LOCUS       sequence-100640-        5963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Non-fluorescent, truncated kinesin-1 fused to DmrC linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5963)
  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 5963)
  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)-DmrC
FEATURES             Location/Qualifiers
     source          1..5963
                     /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             1506..1517
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             2362..2640
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     polyA_signal    2751..2872
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2788..2807)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2842..2861
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2879..3334)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3016..3037)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3228..3247
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3361..3465
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3467..3824
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3467..3658
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3490..3510)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3675..3810
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3859..4653
                     /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             3859..4653
                     /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(3913..3932)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4523..4542
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4841..4860)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4885..4932
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5261..5849
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5750..5769
                     /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 caagcttatg gcggatccag ccgaatgcag catcaaagtg
      661 atgtgccggt tccggcccct caacgaagct gagatcctcc gcggggacaa attcatcccc
      721 aaattcaaag gcgaggagac ggtggtgatc gggcaaggga agccgtatgt ctttgaccga
      781 gttctgccgc ccaacacaac ccaggagcag gtctacaatg cctgtgcgaa gcagattgtc
      841 aaagatgtcc tcgagggtta caatgggaca atttttgcat acgggcagac ttcatcagga
      901 aaaactcata ccatggaggg gaagctgcac gatcctcagc ttatgggtat catacccagg
      961 atcgcacacg atattttcga tcacatctat tccatggatg agaacttgga gtttcatatc
     1021 aaggtttcct attttgagat ctacttggac aaaataaggg acttgcttga cgtgtccaag
     1081 accaacttgg ctgttcatga agacaaaaac agagtcccct atgtaaaggg gtgcaccgag
     1141 aggtttgtgt caagcccgga ggaggtcatg gatgtgatcg atgagggcaa agcaaaccga
     1201 catgtggctg tgacaaacat gaacgaacac agctccagga gtcacagtat cttcctgatt
     1261 aacattaaac aagagaatgt ggagactgaa aaaaaactca gcgggaagct gtatttggtt
     1321 gatttggctg ggagtgaaaa ggtcagcaaa actggtgccg agggagctgt tcttgatgaa
     1381 gctaaaaata tcaacaagtc tttgtctgct cttggaaatg tgatctctgc cttggcagaa
     1441 gggacaaaaa cacatgtgcc ataccgggac agcaagatga ctcggattct ccaggactct
     1501 ctgggtggga actgcaggac caccattgtc atttgctgtt ctccttcagt cttcaatgag
     1561 gctgagacca agtccacgct gatgtttgga cagagagcaa agaccatcaa gaatacagtc
     1621 tctgtgaact tggaactaac agcagaagaa tggaagaaga aatatgaaaa agagaaagag
     1681 aagaataagg ccctgaagag tgtcatccag catctggagg tggagctgaa caggtggagg
     1741 aacggggaag ctgtgcctga ggatgaacag atcagtgcca aggaccagaa gaacctggag
     1801 ccctgcgaca acacacccat catagacaac atcacacctg tggttgacgg catctctgct
     1861 gagaaagaga agtatgacga ggaaatcacc agtctgtacc gacagctgga tgacaaggac
     1921 gatgaaatta accagcagag ccagctggct gaaaagctga agcaacagat gttggatcag
     1981 gacgagcttc tagcttccac cagaagggac tacgagaaaa tccaggagga gctgacccgc
     2041 ctccagattg aaaacgaggc ggctaaagac gaggtgaagg aggtcctcca ggccctggag
     2101 gagctggctg tcaattacga ccagaagtca caggaagtgg aggacaagac ccgggccaac
     2161 gagcaactga ccgatgagct ggcccagaaa acgacgacac tgacaaccac ccagcgggag
     2221 ctgagtcagc tacaggagct tagcaaccac cagaagaaga gggccacgga gatcctgaac
     2281 ctgcttctca aggatctggg cgagctggta ccgggaggtg gtggaggagg aggtggagga
     2341 ccggtgggaa gcggatctag aatcctctgg catgagatgt ggcatgaagg cctggaagag
     2401 gcatctcgtt tgtactttgg ggaaaggaac gtgaaaggca tgtttgaggt gctggagccc
     2461 ttgcatgcta tgatggaacg gggcccccag actctgaagg aaacatcctt taatcaggcc
     2521 tatggtcgag atttaatgga ggcccaagag tggtgcagga agtacatgaa atcagggaat
     2581 gtcaaggacc tcctccaagc ctgggacctc tattatcatg tgttccgacg aatctcaaag
     2641 actagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     2701 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     2761 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2821 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa
     2881 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     2941 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3001 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     3061 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3121 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3181 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3241 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3301 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     3361 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3421 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     3481 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3541 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3601 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3661 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3721 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3781 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     3841 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3901 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3961 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4021 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     4081 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4141 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4201 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4261 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4321 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4381 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4441 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4501 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4561 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4621 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4681 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4741 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4801 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     4861 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     4921 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     4981 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     5041 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     5101 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5161 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5221 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5281 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5341 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5401 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5461 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5521 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5581 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5641 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5701 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5761 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5821 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5881 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5941 ataaccgtat taccgccatg cat
//
