LOCUS       sequence-383793-        7977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of dimeric, GFP tagged, human kinesin-1 (KIF5B aa1-353).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7977)
  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 7977)
  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-GFP
FEATURES             Location/Qualifiers
     source          1..7977
                     /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(2277..2279,2280..2993)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2277..2279,2280..2990)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2319..2340)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2580..2599)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2927..2948
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3135..3269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3172..3191)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3226..3245
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3283..3299)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3283..3299)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3296..3318)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3307..3323
                     /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(3331..3337,3338..3355,3356..3361))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3376..3397
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3514..3531)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3685..4273)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3765..3784)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4444..5235,5236..5304))
                     /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     5067..5086
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5305..5409)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5477..5495
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5533..5555)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5655..5674
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5868..5890
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5882..5899
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5883..5899
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7700..7975
                     /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 kgggggkgcc gggcggggcg
      601 gggccgccyc gggccggggg 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 cttgaagaca
     2101 agatcgaaga actactttcg aaaatctacc accttgaaaa tgagatcgcc agacttaaga
     2161 aattaatcgg cgaaatttct agaggtggcg ggggcagtgg gggaggggga agtggaggcg
     2221 gcggttctgg cgggggaggc tccggtggcg gggggagcgg gggtgggagt aagcttgtga
     2281 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     2341 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     2401 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     2461 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     2521 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     2581 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     2641 gcattgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     2701 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     2761 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     2821 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     2881 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     2941 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag tgaactagtt
     3001 gaacgcgtac cggtcctaga taagtaatga tcataatcag ccatatcaca tctgtagagg
     3061 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3121 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3181 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3241 tcatcaatgt atcttatcat gtctggatca attcgtaatc atgtcatagc tgtttcctgt
     3301 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3361 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3421 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3481 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3541 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3601 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3661 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3721 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3781 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3841 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3901 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3961 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4021 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4081 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4141 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4201 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4261 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4321 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4381 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4441 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4501 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4561 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4621 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4681 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4741 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4801 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4861 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4921 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4981 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5041 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5101 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5161 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5221 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5281 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5341 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5401 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     5461 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     5521 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     5581 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     5641 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     5701 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     5761 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     5821 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     5881 ttgtaaaacg acggccagtg ccaagttggg atctttgcat tggcccacgg ctctcaggat
     5941 ggggatgctc cccttcagca cccggttccc cttggaaact gatggtcctg gctctgtggc
     6001 atggcagtgg cactgtgagg agccccctac cagcagcaca cagtggggtt ggcactgcca
     6061 cgctccggat gccgcgctct gatccaaccc cataatcaag ggaacccgaa ttgccccatc
     6121 attgccccca ccacccccat cctgccgggc cctcacaccc cacgctgcct tctggtgaca
     6181 ttccccagcc caaacccacg gcttcatggc taccgcgggg catttcccat tgccgcccca
     6241 ttatcagctc tgcacacctc ccgctgtacc catgcctcgt ggctgccctt ctttgacgta
     6301 taatcttcta attaataccc ggccttgtca aagtggagca caaacgttaa ttaattcccc
     6361 agcaggcagg taattaacag tgtgactccc tttttgctgc gagtggggct gatacagaga
     6421 gatgtggcac tatggagccc acggggtcct ggcactgggt gcccacggag gtccccatgt
     6481 gctgcagtgt caccgcctcc gaggagacag tattgtccct gcggtgtccc tgcagctcag
     6541 ctctgtccac agggccacct ccagtttgga ggggacacaa tgcagccccg atgcaaccca
     6601 tcctcgcagc atcccaggga caaagacccc actgcaagac cgcacacagg gctgggtccc
     6661 gctcccctaa tatctacagt gcttcttgca tggcccctta atcaatgcag ttaatcagca
     6721 tgcgctcatg caccgctctg gagctgcaaa gcccctcgca gcgctgctca ccaacaccgc
     6781 gcaccgcccc ggcccagcct gcagcacgcg ctgcaaacag gaaagaaaca aaatattgcc
     6841 caaatgtagg caaaggcatt cggctgcctt gacctccgcc gggccgggcc ctgcctgact
     6901 cagctcctta ctcagcgctc gcttcctccc tccggctgcc accgccgcag cgcacaccct
     6961 gacaaagagt ggcccttaac gggctctgag gtgcacccag cagtgcactc agcagtccaa
     7021 gggccggcct ggaggtttgc accgctacgt gctgacatta gcattgaact tggccctggg
     7081 tagtgctgca ggccgggcgg ggtgggtgta gagagtgcag cgcgcgttgc acccggtgcc
     7141 ccttcccctc ccttgcatcc cagcaggctg caccccagca ccaggcccgt gcatgcatgc
     7201 tcctggtgtt attgcagcct ggtgcatgca tgcgtcttga tggtgcagcg ctgtgcatgc
     7261 atcctccttg gtgtgtagca gtctagtgca tgcatacccc tcggtgttat tgctgctctg
     7321 tgcacgcacg ctcattgtat cacttcatcc cagtgcatgc actcacactg gagcgattgc
     7381 tgctcggtgc acgcacactc attgtatcac tgcagctcag tggctgcacg cacaccggtg
     7441 ttattgctgc tcggtgcgtg catgcacatc agtgtcgctg cagctcagtg catgcacgct
     7501 cattgcccat cgctatccct gcctctcctg ctggcgctcc ccgggaggtg acttcaaggg
     7561 gaccgcagga ccacctcggg ggtgggggga gggctgcaca cgcggacccc gctccccctc
     7621 cccaacaaag cactgtggaa tcaaaaaggg gggagggggg atggaggggc gcgtcacacc
     7681 cccgccccac accctcacct cgaggtgagc cccacgttct gcttcactct ccccatctcc
     7741 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
     7801 ggggcggggg ggggggggnc gcgcgccagg cggggcgggg cggggcgrgg ggcggggcgg
     7861 ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
     7921 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcggg
//
