LOCUS       sequence-380567-        8788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of KAP 3B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8788)
  AUTHORS   Garbouchian A, Montgomery A, Gilbert SP, Bentley M
  TITLE     KAP is the neuronal organelle adaptor for Kinesin-2 KIF3AB and 
            KIF3AC.
  JOURNAL   Mol Biol Cell. 2022 Oct 6:mbcE22080336. doi: 
            10.1091/mbc.E22-08-0336.
  PUBMED    36200888
REFERENCE   2  (bases 1 to 8788)
  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: "Mol Biol 
            Cell. 2022 Oct 6:mbcE22080336. doi: 10.1091/mbc.E22-08-0336."
COMMENT     Sequence Label: pBa.FRB-3myc-KAP3B
FEATURES             Location/Qualifiers
     source          1..8788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..276
                     /label=chicken beta-actin promoter
     primer_bind     1219..1241
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             1321..1599
                     /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"
     CDS             1606..1632
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4207..4341
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4244..4263)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4298..4317
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4372..4388)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4372..4388)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4385..4407)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4396..4412
                     /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(4420..4426,4427..4444,4445..4450))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4465..4486
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4603..4620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4774..5362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4854..4873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5533..6324,6325..6393))
                     /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     6156..6175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6394..6498)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6566..6584
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6622..6644)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6744..6763
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6957..6979
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6971..6988
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6972..6988
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
       61 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg
      121 cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg
      181 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
      241 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
      301 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      361 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
      421 taatgacggc tcgtttcttt tctgtggctg cgtgaaagcc ttaaagggct ccgggagggc
      481 cctttgtgcg ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc
      541 cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg
      601 tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg
      661 gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg
      721 tgtgggcgcg gcggtcgggc tgtaaccccc ccctgcaccc ccctccccga gttgctgagc
      781 acggcccggc ttcgggtgcg gggctccgtg cggggcgtgg cgcggggctc gccgtgccgg
      841 gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg
      901 gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc
      961 gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa
     1021 tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga
     1081 agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc
     1141 gccgtcccct tctccatctc cagcctcggg gctgtccgca gggggacggc tgccttcggg
     1201 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggggtt tatatcttcc
     1261 cttctctgtt cctccgcagc ccccaagctt ggcgcgccta ggcctaccat ggcttccaga
     1321 atcctctggc atgagatgtg gcatgaaggc ctggaagagg catctcgttt gtactttggg
     1381 gaaaggaacg tgaaaggcat gtttgaggtg ctggagccct tgcatgctat gatggaacgg
     1441 ggcccccaga ctctgaagga aacatccttt aatcaggcct atggtcgaga tttaatggag
