc tctcgacatc tacgaatcaa agttgggtcc cgatgatccc aacgtggcca
     9241 agacaaagaa taatctggcc ggctgctatc tgaagcaagg tagatacacc gaagctgaaa
     9301 tcctctataa gcaggtcttg acgcgagccc acgaacgtga gtttggagcc atcgacagca
     9361 aaaacaagcc catttggcag gtggctgagg agcgtgagga gcacaaattc gataacaggg
     9421 agaacactcc atatggcgag tacggcggtt ggcataaggc cgctaaagtg gattcgccca
     9481 cggtcacaac cactctaaaa aatctgggag cactttaccg acgtcaaggc atgtttgaag
     9541 cggccgaaac cctggaagac tgtgcaatgc ggagtaaaaa agaagcctac gatctagcta
     9601 aacaaacgaa gctctcacaa ttgctaacgt caaacgaaaa gcgacgatcg aaggcaatta
     9661 aagaggattt ggatttctct gaggagaaga atgcgaaacc acttgtgcca cgcgggagcc
     9721 atcaccatca ccatcatcac catcaccatt aagtcgagca ccaccaccac caccactgag
     9781 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     9841 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     9901 gaactatatc cggat
//
