LOCUS       sequence-382800-        7968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of dimeric, mCherry tagged, human kinesin-1 (KIF5B 
            aa1-353).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7968)
  AUTHORS   Burute M, Jansen KI, Mihajlovic M, Vermonden T, Kapitein LC
  TITLE     Local changes in microtubule network mobility instruct neuronal 
            polarization and axon specification.
  JOURNAL   Sci Adv. 2022 Nov 4;8(44):eabo2343. doi: 10.1126/sciadv.abo2343. 
            Epub 2022 Nov 4.
  PUBMED    36332030
REFERENCE   2  (bases 1 to 7968)
  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"; journalName: "Sci Adv."; 
            date: "2022-11-4"; volume: "8(44):eabo2343. doi"; pages: " 
            10.1126/sciadv.abo2343. Epub 2022 Nov 4"
COMMENT     Sequence Label: pB80-hsKIF5B(1-353)-GCN4-L-mCherry
FEATURES             Location/Qualifiers
     source          1..7968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     941..963
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1026..1035
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2277..2984
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2419..2437)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    3126..3260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3163..3182)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3217..3236
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3274..3290)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3274..3290)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3287..3309)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3298..3314
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3322..3328,3329..3346,3347..3352))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3367..3388
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3505..3522)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3676..4264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3756..3775)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4435..5226,5227..5295))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5058..5077
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5296..5400)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5468..5486
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5524..5546)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5646..5665
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5859..5881
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5873..5890
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5874..5890
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7691..7966
                     /label=chicken beta-actin promoter
ORIGIN
        1 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
       61 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      121 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      181 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      241 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      301 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
      361 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
      421 gcgtgggggg gtgagcaggg ggtgtgggcg cggcggtcgg gctgtaaccc ccccctgcac
      481 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tgcggggcgt
      541 ggcgcggggc tcgccgtgcc gggcgggggg kggcggcagg kgggggtgcc gggcggggcg
      601 gggccgcctc gggccggggr gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
      661 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
      721 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
      781 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
      841 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccatc tccagcctcg gggctgtccg
      901 cagggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
      961 accggcgggg tttatatctt cccttctctg ttcctccgca gcccccaagc taggcaattg
     1021 ggcgcgccac catggcggac ctggccgagt gcaacatcaa agtgatgtgt cgcttcagac
     1081 ctctcaacga gtctgaagtg aaccgcggcg acaagtacat cgccaagttt cagggagaag
     1141 acacggtcgt gatcgcgtcc aagccttatg catttgatcg ggtgttccag tcaagcacat
     1201 ctcaagagca agtgtataat gactgtgcaa agaagattgt taaagatgta cttgaaggat
     1261 ataatggaac aatatttgca tatggacaaa catcctctgg gaagacacac acaatggagg
     1321 gtaaacttca tgatccagaa ggcatgggaa ttattccaag aatagtgcaa gatattttta
     1381 attatattta ctccatggat gaaaatttgg aatttcatat taaggtttca tattttgaaa
     1441 tatatttgga taagataagg gacctgttag atgtttcaaa gaccaacctt tcagttcatg
     1501 aagacaaaaa ccgagttccc tatgtaaagg ggtgcacaga gcgttttgta tgtagtccag
     1561 atgaagttat ggataccata gatgaaggaa aatccaacag acatgtagca gttacaaata
     1621 tgaatgaaca tagctctagg agtcacagta tatttcttat taatgtcaaa caagagaaca
     1681 cacaaacgga acaaaagctg agtggaaaac tttatctggt tgatttagct ggtagtgaaa
     1741 aggttagtaa aactggagct gaaggtgctg tgctggatga agctaaaaac atcaacaagt
     1801 cactttctgc tcttggaaat gttatttctg ctttggctga gggtagtaca tatgttccat
     1861 atcgagatag taaaatgaca agaatccttc aagattcatt aggtggcaac tgtagaacca
     1921 ctattgtaat ttgctgctct ccatcatcat acaatgagtc tgaaacaaaa tctacactct
     1981 tatttggcca aagggccaaa acaattaaga acacagtttg tgtcaatgtg gagttaactg
     2041 cagaacagtg gaaaaagaag tatgaaaaag aaaaagaaaa aaataagatc cttgaagaca
     2101 agatcgaaga actactttcg aaaatctacc accttgaaaa tgagatcgcc agacttaaga
     2161 aattaatcgg cgaaatttct agaggtggcg ggggcagtgg gggaggggga agtggaggcg
     2221 gcggttctgg cgggggaggc tccggtggcg gggggagcgg gggtgggagt aagcttgtga
     2281 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     2341 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     2401 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     2461 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     2521 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     2581 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     2641 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggtccc gtaatgcaga
