LOCUS       sequence-100642-        5302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Non-fluorescent, truncated kinesin-1 fused to split GFP linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5302)
  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 5302)
  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)-GFP11
FEATURES             Location/Qualifiers
     source          1..5302
                     /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"
     CDS             1928..1975
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     polyA_signal    2089..2210
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2126..2145)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2180..2199
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2217..2672)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2354..2375)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2566..2585
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2699..2803
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2805..3162
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2805..2996
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2828..2848)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3013..3148
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3197..3991
                     /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             3197..3991
                     /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(3251..3270)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3861..3880
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4179..4198)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4223..4270
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4599..5187
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5088..5107
                     /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 gaccggtagg
     1921 cagcggcaga gaccacatgg tgctgcacga gtacgtgaac gccgccggca tcacctgatc
     1981 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2041 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2101 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2161 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     2221 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2281 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     2341 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2401 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     2461 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     2521 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     2581 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     2641 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     2701 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2761 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     2821 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2881 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2941 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3001 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3061 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3121 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     3181 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3241 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3301 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3361 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3421 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3481 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3541 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3601 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3661 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     3721 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3781 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     3841 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     3901 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     3961 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4021 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4081 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4141 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     4201 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     4261 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     4321 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     4381 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     4441 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4501 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4561 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4621 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4681 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4741 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4801 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4861 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4921 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4981 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5041 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5101 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5161 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5221 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5281 aaccgtatta ccgccatgca tc
//
