LOCUS       sequence-383721-        8526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GFP tagged human kinesin-1 (KIF5B aa1-560).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8526)
  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 8526)
  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)-L-GFP
FEATURES             Location/Qualifiers
     source          1..8526
                     /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..3524)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2811..2813,2814..3524)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     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    3684..3818
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3721..3740)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3775..3794
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3832..3848)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3832..3848)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3845..3867)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3856..3872
                     /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(3880..3886,3887..3904,3905..3910))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3925..3946
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4063..4080)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4234..4822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4314..4333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4993..5784,5785..5853))
                     /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     5616..5635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5854..5958)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6026..6044
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6082..6104)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6204..6223
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6417..6439
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6431..6448
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6432..6448
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8249..8524
                     /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 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 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 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     3001 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     3061 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     3121 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcattg
     3181 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     3241 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     3301 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     3361 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     3421 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     3481 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caaggctagc ggaggcggtt
     3541 gaggctcagg cacgcgtacc ggtcctagat aagtaatgat cataatcagc catatcacat
     3601 ctgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     3661 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3721 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3781 tgtccaaact catcaatgta tcttatcatg tctggatcaa ttcgtaatca tgtcatagct
     3841 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3901 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3961 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4021 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4081 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4141 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4201 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4261 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4321 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4381 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4441 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4501 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4561 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4621 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4681 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4741 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4801 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4861 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4921 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4981 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5041 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5101 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5161 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5221 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5281 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5341 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5401 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5461 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5521 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5581 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5641 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5701 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5761 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5821 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5881 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5941 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     6001 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     6061 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     6121 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     6181 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     6241 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     6301 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     6361 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     6421 gtcacgacgt tgtaaaacga cggccagtgc caagttggga tctttgcatt ggcccacggc
     6481 tctcaggatg gggatgctcc ccttcagcac ccggttcccc ttggaaactg atggtcctgg
     6541 ctctgtggca tggcagtggc actgtgagga gccccctacc agcagcacac agtggggttg
     6601 gcactgccac gctccggatg ccgcgctctg atccaacccc ataatcaagg gaacccgaat
     6661 tgccccatca ttgcccccac cacccccatc ctgccgggcc ctcacacccc acgctgcctt
     6721 ctggtgacat tccccagccc aaacccacgg cttcatggct accgcggggc atttcccatt
     6781 gccgccccat tatcagctct gcacacctcc cgctgtaccc atgcctcgtg gctgcccttc
     6841 tttgacgtat aatcttctaa ttaatacccg gccttgtcaa agtggagcac aaacgttaat
     6901 taattcccca gcaggcaggt aattaacagt gtgactccct ttttgctgcg agtggggctg
     6961 atacagagag atgtggcact atggagccca cggggtcctg gcactgggtg cccacggagg
     7021 tccccatgtg ctgcagtgtc accgcctccg aggagacagt attgtccctg cggtgtccct
     7081 gcagctcagc tctgtccaca gggccacctc cagtttggag gggacacaat gcagccccga
     7141 tgcaacccat cctcgcagca tcccagggac aaagacccca ctgcaagacc gcacacaggg
     7201 ctgggtcccg ctcccctaat atctacagtg cttcttgcat ggccccttaa tcaatgcagt
     7261 taatcagcat gcgctcatgc accgctctgg agctgcaaag cccctcgcag cgctgctcac
     7321 caacaccgcg caccgccccg gcccagcctg cagcacgcgc tgcaaacagg aaagaaacaa
     7381 aatattgccc aaatgtaggc aaaggcattc ggctgccttg acctccgccg ggccgggccc
     7441 tgcctgactc agctccttac tcagcgctcg cttcctccct ccggctgcca ccgccgcagc
     7501 gcacaccctg acaaagagtg gcccttaacg ggctctgagg tgcacccagc agtgcactca
     7561 gcagtccaag ggccggcctg gaggtttgca ccgctacgtg ctgacattag cattgaactt
     7621 ggccctgggt agtgctgcag gccgggcggg gtgggtgtag agagtgcagc gcgcgttgca
     7681 cccggtgccc cttcccctcc cttgcatccc agcaggctgc accccagcac caggcccgtg
     7741 catgcatgct cctggtgtta ttgcagcctg gtgcatgcat gcgtcttgat ggtgcagcgc
     7801 tgtgcatgca tcctccttgg tgtgtagcag tctagtgcat gcatacccct cggtgttatt
     7861 gctgctctgt gcacgcacgc tcattgtatc acttcatccc agtgcatgca ctcacactgg
     7921 agcgattgct gctcggtgca cgcacactca ttgtatcact gcagctcagt ggctgcacgc
     7981 acaccggtgt tattgctgct cggtgcgtgc atgcacatca gtgtcgctgc agctcagtgc
     8041 atgcacgctc attgcccatc gctatccctg cctctcctgc tggcgctccc cgggaggtga
     8101 cttcaagggg accgcaggac cacctcgggg gtggggggag ggctgcacac gcggaccccg
     8161 ctccccctcc ccaacaaagc actgtggaat caaaaagggg ggagggggga tggaggggcg
     8221 cgtcacaccc ccgccccaca ccctcacctc gaggtgagcc ccacgttctg cttcactctc
     8281 cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt
     8341 gcagcgatgg gggcgggggg gggggggncg cgcgccaggc ggggcggggc ggggcgrggg
     8401 gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
     8461 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
     8521 ggcggg
//
