LOCUS       sequence-383730-        7851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HA 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 7851)
  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 7851)
  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
FEATURES             Location/Qualifiers
     source          1..7851
                     /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"
     polyA_signal    3009..3143
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3046..3065)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3100..3119
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3157..3173)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3157..3173)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3170..3192)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3181..3197
                     /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(3205..3211,3212..3229,3230..3235))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3250..3271
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3388..3405)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3559..4147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3639..3658)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4318..5109,5110..5178))
                     /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     4941..4960
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5179..5283)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5351..5369
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5407..5429)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5529..5548
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5742..5764
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5756..5773
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5757..5773
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7574..7849
                     /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 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 tacccttacg
     2821 acgtccccga ctacgcatat ccatatgatg ttccggatta cgcttgaact agttgaacgc
     2881 gtaccggtcc tagataagta atgatcataa tcagccatat cacatctgta gaggttttac
     2941 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3001 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3061 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3121 atgtatctta tcatgtctgg atcaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
     3181 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3241 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3301 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3361 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3421 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3481 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3541 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3601 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3661 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3721 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3781 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3841 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3901 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3961 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4021 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4081 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4141 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4201 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4261 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4321 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4381 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4441 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4501 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4561 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4621 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4681 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4741 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4801 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4861 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4921 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4981 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5041 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5101 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5161 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5221 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5281 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     5341 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     5401 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     5461 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     5521 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     5581 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     5641 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     5701 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     5761 aacgacggcc agtgccaagt tgggatcttt gcattggccc acggctctca ggatggggat
     5821 gctccccttc agcacccggt tccccttgga aactgatggt cctggctctg tggcatggca
     5881 gtggcactgt gaggagcccc ctaccagcag cacacagtgg ggttggcact gccacgctcc
     5941 ggatgccgcg ctctgatcca accccataat caagggaacc cgaattgccc catcattgcc
     6001 cccaccaccc ccatcctgcc gggccctcac accccacgct gccttctggt gacattcccc
     6061 agcccaaacc cacggcttca tggctaccgc ggggcatttc ccattgccgc cccattatca
     6121 gctctgcaca cctcccgctg tacccatgcc tcgtggctgc ccttctttga cgtataatct
     6181 tctaattaat acccggcctt gtcaaagtgg agcacaaacg ttaattaatt ccccagcagg
     6241 caggtaatta acagtgtgac tccctttttg ctgcgagtgg ggctgataca gagagatgtg
     6301 gcactatgga gcccacgggg tcctggcact gggtgcccac ggaggtcccc atgtgctgca
     6361 gtgtcaccgc ctccgaggag acagtattgt ccctgcggtg tccctgcagc tcagctctgt
     6421 ccacagggcc acctccagtt tggaggggac acaatgcagc cccgatgcaa cccatcctcg
     6481 cagcatccca gggacaaaga ccccactgca agaccgcaca cagggctggg tcccgctccc
     6541 ctaatatcta cagtgcttct tgcatggccc cttaatcaat gcagttaatc agcatgcgct
     6601 catgcaccgc tctggagctg caaagcccct cgcagcgctg ctcaccaaca ccgcgcaccg
     6661 ccccggccca gcctgcagca cgcgctgcaa acaggaaaga aacaaaatat tgcccaaatg
     6721 taggcaaagg cattcggctg ccttgacctc cgccgggccg ggccctgcct gactcagctc
     6781 cttactcagc gctcgcttcc tccctccggc tgccaccgcc gcagcgcaca ccctgacaaa
     6841 gagtggccct taacgggctc tgaggtgcac ccagcagtgc actcagcagt ccaagggccg
     6901 gcctggaggt ttgcaccgct acgtgctgac attagcattg aacttggccc tgggtagtgc
     6961 tgcaggccgg gcggggtggg tgtagagagt gcagcgcgcg ttgcacccgg tgccccttcc
     7021 cctcccttgc atcccagcag gctgcacccc agcaccaggc ccgtgcatgc atgctcctgg
     7081 tgttattgca gcctggtgca tgcatgcgtc ttgatggtgc agcgctgtgc atgcatcctc
     7141 cttggtgtgt agcagtctag tgcatgcata cccctcggtg ttattgctgc tctgtgcacg
     7201 cacgctcatt gtatcacttc atcccagtgc atgcactcac actggagcga ttgctgctcg
     7261 gtgcacgcac actcattgta tcactgcagc tcagtggctg cacgcacacc ggtgttattg
     7321 ctgctcggtg cgtgcatgca catcagtgtc gctgcagctc agtgcatgca cgctcattgc
     7381 ccatcgctat ccctgcctct cctgctggcg ctccccggga ggtgacttca aggggaccgc
     7441 aggaccacct cgggggtggg gggagggctg cacacgcgga ccccgctccc cctccccaac
     7501 aaagcactgt ggaatcaaaa aggggggagg ggggatggag gggcgcgtca cacccccgcc
     7561 ccacaccctc acctcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
     7621 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg
     7681 gggggggggg ggncgcgcgc caggcggggc ggggcggggc grggggcggg gcggggcgag
     7741 gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc
     7801 gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg g
//
