LOCUS       sequence-100644-        7266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tandem mCherry-labeled, truncated kinesin-1 fused to EF Hand linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7266)
  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 7266)
  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)-2xmCherry-EF(C)
FEATURES             Location/Qualifiers
     source          1..7266
                     /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      2354..2363
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4054..4175
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4091..4110)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4145..4164
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4182..4637)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4319..4340)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4531..4550
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4664..4768
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4770..5127
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4770..4961
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4793..4813)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4978..5113
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5162..5956
                     /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             5162..5956
                     /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(5216..5235)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5826..5845
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6144..6163)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6188..6235
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6564..7152
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7053..7072
                     /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 ggaggtggtg gaggaggagg
     2341 tggaggaccg gtcgccacca tggtcagcaa aggggaagag gataacatgg ccatcatcaa
     2401 agagttcatg cggttcaagg tgcacatgga aggcagcgtg aacggacatg agttcgagat
     2461 cgaaggagaa ggagagggac gcccttacga gggaactcag accgcaaagc tgaaagtgac
     2521 taagggtggc cctctgccgt ttgcgtggga cattctgtcc ccgcaattca tgtatggctc
     2581 aaaagcctac gtcaaacacc cagctgacat cccagactac ctgaaactca gctttccgga
     2641 aggattcaag tgggaacgcg tcatgaattt cgaagatggc ggagtggtca ccgtgactca
     2701 agacagctcg ttgcaagacg gggagtttat ctacaaggtg aagctccggg gaactaactt
     2761 cccatcagac ggtccggtca tgcagaaaaa gacgatgggt tgggaagcct cctcggagag
     2821 aatgtaccca gaggacggcg cgctgaaggg tgaaatcaaa cagagactca aactgaaaga
     2881 tggaggacac tacgacgcgg aagtcaagac cacctacaag gcaaagaaac cagtgcagtt
     2941 gccaggagcg tacaacgtga acatcaaact tgatatcacc tcgcacaacg aagattacac
     3001 cattgtggag caatacgaac gggcggaagg aaggcattcg accgggggaa tggacgaatt
     3061 atacaagggc tccgggagcg gaatggtgag caagggagaa gaagataaca tggctattat
     3121 caaggaattt atgcgcttca aggttcatat ggagggctca gtgaacgggc acgaatttga
     3181 aatcgaggga gagggagaag gcagaccgta cgaaggcacc cagactgcta agctgaaggt
     3241 tactaaggga ggacctctgc ccttcgcatg ggacatcctt tcgccgcagt tcatgtacgg
     3301 ttccaaggct tatgtgaagc acccggcaga cattccggat tacctgaagc tgtcattccc
     3361 tgagggcttt aagtgggagc gcgtgatgaa ctttgaagat ggtggagtgg tgactgtgac
     3421 ccaagactcc tccctgcaag atggggagtt catctacaaa gtcaagttgc gcggaactaa
     3481 tttcccttcg gacgggccgg tgatgcagaa gaaaactatg ggatgggaag cttcatcgga
     3541 aaggatgtac cccgaggacg gagccctgaa aggggagatc aagcagcggc tcaagctcaa
     3601 ggatggcggt cattacgacg ccgaagtcaa gaccacgtac aaggccaaga agccggtgca
     3661 gctccctggc gcgtacaatg tcaatatcaa gctcgacatc acgtcccata atgaggatta
     3721 tactatcgtg gaacagtacg agcgcgccga aggacggcac tcaaccggag gaatggacga
     3781 gctgtacaaa ggttcaggac cggtttcggg atcgggaggg agcggaggat ccgggggttc
     3841 cggttcgacg ctcgacgaac ttttcgagga actggacaag aacggcgatg gggaagtcag
     3901 cttcgaggag tttcaagtgt tggtaaagaa aatctcccag tgatctagat cataatcagc
     3961 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     4021 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     4081 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4141 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     4201 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4261 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4321 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4381 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4441 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4501 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4561 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4621 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4681 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4741 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     4801 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4861 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4921 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4981 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5041 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     5101 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     5161 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     5221 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5281 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5341 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5401 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5461 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5521 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5581 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5641 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     5701 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5761 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5821 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5881 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5941 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     6001 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     6061 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     6121 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     6181 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     6241 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     6301 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     6361 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     6421 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6481 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6541 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6601 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6661 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     6721 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6781 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6841 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6901 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6961 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7021 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7081 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7141 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7201 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7261 atgcat
//
