LOCUS       sequence-383790-        8214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rapalog-inducible coupling to dimeric human kinesin-1 (KIF5B 
            aa1-560) via FKBP for anterograde transport.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8214)
  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 8214)
  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-2xHA-L-FRB
FEATURES             Location/Qualifiers
     source          1..8214
                     /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             2811..2837
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2838..2864
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2964..3242
                     /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    3372..3506
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3409..3428)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3463..3482
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3520..3536)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3520..3536)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3533..3555)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3544..3560
                     /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(3568..3574,3575..3592,3593..3598))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3613..3634
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3751..3768)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3922..4510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4002..4021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4681..5472,5473..5541))
                     /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     5304..5323
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5542..5646)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5714..5732
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5770..5792)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5892..5911
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6105..6127
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6119..6136
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6120..6136
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7937..8212
                     /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 rggctccggt ggcgggggga gcgggggtgg gagtaagctt tacccttacg
     2821 acgtccccga ctacgcatat ccatatgatg ttccggatta cgctgctagc ggaggcggtg
     2881 gctcaggcgg aggagggtca ggagggggag gcagtggcgg gggcggctct ggaggagggg
     2941 gttctggtgg tggtagctac gtaatcctct ggcatgagat gtggcatgaa ggcctggaag
     3001 aggcatctcg tttgtacttt ggggaaagga acgtgaaagg catgtttgag gtgctggagc
     3061 ccttgcatgc tatgatggaa cggggccccc agactctgaa ggaaacatcc tttaatcagg
     3121 cctatggtcg agatttaatg gaggcccaag agtggtgcag gaagtacatg aaatcaggga
     3181 atgtcaagga cctcctccaa gcctgggacc tctattatca tgtgttccga cgaatctcaa
     3241 agtgaaccgg tcctagataa gtaatgatca taatcagcca tatcacatct gtagaggttt
     3301 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     3361 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3421 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3481 tcaatgtatc ttatcatgtc tggatcaatt cgtaatcatg tcatagctgt ttcctgtgtg
     3541 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3601 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3661 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3721 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3781 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3841 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3901 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3961 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4021 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4081 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4141 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4201 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4261 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4321 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4381 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4441 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4501 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4561 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4621 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4681 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4741 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4801 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4861 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4921 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4981 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5041 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5101 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5161 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5221 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5281 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5341 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5401 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5461 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5521 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5581 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5641 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     5701 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     5761 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     5821 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     5881 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     5941 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     6001 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     6061 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     6121 taaaacgacg gccagtgcca agttgggatc tttgcattgg cccacggctc tcaggatggg
     6181 gatgctcccc ttcagcaccc ggttcccctt ggaaactgat ggtcctggct ctgtggcatg
     6241 gcagtggcac tgtgaggagc cccctaccag cagcacacag tggggttggc actgccacgc
     6301 tccggatgcc gcgctctgat ccaaccccat aatcaaggga acccgaattg ccccatcatt
     6361 gcccccacca cccccatcct gccgggccct cacaccccac gctgccttct ggtgacattc
     6421 cccagcccaa acccacggct tcatggctac cgcggggcat ttcccattgc cgccccatta
     6481 tcagctctgc acacctcccg ctgtacccat gcctcgtggc tgcccttctt tgacgtataa
     6541 tcttctaatt aatacccggc cttgtcaaag tggagcacaa acgttaatta attccccagc
     6601 aggcaggtaa ttaacagtgt gactcccttt ttgctgcgag tggggctgat acagagagat
     6661 gtggcactat ggagcccacg gggtcctggc actgggtgcc cacggaggtc cccatgtgct
     6721 gcagtgtcac cgcctccgag gagacagtat tgtccctgcg gtgtccctgc agctcagctc
     6781 tgtccacagg gccacctcca gtttggaggg gacacaatgc agccccgatg caacccatcc
     6841 tcgcagcatc ccagggacaa agaccccact gcaagaccgc acacagggct gggtcccgct
     6901 cccctaatat ctacagtgct tcttgcatgg ccccttaatc aatgcagtta atcagcatgc
     6961 gctcatgcac cgatctggag ctgcaaagcc cctcgcagcg ctgctcacca acaccgcgca
     7021 ccgccccggc ccagcctgca gcacgcgctg caaacaggaa agaaacaaaa tattgcccaa
     7081 atgtaggcaa aggcattcgg ctgccttgac ctccgccggg ccgggccctg cctgactcag
     7141 ctccttactc agcgctcgct tcctccctcc ggctgccacc gccgcagcgc acaccctgac
     7201 aaagagtggc ccttaacggg ctctgaggtg cacccagcag tgcactcagc agtccaaggg
     7261 ccggcctgga ggtttgcacc gctacgtgct gacattagca ttgaacttgg ccctgggtag
     7321 tgctgcaggc cgggcggggt gggtgtagag agtgcagcgc gcgttgcacc cggtgcccct
     7381 tcccctccct tgcatcccag caggctgcac cccagcacca ggcccgtgca tgcatgctcc
     7441 tggtgttatt gcagcctggt gcatgcatgc gtcttgatgg tgcagcgctg tgcatgcatc
     7501 ctccttggtg tgtagcagtc tagtgcatgc atacccctcg gtgttattgc tgctctgtgc
     7561 acgcacgctc attgtatcac ttcatcccag tgcatgcact cacactggag cgattgctgc
     7621 tcggtgcacg cacactcatt gtatcactgc agctcagtgg ctgcacgcac accggtgtta
     7681 ttgctgctcg gtgcgtgcat gcacatcagt gtcgctgcag ctcagtgcat gcacgctcat
     7741 tgcccatcgc tatccctgcc tctcctgctg gcgctccccg ggaggtgact tcaaggggac
     7801 cgcaggacca cctcgggggt ggggggaggg ctgcacacgc ggaccccgct ccccctcccc
     7861 aacaaagcac tgtggaatca aaaagggggg aggggggatg gaggggcgcg tcacaccccc
     7921 gccccacacc ctcacctcga ggtgagcccc acgttctgct tcactctccc catctccccc
     7981 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
     8041 gcgggggggg gggggncgcg cgccaggcgg ggcggggcgg ggcgrggggc ggggcggggc
     8101 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
     8161 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cggg
//
