LOCUS       sequence-383731-        8524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mVenus tagged human kinesin-1 (KIF5B aa1-560) to 
            anchor microtubules.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8524)
  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 8524)
  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(1-560)Rigor-L-mVenus
FEATURES             Location/Qualifiers
     source          1..8524
                     /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             join(2811..2813,2814..3527)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2811..2813,2814..3527)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2853..2874)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3114..3133)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3461..3482
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3682..3816
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3719..3738)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3773..3792
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3830..3846)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3830..3846)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3843..3865)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3854..3870
                     /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(3878..3884,3885..3902,3903..3908))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3923..3944
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4061..4078)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4232..4820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4312..4331)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4991..5782,5783..5851))
                     /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     5614..5633
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5852..5956)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6024..6042
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6080..6102)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6202..6221
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6415..6437
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6429..6446
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6430..6446
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8247..8522
                     /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 gggccgccyc gggccggggr gggctcgggg grggggcgcg 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 gctagtgaaa
     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 ctgcggaaca
     2101 ctattcagtg gcttgaaaat gagctcaaca gatggcgtaa tggggagacg gtgcctattg
     2161 atgaacagtt tgacaaagag aaagccaact tggaagcttt cacagtggat aaagatatta
     2221 ctcttaccaa tgataaacca gcaaccgcaa ttggagttat aggaaatttt actgatgctg
     2281 aaagaagaaa gtgtgaagaa gaaattgcta aattatacaa acagcttgat gacaaggatg
     2341 aagaaattaa ccagcaaagt caactggtag agaaactgaa gacgcaaatg ttggatcagg
     2401 aggagctttt ggcatctacc agaagggatc aagacaatat gcaagctgag ctgaatcgcc
     2461 ttcaagcaga aaatgatgcc tctaaagaag aagtgaaaga agttttacag gccctagaag
     2521 aacttgctgt caattatgat cagaagtctc aggaagttga agacaaaact aaggaatatg
     2581 aattgcttag tgatgaattg aatcagaaat cggcaacttt agcgagtata gatgctgagc
     2641 ttcagaaact taaggaaatg accaaccacc agaaaaaacg agcagctgag atgatggcat
     2701 ctttactaaa atctagaggt ggcgggggca gtgggggagg gggaagtgga ggcggcggtt
     2761 ctggcggggg aggctccggt ggcgggggga gcgggggtgg gagtaagctt gtgagcaagg
     2821 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     2881 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     2941 tgaagttgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     3001 tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     3061 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     3121 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     3181 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     3241 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggcaa
     3301 acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc
     3361 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     3421 agtccaaact gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     3481 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtgaata cgtatgtgta
     3541 attatgtgaa cgcgtaccgg tcctagataa gtaatgatca taatcagcca tatcacatct
     3601 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     3661 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     3721 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3781 tccaaactca tcaatgtatc ttatcatgtc tggatcaatt cgtaatcatg tcatagctgt
     3841 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3901 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     3961 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4021 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4081 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4141 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4201 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4261 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4321 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4381 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4441 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4501 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4561 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4621 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4681 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4741 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4801 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4861 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4921 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4981 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5041 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5101 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5161 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5221 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5281 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5341 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5401 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5461 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5521 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5581 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5641 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5701 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5761 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5821 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5881 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5941 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     6001 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     6061 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     6121 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     6181 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     6241 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     6301 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     6361 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     6421 cacgacgttg taaaacgacg gccagtgcca agttgggatc tttgcattgg cccacggctc
     6481 tcaggatggg gatgctcccc ttcagcaccc ggttcccctt ggaaactgat ggtcctggct
     6541 ctgtggcatg gcagtggcac tgtgaggagc cccctaccag cagcacacag tggggttggc
     6601 actgccacgc tccggatgcc gcgctctgat ccaaccccat aatcaaggga acccgaattg
     6661 ccccatcatt gcccccacca cccccatcct gccgggccct cacaccccac gctgccttct
     6721 ggtgacattc cccagcccaa acccacggct tcatggctac cgcggggcat ttcccattgc
     6781 cgccccatta tcagctctgc acacctcccg ctgtacccat gcctcgtggc tgcccttctt
     6841 tgacgtataa tcttctaatt aatacccggc cttgtcaaag tggagcacaa acgttaatta
     6901 attccccagc aggcaggtaa ttaacagtgt gactcccttt ttgctgcgag tggggctgat
     6961 acagagagat gtggcactat ggagcccacg gggtcctggc actgggtgcc cacggaggtc
     7021 cccatgtgct gcagtgtcac cgcctccgag gagacagtat tgtccctgcg gtgtccctgc
     7081 agctcagctc tgtccacagg gccacctcca gtttggaggg gacacaatgc agccccgatg
     7141 caacccatcc tcgcagcatc ccagggacaa agaccccact gcaagaccgc acacagggct
     7201 gggtcccgct cccctaatat ctacagtgct tcttgcatgg ccccttaatc aatgcagtta
     7261 atcagcatgc gctcatgcac cgctctggag ctgcaaagcc cctcgcagcg ctgctcacca
     7321 acaccgcgca ccgccccggc ccagcctgca gcacgcgctg caaacaggaa agaaacaaaa
     7381 tattgcccaa atgtaggcaa aggcattcgg ctgccttgac ctccgccggg ccgggccctg
     7441 cctgactcag ctccttactc agcgctcgct tcctccctcc ggctgccacc gccgcagcgc
     7501 acaccctgac aaagagtggc ccttaacggg ctctgaggtg cacccagcag tgcactcagc
     7561 agtccaaggg ccggcctgga ggtttgcacc gctacgtgct gacattagca ttgaacttgg
     7621 ccctgggtag tgctgcaggc cgggcggggt gggtgtagag agtgcagcgc gcgttgcacc
     7681 cggtgcccct tcccctccct tgcatcccag caggctgcac cccagcacca ggcccgtgca
     7741 tgcatgctcc tggtgttatt gcagcctggt gcatgcatgc gtcttgatgg tgcagcgctg
     7801 tgcatgcatc ctccttggtg tgtagcagtc tagtgcatgc atacccctcg gtgttattgc
     7861 tgctctgtgc acgcacgctc attgtatcac ttcatcccag tgcatgcact cacactggag
     7921 cgattgctgc tcggtgcacg cacactcatt gtatcactgc agctcagtgg ctgcacgcac
     7981 accggtgtta ttgctgctcg gtgcgtgcat gcacatcagt gtcgctgcag ctcagtgcat
     8041 gcacgctcat tgcccatcgc tatccctgcc tctcctgctg gcgctccccg ggaggtgact
     8101 tcaaggggac cgcaggacca cctcgggggt ggggggaggg ctgcacacgc ggaccccgct
     8161 ccccctcccc aacaaagcac tgtggaatca aaaagggggg aggggggatg gaggggcgcg
     8221 tcacaccccc gccccacacc ctcacctcga ggtgagcccc acgttctgct tcactctccc
     8281 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
     8341 agcgatgggg gcgggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgrggggc
     8401 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
     8461 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
     8521 cggg
//