     1501 gcccaagagt ggtgcaggaa gtacatgaaa tcagggaatg tcaaggacct cctccaagcc
     1561 tgggacctct attatcatgt gttccgacga atctcaaaga ctagttatcc gtacgacgta
     1621 ccagactacg cacagaagct gatctcagag gaggacctgc aaaaacttat ttctgaagag
     1681 gatctgcaaa aacttatttc tgaagaagat ctgggcggag gatccggagg cggctctggc
     1741 ggagggctgt acaagatgca aggggaggac gccagatacc tcaaaaggaa agttaaagga
     1801 gggaatatag acgtgcatcc atcagaaaaa gcactcattg ttcactatga agtggaagct
     1861 accattcttg gagaaatggg ggaccccatg ttgggagaac gaaaagaatg tcaaaaaatc
     1921 attcgactta agagtctcaa tgccaacaca gatataactt ccctggcaag gaaggtggtt
     1981 gaagaatgta aactcattca tccttcaaaa ctaaatgagg tagaacagct gttgtactat
     2041 ctacagaacc gccgtgattc attgtcagga aaagagaaaa aagaaaaatc aagcaagcct
     2101 aaagatccac ctccttttga aggaatggag attgatgaag ttgctaacat taatgacatg
     2161 gatgaatata ttgagttatt atatgaagat attcctgaca aagttcgggg ttctgctttg
     2221 atcctgcagc ttgctcgaaa tcctgataac ttggaagaac tactattgaa tgaaactgcc
     2281 cttggtgcat tagcaagggt cctgagagaa gactggaagc aaagtgtcga gttagctaca
     2341 aacataattt acatcttttt ttgtttctcc agcttttctc aatttcatgg acttattact
     2401 cactataaaa ttggagctct gtgtatgaat attattgatc atgagttaaa aagacatgag
     2461 ctttggcaag aagaactctc aaagaagaag aaagctgttg atgaagaccc tgaaaaccaa
     2521 accttgagaa aggattatga aaaaaccttt aaaaagtacc aggggcttgt ggtaaaacag
     2581 gaacagctat tacgagttgc tctttatttg cttctgaatc ttgctgagga tactcgtacc
     2641 gaactgaaaa tgaggaacaa gaacatagtt cacatgttgg tgaaagccct tgatcgggac
     2701 aattttgagc tgctaatttt agttgtgtca ttcttgaaga aactcagcat ttttatggag
     2761 aataaaaatg atatggtgga aatggatatt gttgaaaaac tggtgaaaat gataccttgt
     2821 gagcatgaag acctgctgaa tatcaccctc cgacttttac taaacctatc ctttgacaca
     2881 ggactgagga ataagatggt acaagttgga ctgcttccca agctcactgc actcctaggc
     2941 aatgacaact acaaacaaat agcaatgtgt gttctttacc acataagcat ggatgaccgc
     3001 tttaaatcaa tgtttgcata cactgactgt ataccacagt taatgaagat gctgtttgaa
     3061 tgttcagatg aacgaattga cttggaactc atttctttct gcattaatct tgctgctaac
     3121 aaaagaaacg ttcagcttat ctgtgaagga aatgggctga agatgctcat gaagagggct
     3181 ctgaagttta aggatccatt gctgatgaaa atgattagaa acatttctca gcatgatgga
     3241 ccaactaaaa atctgtttat tgattatgtt ggggaccttg cagcccagat ctctaatgat
     3301 gaagaagagg agtttgtgat tgaatgtttg ggaactcttg caaacttgac cattccagac
     3361 ttagactggg aattggttct taaagaatat aagttggttc catacctcaa ggataaacta
     3421 aaaccaggtg ctgcagaaga tgatcttgtt ttagaagtgg ttataatgat tggaactgta
     3481 tccatggatg actcttgtgc tgcattgcta gccaaatctg gcataatccc tgcactcatt
     3541 gaattgctaa atgctcaaca agaagatgat gaatttgtgt gtcagataat ttatgtcttc
     3601 taccagatgg ttttccacca agccacaaga gacgtcataa tcaaggaaac acaggctcca
     3661 gcatatctca tagacctaat gcatgataag aataatgaaa tccgaaaggt ctgtgataat
     3721 acattagata ttatagcgga atatgatgaa gaatgggcta agaaaattca gagtgaaaag
     3781 tttcgctggc ataactctca gtggctggag atggtagaga gtcgtcagat ggatgagagt
     3841 gagcagtact tgtatggtga tgatcgaatt gagccataca ttcatgaagg agatattctc
     3901 gaaagacctg accttttcta caactcagat ggattaattg cctctgaagg agccataagt
     3961 cccgatttct tcaatgatta ccaccttcaa aatggagatg ttgttgggca gcattcattt
     4021 cctggcagca cttttcatcc aaggatttca aagtgcttta caaatacgca ctaacggacc
     4081 gtctaatcta gataagtaat gatcataatc agccatatca catctgtaga ggttttactt
     4141 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     4201 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4261 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4321 gtatcttatc atgtctggat ccccgggtac cgagctcgaa ttcgtaatca tgtcatagct
