LOCUS       sequence-422109-        8959 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Full-length human kinesin-1 with HaloTag at the N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8959)
  AUTHORS   Deguchi T, Iwanski MK, Schentarra EM, Heidebrecht C, Schmidt L, Heck
            J, Weihs T, Schnorrenberg S, Hoess P, Liu S, Chevyreva V, Noh KM, 
            Kapitein LC, Ries J
  TITLE     Direct observation of motor protein stepping in living cells using 
            MINFLUX.
  JOURNAL   Science. 2023 Mar 10;379(6636):1010-1015. doi: 
            10.1126/science.ade2676. Epub 2023 Mar 9.
  PUBMED    36893247
REFERENCE   2  (bases 1 to 8959)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1126/science.ade2676"; journalName: "Science"; date: 
            "2023-03-10- 10"; volume: "379"; issue: "6636"; pages: "1010-1015"
COMMENT     Sequence Label: pHTN_HaloTag_KIF5B
FEATURES             Location/Qualifiers
     source          1..8959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      29..38
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             35..925
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             938..958
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(3878..3896)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3928..4049
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3939..3958)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3993..4012
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4294..4651
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4294..4485
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4317..4337)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4502..4637
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4702..5496
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4756..4775)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5366..5385
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5560..5608
                     /note="synthetic polyadenylation signal"
     promoter        5764..5868
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5869..5937,5938..6729)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6087..6106)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6887..7475)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6967..6986)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7591..7874
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        8065..8444
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8445..8648
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8599..8619
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          8784..8916
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     8899..8918
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 taatacgact cactataggg ctagcaaagc caccatggca gaaatcggta ctggctttcc
       61 attcgacccc cattatgtgg aagtcctggg cgagcgcatg cactacgtcg atgttggtcc
      121 gcgcgatggc acccctgtgc tgttcctgca cggtaacccg acctcctcct acgtgtggcg
      181 caacatcatc ccgcatgttg caccgaccca tcgctgcatt gctccagacc tgatcggtat
      241 gggcaaatcc gacaaaccag acctgggtta tttcttcgac gaccacgtcc gcttcatgga
      301 tgccttcatc gaagccctgg gtctggaaga ggtcgtcctg gtcattcacg actggggctc
      361 cgctctgggt ttccactggg ccaagcgcaa tccagagcgc gtcaaaggta ttgcatttat
      421 ggagttcatc cgccctatcc cgacctggga cgaatggcca gaatttgccc gcgagacctt
      481 ccaggccttc cgcaccaccg acgtcggccg caagctgatc atcgatcaga acgtttttat
      541 cgagggtacg ctgccgatgg gtgtcgtccg cccgctgact gaagtcgaga tggaccatta
      601 ccgcgagccg ttcctgaatc ctgttgaccg cgagccactg tggcgcttcc caaacgagct
      661 gccaatcgcc ggtgagccag cgaacatcgt cgcgctggtc gaagaataca tggactggct
      721 gcaccagtcc cctgtcccga agctgctgtt ctggggcacc ccaggcgttc tgatcccacc
      781 ggccgaagcc gctcgcctgg ccaaaagcct gcctaactgc aaggctgtgg acatcggccc
      841 gggtctgaat ctgctgcaag aagacaaccc ggacctgatc ggcagcgaga tcgcgcgctg
      901 gctgtcgacg ctcgagattt ccggcgagcc aaccactgag gatctgtact ttcagagcga
      961 taacatggcg gacctggccg agtgcaacat caaagtgatg tgtcgcttca gacctctcaa
     1021 cgagtctgaa gtgaaccgcg gcgacaagta catcgccaag tttcagggag aagacacggt
     1081 cgtgatcgcg tccaagcctt atgcatttga tcgggtgttc cagtcaagca catctcaaga
     1141 gcaagtgtat aatgactgtg caaagaagat tgttaaagat gtacttgaag gatataatgg
     1201 aacaatattt gcatatggac aaacatcctc tgggaagaca cacacaatgg agggtaaact
     1261 tcatgatcca gaaggcatgg gaattattcc aagaatagtg caagatattt ttaattatat
