LOCUS       sequence-368400-        6598 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. The delta6 mutation 
            removes the first 6 amino acids..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6598)
  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 6598)
  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,Delta-6)-Halo-Flag
FEATURES             Location/Qualifiers
     source          1..6598
                     /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             1503..1514
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     regulatory      2332..2341
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3238..3261
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3386..3507
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3423..3442)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3477..3496
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3514..3969)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3651..3672)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3863..3882
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3996..4100
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4102..4459
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4102..4293
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4125..4145)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4310..4445
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4494..5288
                     /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             4494..5288
                     /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(4548..4567)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5158..5177
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5476..5495)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5520..5567
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5896..6484
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6385..6404
                     /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 attcgcacca tgtgcagcat caaagtgatg
      661 tgccggttcc ggcccctcaa cgaagctgag atcctccgcg gggacaaatt catccccaaa
      721 ttcaaaggcg aggagacggt ggtgatcggg caagggaagc cgtatgtctt tgaccgagtt
      781 ctgccgccca acacaaccca ggagcaggtc tacaatgcct gtgcgaagca gattgtcaaa
      841 gatgtcctcg agggttacaa tgggacaatt tttgcatacg ggcagacttc atcaggaaaa
      901 actcatacca tggaggggaa gctgcacgat cctcagctta tgggtatcat acccaggatc
      961 gcacacgata ttttcgatca catctattcc atggatgaga acttggagtt tcatatcaag
     1021 gtttcctatt ttgagatcta cttggacaaa ataagggact tgcttgacgt gtccaagacc
     1081 aacttggctg ttcatgaaga caaaaacaga gtcccctatg taaaggggtg caccgagagg
     1141 tttgtgtcaa gcccggagga ggtcatggat gtgatcgatg agggcaaagc aaaccgacat
     1201 gtggctgtga caaacatgaa cgaacacagc tccaggagtc acagtatctt cctgattaac
     1261 attaaacaag agaatgtgga gactgaaaaa aaactcagcg ggaagctgta tttggttgat
     1321 ttggctggga gtgaaaaggt cagcaaaact ggtgccgagg gagctgttct tgatgaagct
     1381 aaaaatatca acaagtcttt gtctgctctt ggaaatgtga tctctgcctt ggcagaaggg
     1441 acaaaaacac atgtgccata ccgggacagc aagatgactc ggattctcca ggactctctg
     1501 ggtgggaact gcaggaccac cattgtcatt tgctgttctc cttcagtctt caatgaggct
     1561 gagaccaagt ccacgctgat gtttggacag agagcaaaga ccatcaagaa tacagtctct
     1621 gtgaacttgg aactaacagc agaagaatgg aagaagaaat atgaaaaaga gaaagagaag
     1681 aataaggccc tgaagagtgt catccagcat ctggaggtgg agctgaacag gtggaggaac
     1741 ggggaagctg tgcctgagga tgaacagatc agtgccaagg accagaagaa cctggagccc
     1801 tgcgacaaca cacccatcat agacaacatc acacctgtgg ttgacggcat ctctgctgag
     1861 aaagagaagt atgacgagga aatcaccagt ctgtaccgac agctggatga caaggacgat
     1921 gaaattaacc agcagagcca gctggctgaa aagctgaagc aacagatgtt ggatcaggac
     1981 gagcttctag cttccaccag aagggactac gagaaaatcc aggaggagct gacccgcctc
     2041 cagattgaaa acgaggcggc taaagacgag gtgaaggagg tcctccaggc cctggaggag
     2101 ctggctgtca attacgacca gaagtcacag gaagtggagg acaagacccg ggccaacgag
     2161 caactgaccg atgagctggc ccagaaaacg acgacactga caaccaccca gcgggagctg
     2221 agtcagctac aggagcttag caaccaccag aagaagaggg ccacggagat cctgaacctg
     2281 cttctcaagg atctgggcga gctggtaccg cgggcccggg atccaccggt cgccaccatg
     2341 ggttccgaaa tcggtacagg cttccccttc gacccccatt atgtggaagt cctgggcgag
     2401 cgtatgcact acgtcgatgt tggaccgcgg gatggcacgc ctgtgctgtt cctgcacggt
     2461 aacccgacct cgtcctacct gtggcgcaac atcatcccgc atgtagcacc gagtcatcgg
     2521 tgcattgctc cagacctgat cgggatggga aaatcggaca aaccagacct cgattatttc
     2581 ttcgacgacc acgtccgcta cctcgatgcc ttcatcgaag ccttgggttt ggaagaggtc
     2641 gtcctggtca tccacgactg gggctcagct ctcggattcc actgggccaa gcgcaatccg
     2701 gaacgggtca aaggtattgc atgtatggaa ttcatccggc ctatcccgac gtgggacgaa
     2761 tggccagaat tcgcccgtga gaccttccag gccttccgga ccgccgacgt cggccgagag
     2821 ttgatcatcg atcagaacgc tttcatcgag ggtgcgctcc cgatgggggt cgtccgtccg
     2881 cttacggagg tcgagatgga ccactatcgc gagcccttcc tcaagcctgt tgaccgagag
     2941 ccactgtggc gattccccaa cgagctgccc atcgccggtg agcccgcgaa catcgtcgcg
     3001 ctcgtcgagg catacatgaa ctggctgcac cagtcacctg tcccgaagtt gttgttctgg
     3061 ggcacacccg gcgtactgat ccccccggcc gaagccgcga gacttgccga aagcctcccc
     3121 aactgcaaga cagtggacat cggcccggga ttgttcttgc tccaggaaga caacccggac
     3181 cttatcggca gtgagatcgc gcgctggctc ccaggactgg ccggcggtgt acaaggagat
     3241 tacaaggatg acgatgacaa gtgagcggcc gcgactctag atcataatca gccataccac
     3301 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     3361 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     3421 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3481 tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa
     3541 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     3601 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     3661 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     3721 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     3781 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3841 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     3901 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3961 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4021 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4081 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     4141 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4201 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4261 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4321 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4381 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4441 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     4501 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4561 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4621 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     4681 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4741 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4801 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4861 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4921 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4981 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     5041 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5101 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5161 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5221 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5281 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5341 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5401 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5461 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     5521 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     5581 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     5641 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     5701 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     5761 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5821 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5881 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5941 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6001 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6061 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6121 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6181 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6241 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6301 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6361 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6421 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6481 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6541 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
