LOCUS       sequence-368397-        6608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression of truncated version (aa 1-560) of rat 
            kinesin-1 KIF5C tagged with Halo and Flag tags.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6608)
  AUTHORS   Budaitis BG, Badieyan S, Yue Y, Blasius TL, Reinemann DN, Lang MJ, 
            Cianfrocco MA, Verhey KJ
  TITLE     A kinesin-1 variant reveals motor-induced microtubule damage in 
            cells.
  JOURNAL   Curr Biol. 2022 Jun 6;32(11):2416-2429.e6. doi: 
            10.1016/j.cub.2022.04.020. Epub 2022 May 2.
  PUBMED    35504282
REFERENCE   2  (bases 1 to 6608)
  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: 
            "10.1016/j.cub.2022.04.020"; journalName: "Curr Biol"; date: 
            "2022-06-6- 6"; volume: "32"; issue: "11"; pages: "2416-2429.e6"
COMMENT     Sequence Label: pN1-RnKIF5C(1-560)-Halo-Flag
FEATURES             Location/Qualifiers
     source          1..6608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1513..1524
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     regulatory      2342..2351
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3248..3271
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3396..3517
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3433..3452)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3487..3506
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3524..3979)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3661..3682)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3873..3892
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4006..4110
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4112..4469
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4112..4303
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4135..4155)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4320..4455
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4504..5298
                     /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             4504..5298
                     /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(4558..4577)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5168..5187
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5486..5505)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5530..5577
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5906..6494
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6395..6414
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcatggcg gatccagccg aatgcagcat
      661 caaagtgatg tgccggttcc ggcccctcaa cgaagctgag atcctccgcg gggacaaatt
      721 catccccaaa ttcaaaggcg aggagacggt ggtgatcggg caagggaagc cgtatgtctt
      781 tgaccgagtt ctgccgccca acacaaccca ggagcaggtc tacaatgcct gtgcgaagca
      841 gattgtcaaa gatgtcctcg agggttacaa tgggacaatt tttgcatacg ggcagacttc
      901 atcaggaaaa actcatacca tggaggggaa gctgcacgat cctcagctta tgggtatcat
      961 acccaggatc gcacacgata ttttcgatca catctattcc atggatgaga acttggagtt
     1021 tcatatcaag gtttcctatt ttgagatcta cttggacaaa ataagggact tgcttgacgt
     1081 gtccaagacc aacttggctg ttcatgaaga caaaaacaga gtcccctatg taaaggggtg
     1141 caccgagagg tttgtgtcaa gcccggagga ggtcatggat gtgatcgatg agggcaaagc
     1201 aaaccgacat gtggctgtga caaacatgaa cgaacacagc tccaggagtc acagtatctt
     1261 cctgattaac attaaacaag agaatgtgga gactgaaaaa aaactcagcg ggaagctgta
     1321 tttggttgat ttggctggga gtgaaaaggt cagcaaaact ggtgccgagg gagctgttct
     1381 tgatgaagct aaaaatatca acaagtcttt gtctgctctt ggaaatgtga tctctgcctt
     1441 ggcagaaggg acaaaaacac atgtgccata ccgggacagc aagatgactc ggattctcca
     1501 ggactctctg ggtgggaact gcaggaccac cattgtcatt tgctgttctc cttcagtctt
     1561 caatgaggct gagaccaagt ccacgctgat gtttggacag agagcaaaga ccatcaagaa
     1621 tacagtctct gtgaacttgg aactaacagc agaagaatgg aagaagaaat atgaaaaaga
     1681 gaaagagaag aataaggccc tgaagagtgt catccagcat ctggaggtgg agctgaacag
     1741 gtggaggaac ggggaagctg tgcctgagga tgaacagatc agtgccaagg accagaagaa
     1801 cctggagccc tgcgacaaca cacccatcat agacaacatc acacctgtgg ttgacggcat
     1861 ctctgctgag aaagagaagt atgacgagga aatcaccagt ctgtaccgac agctggatga
     1921 caaggacgat gaaattaacc agcagagcca gctggctgaa aagctgaagc aacagatgtt
     1981 ggatcaggac gagcttctag cttccaccag aagggactac gagaaaatcc aggaggagct
     2041 gacccgcctc cagattgaaa acgaggcggc taaagacgag gtgaaggagg tcctccaggc
     2101 cctggaggag ctggctgtca attacgacca gaagtcacag gaagtggagg acaagacccg
     2161 ggccaacgag caactgaccg atgagctggc ccagaaaacg acgacactga caaccaccca
     2221 gcgggagctg agtcagctac aggagcttag caaccaccag aagaagaggg ccacggagat
     2281 cctgaacctg cttctcaagg atctgggcga gctggtaccg cgggcccggg atccaccggt
     2341 cgccaccatg ggttccgaaa tcggtacagg cttccccttc gacccccatt atgtggaagt
     2401 cctgggcgag cgtatgcact acgtcgatgt tggaccgcgg gatggcacgc ctgtgctgtt
     2461 cctgcacggt aacccgacct cgtcctacct gtggcgcaac atcatcccgc atgtagcacc
     2521 gagtcatcgg tgcattgctc cagacctgat cgggatggga aaatcggaca aaccagacct
     2581 cgattatttc ttcgacgacc acgtccgcta cctcgatgcc ttcatcgaag ccttgggttt
     2641 ggaagaggtc gtcctggtca tccacgactg gggctcagct ctcggattcc actgggccaa
     2701 gcgcaatccg gaacgggtca aaggtattgc atgtatggaa ttcatccggc ctatcccgac
     2761 gtgggacgaa tggccagaat tcgcccgtga gaccttccag gccttccgga ccgccgacgt
     2821 cggccgagag ttgatcatcg atcagaacgc tttcatcgag ggtgcgctcc cgatgggggt
     2881 cgtccgtccg cttacggagg tcgagatgga ccactatcgc gagcccttcc tcaagcctgt
     2941 tgaccgagag ccactgtggc gattccccaa cgagctgccc atcgccggtg agcccgcgaa
     3001 catcgtcgcg ctcgtcgagg catacatgaa ctggctgcac cagtcacctg tcccgaagtt
     3061 gttgttctgg ggcacacccg gcgtactgat ccccccggcc gaagccgcga gacttgccga
     3121 aagcctcccc aactgcaaga cagtggacat cggcccggga ttgttcttgc tccaggaaga
     3181 caacccggac cttatcggca gtgagatcgc gcgctggctc ccaggactgg ccggcggtgt
     3241 acaaggagat tacaaggatg acgatgacaa gtgagcggcc gcgactctag atcataatca
     3301 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     3361 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     3421 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3481 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     3541 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     3601 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     3661 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     3721 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     3781 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     3841 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     3901 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     3961 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4021 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4081 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     4141 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4201 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4261 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4321 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4381 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4441 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     4501 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4561 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4621 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4681 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4741 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4801 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4861 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4921 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4981 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     5041 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5101 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5161 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5221 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5281 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5341 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5401 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5461 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     5521 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     5581 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     5641 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     5701 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     5761 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5821 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5881 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5941 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6001 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6061 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6121 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6181 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6241 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6301 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6361 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6421 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6481 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6541 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6601 ccatgcat
//