     1321 ttactccatg gatgaaaatt tggaatttca tattaaggtt tcatattttg aaatatattt
     1381 ggataagata agggacctgt tagatgtttc aaagaccaac ctttcagttc atgaagacaa
     1441 aaaccgagtt ccctatgtaa aggggtgcac agagcgtttt gtatgtagtc cagatgaagt
     1501 tatggatacc atagatgaag gaaaatccaa cagacatgta gcagttacaa atatgaatga
     1561 acatagctct aggagtcaca gtatatttct tattaatgtc aaacaagaga acacacaaac
     1621 ggaacaaaag ctgagtggaa aactttatct ggttgattta gctggtagtg aaaaggttag
     1681 taaaactgga gctgaaggtg ctgtgctgga tgaagctaaa aacatcaaca agtcactttc
     1741 tgctcttgga aatgttattt ctgctttggc tgagggtagt acatatgttc catatcgaga
     1801 tagtaaaatg acaagaatcc ttcaagattc attaggtggc aactgtagaa ccactattgt
     1861 aatttgctgc tctccatcat catacaatga gtctgaaaca aaatctacac tcttatttgg
     1921 ccaaagggcc aaaacaatta agaacacagt ttgtgtcaat gtggagttaa ctgcagaaca
     1981 gtggaaaaag aagtatgaaa aagaaaaaga aaaaaataag atcctgcgga acactattca
     2041 gtggcttgaa aatgagctca acagatggcg taatggggag acggtgccta ttgatgaaca
     2101 gtttgacaaa gagaaagcca acttggaagc tttcacagtg gataaagata ttactcttac
     2161 caatgataaa ccagcaaccg caattggagt tataggaaat tttactgatg ctgaaagaag
     2221 aaagtgtgaa gaagaaattg ctaaattata caaacagctt gatgacaagg atgaagaaat
     2281 taaccagcaa agtcaactgg tagagaaact gaagacgcaa atgttggatc aggaggagct
     2341 tttggcatct accagaaggg atcaagacaa tatgcaagct gagctgaatc gccttcaagc
     2401 agaaaatgat gcctctaaag aagaagtgaa agaagtttta caggccctag aagaacttgc
     2461 tgtcaattat gatcagaagt ctcaggaagt tgaagacaaa actaaggaat atgaattgct
     2521 tagtgatgaa ttgaatcaga aatcggcaac tttagcgagt atagatgctg agcttcagaa
     2581 acttaaggaa atgaccaacc accagaaaaa acgagcagct gagatgatgg catctttact
     2641 aaaagacctt gcagaaatag gaattgctgt gggaaataat gatgtaaagc agcctgaggg
     2701 aactggcatg atagatgaag agttcactgt tgcaagactc tacattagca aaatgaagtc
     2761 agaagtaaaa accatggtga aacgttgcaa gcagttagaa agcacacaaa ctgagagcaa
     2821 caaaaaaatg gaagaaaatg aaaaggagtt agcagcatgt cagcttcgta tctctcaaca
     2881 tgaagccaaa atcaagtcat tgactgaata ccttcaaaat gtggaacaaa agaaaagaca
     2941 gttggaggaa tctgtcgatg ccctcagtga agaactagtc cagcttcgag cacaagagaa
     3001 agtccatgaa atggaaaagg agcacttaaa taaggttcag actgcaaatg aagttaagca
     3061 agctgttgaa cagcagatcc agagccatag agaaactcat caaaaacaga tcagtagttt
     3121 gagagatgaa gtagaagcaa aagcaaaact tattactgat cttcaagacc aaaaccagaa
     3181 aatgatgtta gagcaggaac gtctaagagt agaacatgag aagttgaaag ccacagatca
     3241 ggaaaagagc agaaaactac atgaacttac ggttatgcaa gatagacgag aacaagcaag
     3301 acaagacttg aagggtttgg aagagacagt ggcaaaagaa cttcagactt tacacaacct
     3361 gcgcaaactc tttgttcagg acctggctac aagagttaaa aagagtgctg agattgattc
     3421 tgatgacacc ggaggcagcg ctgctcagaa gcaaaaaatc tcctttcttg aaaataatct
     3481 tgaacagctc actaaagtgc acaaacagtt ggtacgtgat aatgcagatc tccgctgtga
     3541 acttcctaag ttggaaaagc gacttcgagc tacagctgag agagtgaaag ctttggaatc
     3601 agcactgaaa gaagctaaag aaaatgcatc tcgtgatcgc aaacgctatc agcaagaagt
     3661 agatcgcata aaggaagcag tcaggtcaaa gaatatggcc agaagagggc attctgcaca
     3721 gattgctaaa cctattcgtc ccgggcaaca tccagcagct tctccaactc acccaagtgc
     3781 aattcgtgga ggaggtgcat ttgttcagaa cagccagcca gtggcagtgc gaggtggagg
     3841 aggcaaacaa gtgtaattgc tgagttggct gctgccaccg ctgagcaata actagcataa
     3901 ccccttggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     3961 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     4021 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     4081 aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta
     4141 aaatcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     4201 cgcatctgtg cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat
     4261 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa
     4321 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4381 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4441 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4501 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4561 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4621 aggctttttt ggaggcctag gcttttgcaa aaagcttgat tcttctgaca caacagtctc
     4681 gaacttaagg ctagagccac catgattgaa caagatggat tgcacgcagg ttctccggcc
