LOCUS       sequence-100656-        6879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tandem mCherry-labeled, forced-dimeric, truncated kinesin-3 fused to
            EF Hand linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6879)
  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 6879)
  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: KIF1A(1-393)-GCN4-2xmCherry-EF(C)
FEATURES             Location/Qualifiers
     source          1..6879
                     /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"
     regulatory      644..653
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1626..1635
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1967..1976
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3667..3788
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3704..3723)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3758..3777
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3795..4250)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3932..3953)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4144..4163
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4277..4381
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4383..4740
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4383..4574
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4406..4426)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4591..4726
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4775..5569
                     /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             4775..5569
                     /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(4829..4848)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5439..5458
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5757..5776)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5801..5848
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6177..6765
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6666..6685
                     /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 attcgggaag cttgccacca tggctggggc
      661 ctctgtgaag gtggcggtgc gcgtccgccc cttcaactcc cgggaaatga gccgcgactc
      721 caagtgcatc attcagatgt ctggaaacac caccaccatt gtcaacccca aacagcccaa
      781 ggagacaccc aagagcttca gcttcgatta ctcctactgg tcacatacct cgcccgagga
      841 catcaactat gcctctcaga agcaggtcta cagggacatt ggggaggaga tgctacagca
      901 tgcctttgag ggctacaacg tgtgcatctt tgcctatgga cagacagggg ctggcaagtc
      961 ctacaccatg atgggcaaac aagagaagga ccagcaaggc atcatcccac agctctgtga
     1021 agacctcttc tctcgaatca atgacacgac caacgacaac atgtcctatt ctgtggaggt
     1081 cagctacatg gagatttact gtgagcgagt ccgcgacctc ctgaacccca agaacaaagg
     1141 caacctgcgt gtgagggagc accctctgct ggggccttat gtggaggacc tctccaagct
     1201 ggctgtgacc tcctacaatg acatccagga cctcatggac tcagggaaca aggccaggac
     1261 tgtggcggcc acaaacatga atgagaccag cagccgttct cacgccgtct tcaacatcat
     1321 cttcacccag aagcgccatg atgcggagac caacatcacc acagagaaag tgagcaaaat
     1381 cagcctggtg gacctggcgg gcagtgaacg agccgactcc acgggagcca agggcacacg
     1441 actcaaggag ggagccaaca tcaacaagtc tttgaccacc ctgggaaagg tcatctctgc
     1501 tctcgctgaa atggactctg gacccaataa gaacaagaaa aagaagaaga cagacttcat
     1561 cccttaccga gactcggtat tgacatggct tctccgggaa aacctgggcg gcaactcaag
     1621 gacagccatg gtggcagccc tgagccctgc ggacatcaac tatgatgaga ccctcagcac
     1681 actgagatat gcagaccggg ccaaacagat ccgctgcaac gccattatca atgaggaccc
     1741 taacaacaag ctgattcgag aactgaagga tgaggtgacc aggctgcgag atctgctgta
     1801 cgcccaaggt cttggcgata tcactgacgg cgcgggagtg aaacaacttg aagacaaggt
     1861 tgaagaattg gcttcgaaaa attatcactt ggaaaatgag gttgccagat taaagaaatt
     1921 agttggggta ccgggaggtg gtggaggagg aggtggagga ccggtcgcca ccatggtcag
     1981 caaaggggaa gaggataaca tggccatcat caaagagttc atgcggttca aggtgcacat
     2041 ggaaggcagc gtgaacggac atgagttcga gatcgaagga gaaggagagg gacgccctta
     2101 cgagggaact cagaccgcaa agctgaaagt gactaagggt ggccctctgc cgtttgcgtg
     2161 ggacattctg tccccgcaat tcatgtatgg ctcaaaagcc tacgtcaaac acccagctga
     2221 catcccagac tacctgaaac tcagctttcc ggaaggattc aagtgggaac gcgtcatgaa
     2281 tttcgaagat ggcggagtgg tcaccgtgac tcaagacagc tcgttgcaag acggggagtt
     2341 tatctacaag gtgaagctcc ggggaactaa cttcccatca gacggtccgg tcatgcagaa
     2401 aaagacgatg ggttgggaag cctcctcgga gagaatgtac ccagaggacg gcgcgctgaa
     2461 gggtgaaatc aaacagagac tcaaactgaa agatggagga cactacgacg cggaagtcaa
     2521 gaccacctac aaggcaaaga aaccagtgca gttgccagga gcgtacaacg tgaacatcaa
     2581 acttgatatc acctcgcaca acgaagatta caccattgtg gagcaatacg aacgggcgga
     2641 aggaaggcat tcgaccgggg gaatggacga attatacaag ggctccggga gcggaatggt
     2701 gagcaaggga gaagaagata acatggctat tatcaaggaa tttatgcgct tcaaggttca
     2761 tatggagggc tcagtgaacg ggcacgaatt tgaaatcgag ggagagggag aaggcagacc
     2821 gtacgaaggc acccagactg ctaagctgaa ggttactaag ggaggacctc tgcccttcgc
     2881 atgggacatc ctttcgccgc agttcatgta cggttccaag gcttatgtga agcacccggc
     2941 agacattccg gattacctga agctgtcatt ccctgagggc tttaagtggg agcgcgtgat
     3001 gaactttgaa gatggtggag tggtgactgt gacccaagac tcctccctgc aagatgggga
     3061 gttcatctac aaagtcaagt tgcgcggaac taatttccct tcggacgggc cggtgatgca
     3121 gaagaaaact atgggatggg aagcttcatc ggaaaggatg taccccgagg acggagccct
     3181 gaaaggggag atcaagcagc ggctcaagct caaggatggc ggtcattacg acgccgaagt
     3241 caagaccacg tacaaggcca agaagccggt gcagctccct ggcgcgtaca atgtcaatat
     3301 caagctcgac atcacgtccc ataatgagga ttatactatc gtggaacagt acgagcgcgc
     3361 cgaaggacgg cactcaaccg gaggaatgga cgagctgtac aaaggttcag gaccggtttc
     3421 gggatcggga gggagcggag gatccggggg ttccggttcg acgctcgacg aacttttcga
     3481 ggaactggac aagaacggcg atggggaagt cagcttcgag gagtttcaag tgttggtaaa
     3541 gaaaatctcc cagtgatcta gatcataatc agccatacca catttgtaga ggttttactt
     3601 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3661 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3721 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3781 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3841 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3901 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3961 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     4021 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     4081 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     4141 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     4201 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     4261 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4321 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4381 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4441 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4501 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4561 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4621 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4681 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4741 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4801 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4861 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4921 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4981 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5041 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5101 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5161 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5221 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5281 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5341 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5401 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5461 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5521 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5581 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5641 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5701 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     5761 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5821 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5881 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5941 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     6001 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     6061 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6121 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6181 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6241 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6301 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     6361 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6421 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6481 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6541 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6601 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6661 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6721 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6781 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6841 tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
