LOCUS       sequence-100643-        6784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tandem mCitrine-labeled, truncated kinesin-1 fused to split GFP 
            linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6784)
  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 6784)
  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)-2xmCitrine-GFP11
FEATURES             Location/Qualifiers
     source          1..6784
                     /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             3410..3457
                     /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    3571..3692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3608..3627)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3662..3681
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3699..4154)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3836..3857)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4048..4067
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4181..4285
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4287..4644
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4287..4478
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4310..4330)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4495..4630
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4679..5473
                     /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             4679..5473
                     /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(4733..4752)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5343..5362
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5661..5680)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5705..5752
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6081..6669
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6570..6589
                     /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 gaccggtcgc
     1921 catggtgtcc aagggagagg aactgtttac cggagtggtc cccatactgg tggagctcga
     1981 cggagacgtg aacggacaca aattctccgt gtctggggaa ggcgagggcg acgccacata
     2041 cggtaaattg accctcaagt ttatctgtac cacaggtaaa ctgcctgtgc cttggcctac
     2101 cctggtgact acctttggct atggcctcat gtgtttcgca cgctatcccg atcacatgaa
     2161 gcaacacgat tttttcaaaa gcgccatgcc agagggctat gtccaagagc ggaccatttt
     2221 ctttaaggat gacgggaact ataagacacg cgcagaggtc aagtttgagg gggacacact
     2281 ggtcaaccgg atcgaactga aaggcattga ctttaaggaa gatggaaaca ttctgggcca
     2341 caaactggaa tacaattaca atagccataa cgtgtatatt atggcagaca aacagaaaaa
     2401 cgggatcaaa gtgaatttca aaatccggca caatatcgaa gacgggagcg tccagctggc
     2461 cgatcactat cagcaaaaca cacccattgg cgatggccct gtgctcctgc cagacaatca
     2521 ctatctgtcc taccaatcca agctgagtaa agatcccaat gagaaacgcg accacatggt
     2581 cctcctggaa ttcgtcaccg cagccggcat caccctcggg atggatgagt tgtataaggg
     2641 cgcgccaggt actagtggta tggtcagcaa aggggaggag ctcttcacag gggtcgtgcc
     2701 tatcctcgtc gaactggatg gcgatgtaaa tggccataag tttagcgtct ccggagaggg
     2761 tgaaggggat gcaacctatg gcaaactgac tctgaaattc atatgcacaa ccgggaagct
     2821 ccccgtcccc tggcccacac tcgtcaccac attcgggtac gggctgatgt gctttgcccg
     2881 ttaccctgac catatgaaac agcatgactt ttttaaatcc gcaatgcctg aagggtacgt
     2941 gcaggaacgc acaatcttct tcaaggacga tggcaattac aaaacccggg ccgaggtgaa
     3001 attcgaaggc gataccctcg tgaatcgcat agagctcaag gggatcgatt tcaaagaaga
     3061 tggcaatatc ctcggacata agctcgagta taactataac agtcacaatg tctacatcat
     3121 ggccgataag caaaagaatg gcattaaggt caactttaag atacgccata acattgagga
     3181 tggcagtgtg caactcgcag accattacca acagaatacc ccaatcgggg acggtccagt
     3241 cctgctcccc gataaccatt acctcagcta tcagagcaaa ctcagcaagg accctaacga
     3301 aaagcgggat catatggtgc tgcttgagtt tgtgacagcc gcagggatta ctctgggcat
     3361 ggacgagctg tacaagggcg cgccaggagg ttcaccggta ggcagcggca gagaccacat
     3421 ggtgctgcac gagtacgtga acgccgccgg catcacctga tctagatcat aatcagccat
     3481 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     3541 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3601 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3661 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt
     3721 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3781 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     3841 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3901 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     3961 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4021 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4081 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4141 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4201 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4261 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     4321 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4381 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4441 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4501 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4561 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4621 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     4681 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4741 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4801 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4861 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4921 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4981 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5041 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5101 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5161 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     5221 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5281 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5341 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5401 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5461 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5521 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5581 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5641 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5701 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     5761 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     5821 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     5881 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     5941 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6001 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6061 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6121 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6181 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     6241 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6301 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6361 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6421 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6481 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6541 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6601 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6661 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6721 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     6781 catc
//