     4741 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4801 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4861 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4921 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4981 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5041 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5101 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5161 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5221 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5281 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5341 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5401 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5461 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5521 ccgaccaagc gacgcccaac ctgccatcac gatggccgca ataaaatatc tttattttca
     5581 ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga taaggatcct ctttgcgctt
     5641 gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc
     5701 tgcggactgg ctttctacgt aatggtttct tagacgtcag gtggcacttt tcggggaaat
     5761 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5821 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5881 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5941 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     6001 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     6061 ccaatgatga gcactttcaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     6121 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     6181 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     6241 ataaccatga gtgataacac tgcggccaac ttacttctga caactatcgg aggaccgaag
     6301 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6361 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     6421 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6481 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6541 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6601 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6661 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6721 cattggtaat tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg
     6781 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6841 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6901 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6961 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     7021 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7081 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7141 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7201 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7261 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7321 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7381 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7441 agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg
     7501 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7561 aaaggatctt cacctagatc cttttatagt ccggaaatac aggaacgcac gctggatggc
     7621 ccttcgctgg gatggtgaaa ccatgaaaaa tggcagcttc agtggattaa gtgggggtaa
     7681 tgtggcctgt accctctggt tgcataggta ttcatacggt taaaatttat caggcgcgat
     7741 tgcggcagtt tttcgggtgg tttgttgcca tttttacctg tctgctgccg tgatcgcgct
     7801 gaacgcgttt tagcggtgcg tacaattaag ggattatggt aaatccactt actgtctgcc
     7861 ctcgtagcca tcgagataaa ccgcagtact ccggccacga tgcgtccggc gtagaggatc
     7921 gagatcttca atattggcca ttagccatat tattcattgg ttatatagca taaatcaata
     7981 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
     8041 catgtccaat atgaccgcca tgttggcatt gattattgac tagttattaa tagtaatcaa
     8101 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     8161 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     8221 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     8281 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc cctattgacg
     8341 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta cgggactttc
     8401 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     8461 agtacaccaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     8521 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     8581 ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     8641 gcagagctgg tttagtgaac cgtcagatca ctagaagctt tattgcggta gtttatcaca
     8701 gttaaattgc taacgcagtc agtgcttctg acacaacagt ctcgaactta agctgcagaa
     8761 gttggtcgtg aggcactggg caggtaagta tcaaggttac aagacaggtt taaggagacc
     8821 aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg
     8881 gtcttactga catccacttt gcctttctct ccacaggtgt ccactcccag ttcaattaca
     8941 gctcttaagg ctagagtat
//
