LOCUS       sequence-422108-        7750 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Truncated 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 7750)
  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 7750)
  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[1-560]
FEATURES             Location/Qualifiers
     source          1..7750
                     /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(2669..2687)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2719..2840
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2730..2749)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2784..2803
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3085..3442
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3085..3276
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3108..3128)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3293..3428
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3493..4287
                     /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(3547..3566)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4157..4176
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4351..4399
                     /note="synthetic polyadenylation signal"
     promoter        4555..4659
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4660..4728,4729..5520)
                     /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(4878..4897)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5678..6266)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5758..5777)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6382..6665
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        6856..7235
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7236..7439
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7390..7410
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          7575..7707
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     7690..7709
                     /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 aaaataattg ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggc
     2701 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     2761 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     2821 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     2881 gggagatgtg ggaggttttt ttaagcaagt aaaacctcta caaatgtggt aaaatcgaat
     2941 tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     3001 gcggtatttc acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga
     3061 aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca
     3121 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3181 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3241 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3301 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3361 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3421 tggaggccta ggcttttgca aaaagcttga ttcttctgac acaacagtct cgaacttaag
     3481 gctagagcca ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3541 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3601 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3661 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3721 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3781 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3841 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3901 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3961 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4021 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4081 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4141 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4201 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4261 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4321 cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg
     4381 tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc tctttgcgct tgcgttttcc
     4441 cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg
     4501 gctttctacg taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4561 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4621 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4681 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4741 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4801 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4861 agcactttca aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4921 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4981 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5041 agtgataaca ctgcggccaa cttacttctg acaactatcg gaggaccgaa ggagctaacc
     5101 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5161 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5221 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5281 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5341 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5401 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5461 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5521 ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac
     5581 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5641 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5701 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5761 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5821 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5881 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5941 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6001 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6061 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     6121 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6181 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6241 ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg
     6301 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6361 tcacctagat ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg
     6421 ggatggtgaa accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg
     6481 taccctctgg ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt
     6541 ttttcgggtg gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt
     6601 ttagcggtgc gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc
     6661 atcgagataa accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatcttc
     6721 aatattggcc attagccata ttattcattg gttatatagc ataaatcaat attggctatt
     6781 ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa
     6841 tatgaccgcc atgttggcat tgattattga ctagttatta atagtaatca attacggggt
     6901 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     6961 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     7021 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     7081 acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg
     7141 gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc
     7201 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca
     7261 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     7321 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg
     7381 ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     7441 gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac agttaaattg
     7501 ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcaga agttggtcgt
     7561 gaggcactgg gcaggtaagt atcaaggtta caagacaggt ttaaggagac caatagaaac
     7621 tgggcttgtc gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg
     7681 acatccactt tgcctttctc tccacaggtg tccactccca gttcaattac agctcttaag
     7741 gctagagtat
//