     4381 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4441 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4501 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4561 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4621 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4681 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4741 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4801 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4861 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4921 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4981 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5041 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5101 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5161 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5221 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5281 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5341 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5401 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5461 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5521 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5581 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5641 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5701 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5761 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5821 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5881 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5941 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6001 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6061 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6121 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6181 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6241 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6301 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6361 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6421 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6481 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     6541 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     6601 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     6661 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     6721 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     6781 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     6841 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     6901 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     6961 gtcacgacgt tgtaaaacga cggccagtgc caagttggga tctttgcatt ggcccacggc
     7021 tctcaggatg gggatgctcc ccttcagcac ccggttcccc ttggaaactg atggtcctgg
     7081 ctctgtggca tggcagtggc actgtgagga gccccctacc agcagcacac agtggggttg
     7141 gcactgccac gctccggatg ccgcgctctg atccaacccc ataatcaagg gaacccgaat
     7201 tgccccatca ttgcccccac cacccccatc ctgccgggcc ctcacacccc acgctgcctt
     7261 ctggtgacat tccccagccc aaacccacgg cttcatggct accgcggggc atttcccatt
     7321 gccgccccat tatcagctct gcacacctcc cgctgtaccc atgcctcgtg gctgcccttc
     7381 tttgacgtat aatcttctaa ttaatacccg gccttgtcaa agtggagcac aaacgttaat
     7441 taattcccca gcaggcaggt aattaacagt gtgactccct ttttgctgcg agtggggctg
     7501 atacagagag atgtggcact atggagccca cggggtcctg gcactgggtg cccacggagg
     7561 tccccatgtg ctgcagtgtc accgcctccg aggagacagt attgtccctg cggtgtccct
     7621 gcagctcagc tctgtccaca gggccacctc cagtttggag gggacacaat gcagccccga
     7681 tgcaacccat cctcgcagca tcccagggac aaagacccca ctgcaagacc gcacacaggg
     7741 ctgggtcccg ctcccctaat atctacagtg cttcttgcat ggccccttaa tcaatgcagt
     7801 taatcagcat gcgctcatgc accgctctgg agctgcaaag cccctcgcag cgctgctcac
     7861 caacaccgcg caccgccccg gcccagcctg cagcacgcgc tgcaaacagg aaagaaacaa
     7921 aatattgccc aaatgtaggc aaaggcattc ggctgccttg acctccgccg ggccgggccc
     7981 tgcctgactc agctccttac tcagcgctcg cttcctccct ccggctgcca ccgccgcagc
     8041 gcacaccctg acaaagagtg gcccttaacg ggctctgagg tgcacccagc agtgcactca
     8101 gcagtccaag ggccggcctg gaggtttgca ccgctacgtg ctgacattag cattgaactt
     8161 ggccctgggt agtgctgcag gccgggcggg gtgggtgtag agagtgcagc gcgcgttgca
     8221 cccggtgccc cttcccctcc cttgcatccc agcaggctgc accccagcac caggcccgtg
     8281 catgcatgct cctggtgtta ttgcagcctg gtgcatgcat gcgtcttgat ggtgcagcgc
     8341 tgtgcatgca tcctccttgg tgtgtagcag tctagtgcat gcatacccct cggtgttatt
     8401 gctgctctgt gcacgcacgc tcattgtatc acttcatccc agtgcatgca ctcacactgg
     8461 agcgattgct gctcggtgca cgcacactca ttgtatcact gcagctcagt ggctgcacgc
     8521 acaccggtgt tattgctgct cggtgcgtgc atgcacatca gtgtcgctgc agctcagtgc
     8581 atgcacgctc attgcccatc gctatccctg cctctcctgc tggcgctccc cgggaggtga
     8641 cttcaagggg accgcaggac cacctcgggg gtggggggag ggctgcacac gcggaccccg
     8701 ctccccctcc ccaacaaagc actgtggaat caaaaagggg ggagggggga tggaggggcg
     8761 cgtcacaccc ccgccccaca ccctcacc
//