     2701 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     2761 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     2821 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     2881 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     2941 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtgaactagt tgaacgcgta
     3001 ccggtcctag ataagtaatg atcataatca gccatatcac atctgtagag gttttacttg
     3061 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3121 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3181 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3241 tatcttatca tgtctggatc aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     3301 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3361 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3421 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3481 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3541 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3601 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3661 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3721 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3781 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3841 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3901 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3961 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4021 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4081 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4141 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4201 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4261 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4321 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4381 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4441 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4501 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4561 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4621 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4681 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4741 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4801 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4861 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4921 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4981 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5041 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5101 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5161 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5221 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5281 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5341 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5401 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     5461 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     5521 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     5581 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     5641 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     5701 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     5761 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     5821 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     5881 gacggccagt gccaagttgg gatctttgca ttggcccacg gctctcagga tggggatgct
     5941 ccccttcagc acccggttcc ccttggaaac tgatggtcct ggctctgtgg catggcagtg
     6001 gcactgtgag gagcccccta ccagcagcac acagtggggt tggcactgcc acgctccgga
     6061 tgccgcgctc tgatccaacc ccataatcaa gggaacccga attgccccat cattgccccc
     6121 accaccccca tcctgccggg ccctcacacc ccacgctgcc ttctggtgac attccccagc
     6181 ccaaacccac ggcttcatgg ctaccgcggg gcatttccca ttgccgcccc attatcagct
     6241 ctgcacacct cccgctgtac ccatgcctcg tggctgccct tctttgacgt ataatcttct
     6301 aattaatacc cggccttgtc aaagtggagc acaaacgtta attaattccc cagcaggcag
     6361 gtaattaaca gtgtgactcc ctttttgctg cgagtggggc tgatacagag agatgtggca
     6421 ctatggagcc cacggggtcc tggcactggg tgcccacgga ggtccccatg tgctgcagtg
     6481 tcaccgcctc cgaggagaca gtattgtccc tgcggtgtcc ctgcagctca gctctgtcca
     6541 cagggccacc tccagtttgg aggggacaca atgcagcccc gatgcaaccc atcctcgcag
     6601 catcccaggg acaaagaccc cactgcaaga ccgcacacag ggctgggtcc cgctccccta
     6661 atatctacag tgcttcttgc atggcccctt aatcaatgca gttaatcagc atgcgctcat
     6721 gcaccgctct ggagctgcaa agcccctcgc agcgctgctc accaacaccg cgcaccgccc
     6781 cggcccagcc tgcagcacgc gctgcaaaca ggaaagaaac aaaatattgc ccaaatgtag
     6841 gcaaaggcat tcggctgcct tgacctccgc cgggccgggc cctgcctgac tcagctcctt
     6901 actcagcgct cgcttcctcc ctccggctgc caccgccgca gcgcacaccc tgacaaagag
     6961 tggcccttaa cgggctctga ggtgcaccca gcagtgcact cagcagtcca agggccggcc
     7021 tggaggtttg caccgctacg tgctgacatt agcattgaac ttggccctgg gtagtgctgc
     7081 aggccgggcg gggtgggtgt agagagtgca gcgcgcgttg cacccggtgc cccttcccct
     7141 cccttgcatc ccagcaggct gcaccccagc accaggcccg tgcatgcatg ctcctggtgt
     7201 tattgcagcc tggtgcatgc atgcgtcttg atggtgcagc gctgtgcatg catcctcctt
     7261 ggtgtgtagc agtctagtgc atgcataccc ctcggtgtta ttgctgctct gtgcacgcac
     7321 gctcattgta tcacttcatc ccagtgcatg cactcacact ggagcgattg ctgctcggtg
     7381 cacgcacact cattgtatca ctgcagctca gtggctgcac gcacaccggt gttattgctg
     7441 ctcggtgcgt gcatgcacat cagtgtcgct gcagctcagt gcatgcacgc tcattgccca
     7501 tcgctatccc tgcctctcct gctggcgctc cccgggaggt gacttcaagg ggaccgcagg
     7561 accacctcgg gggtgggggg agggctgcac acgcggaccc cgctccccct ccccaacaaa
     7621 gcactgtgga atcaaaaagg ggggaggggg gatggagggg cgcgtcacac ccccgcccca
     7681 caccctcacc tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
     7741 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
     7801 gggggggggn cgcgcgccag gcggggcggg gcggggcgrg gggcggggcg gggcgaggcg
     7861 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag
     7921 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcggg
//
