LOCUS       sequence-100639-        5406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Non-fluorescent, truncated kinesin-1 fused to EF Hand linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5406)
  AUTHORS   Norris SR, Soppina V, Dizaji AS, Schimert KI, Sept D, Cai D, 
            Sivaramakrishnan S, Verhey KJ
  TITLE     A method for multiprotein assembly in cells reveals independent 
            action of kinesins in complex.
  JOURNAL   J Cell Biol. 2014 Nov 10;207(3):393-406. doi: 10.1083/jcb.201407086.
            Epub 2014 Nov 3.
  PUBMED    25365993
REFERENCE   2  (bases 1 to 5406)
  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: 
            "10.1083/jcb.201407086"; journalName: "J Cell Biol"; date: 
            "2014-11-10- 10"; volume: "207"; issue: "3"; pages: "393-406"
COMMENT     Sequence Label: KIF5C(1-560)-EF(C)
FEATURES             Location/Qualifiers
     source          1..5406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        34..133
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
     primer_bind     84..104
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1078..1089
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    2167..2288
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2204..2223)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2258..2277
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2295..2750)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2432..2453)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2644..2663
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2777..2881
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2883..3240
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2883..3074
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2906..2926)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3091..3226
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3275..4069
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3275..4069
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3329..3348)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3939..3958
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4257..4276)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4301..4348
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4677..5265
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5166..5185
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
       61 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      121 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatctc
      181 gagctcaagc ttcgaattca tggcggatcc agccgaatgc agcatcaaag tgatgtgccg
      241 gttccggccc ctcaacgaag ctgagatcct ccgcggggac aaattcatcc ccaaattcaa
      301 aggcgaggag acggtggtga tcgggcaagg gaagccgtat gtctttgacc gagttctgcc
      361 gcccaacaca acccaggagc aggtctacaa tgcctgtgcg aagcagattg tcaaagatgt
      421 cctcgagggt tacaatggga caatttttgc atacgggcag acttcatcag gaaaaactca
      481 taccatggag gggaagctgc acgatcctca gcttatgggt atcataccca ggatcgcaca
      541 cgatattttc gatcacatct attccatgga tgagaacttg gagtttcata tcaaggtttc
      601 ctattttgag atctacttgg acaaaataag ggacttgctt gacgtgtcca agaccaactt
      661 ggctgttcat gaagacaaaa acagagtccc ctatgtaaag gggtgcaccg agaggtttgt
      721 gtcaagcccg gaggaggtca tggatgtgat cgatgagggc aaagcaaacc gacatgtggc
      781 tgtgacaaac atgaacgaac acagctccag gagtcacagt atcttcctga ttaacattaa
      841 acaagagaat gtggagactg aaaaaaaact cagcgggaag ctgtatttgg ttgatttggc
      901 tgggagtgaa aaggtcagca aaactggtgc cgagggagct gttcttgatg aagctaaaaa
      961 tatcaacaag tctttgtctg ctcttggaaa tgtgatctct gccttggcag aagggacaaa
     1021 aacacatgtg ccataccggg acagcaagat gactcggatt ctccaggact ctctgggtgg
     1081 gaactgcagg accaccattg tcatttgctg ttctccttca gtcttcaatg aggctgagac
     1141 caagtccacg ctgatgtttg gacagagagc aaagaccatc aagaatacag tctctgtgaa
     1201 cttggaacta acagcagaag aatggaagaa gaaatatgaa aaagagaaag agaagaataa
     1261 ggccctgaag agtgtcatcc agcatctgga ggtggagctg aacaggtgga ggaacgggga
     1321 agctgtgcct gaggatgaac agatcagtgc caaggaccag aagaacctgg agccctgcga
     1381 caacacaccc atcatagaca acatcacacc tgtggttgac ggcatctctg ctgagaaaga
     1441 gaagtatgac gaggaaatca ccagtctgta ccgacagctg gatgacaagg acgatgaaat
     1501 taaccagcag agccagctgg ctgaaaagct gaagcaacag atgttggatc aggacgagct
     1561 tctagcttcc accagaaggg actacgagaa aatccaggag gagctgaccc gcctccagat
     1621 tgaaaacgag gcggctaaag acgaggtgaa ggaggtcctc caggccctgg aggagctggc
     1681 tgtcaattac gaccagaagt cacaggaagt ggaggacaag acccgggcca acgagcaact
     1741 gaccgatgag ctggcccaga aaacgacgac actgacaacc acccagcggg agctgagtca
     1801 gctacaggag cttagcaacc accagaagaa gagggccacg gagatcctga acctgcttct
     1861 caaggatctg ggcgagctgg taccgggagg tggtggagga ggaggtggag gaccggtttc
     1921 gggatcggga gggagcggag gatccggggg ttccggttcg acgctcgacg aacttttcga
     1981 ggaactggac aagaacggcg atggggaagt cagcttcgag gagtttcaag tgttggtaaa
     2041 gaaaatctcc cagtgatcta gatcataatc agccatacca catttgtaga ggttttactt
     2101 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     2161 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2221 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2281 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     2341 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     2401 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     2461 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     2521 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     2581 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     2641 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     2701 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     2761 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2821 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2881 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     2941 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3001 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3061 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3121 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3181 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3241 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3301 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3361 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     3421 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     3481 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     3541 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     3601 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     3661 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     3721 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     3781 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     3841 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     3901 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     3961 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4021 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4081 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4141 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4201 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     4261 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4321 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     4381 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     4441 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     4501 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     4561 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4621 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4681 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4741 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4801 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     4861 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4921 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4981 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5041 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5101 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5161 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5221 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5281 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5341 tcccctgatt ctgtggataa ccgtattacc gccatgcatc ratdttsday brtarymcmm
     5401 nbckmt
//
