LOCUS       sequence-380538-        9289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Halo-tagged KAP 3B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9289)
  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 9289)
  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.Halo-KAP3B
FEATURES             Location/Qualifiers
     source          1..9289
                     /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             1324..2208
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     polyA_signal    4708..4842
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4745..4764)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4799..4818
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4873..4889)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4873..4889)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4886..4908)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4897..4913
                     /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(4921..4927,4928..4945,4946..4951))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4966..4987
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5104..5121)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5275..5863)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5355..5374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6034..6825,6826..6894))
                     /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     6657..6676
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6895..6999)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7067..7085
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7123..7145)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7245..7264
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7458..7480
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7472..7489
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7473..7489
                     /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 ggaccatggc tagcggagga
     1321 tccgaaatcg gtactggctt tccattcgac ccccattatg tggaagtcct gggcgagcgc
     1381 atgcactacg tcgatgttgg tccgcgcgat ggcacccctg tgctgttcct gcacggtaac
     1441 ccgacctcct cctacgtgtg gcgcaacatc atcccgcatg ttgcaccgac ccatcgctgc
     1501 attgctccag acctgatcgg tatgggcaaa tccgacaaac cagacctggg ttatttcttc
     1561 gacgaccacg tccgcttcat ggatgccttc atcgaagccc tgggtctgga agaggtcgtc
     1621 ctggtcattc acgactgggg ctccgctctg ggtttccact gggccaagcg caatccagag
     1681 cgcgtcaaag gtattgcatt tatggagttc atccgcccta tcccgacctg ggacgaatgg
     1741 ccagaatttg cccgcgagac cttccaggcc ttccgcacca ccgacgtcgg ccgcaagctg
     1801 atcatcgatc agaacgtttt tatcgagggt acgctgccga tgggtgtcgt ccgcccgctg
     1861 actgaagtcg agatggacca ttaccgcgag ccgttcctga atcctgttga ccgcgagcca
     1921 ctgtggcgct tcccaaacga gctgccaatc gccggtgagc cagcgaacat cgtcgcgctg
     1981 gtcgaagaat acatggactg gctgcaccag tcccctgtcc cgaagctgct gttctggggc
     2041 accccaggcg ttctgatccc accggccgaa gccgctcgcc tggccaaaag cctgcctaac
     2101 tgcaaggctg tggacatcgg cccgggtctg aatctgctgc aagaagacaa cccggacctg
     2161 atcggcagcg agatcgcgcg ctggctgtct actctggaga tttccgggct gtacggcgga
     2221 ggatccggag gcggctctgg cggagggctg tacaagatgc aaggggagga cgccagatac
     2281 ctcaaaagga aagttaaagg agggaatata gacgtgcatc catcagaaaa agcactcatt
     2341 gttcactatg aagtggaagc taccattctt ggagaaatgg gggaccccat gttgggagaa
     2401 cgaaaagaat gtcaaaaaat cattcgactt aagagtctca atgccaacac agatataact
     2461 tccctggcaa ggaaggtggt tgaagaatgt aaactcattc atccttcaaa actaaatgag
     2521 gtagaacagc tgttgtacta tctacagaac cgccgtgatt cattgtcagg aaaagagaaa
     2581 aaagaaaaat caagcaagcc taaagatcca cctccttttg aaggaatgga gattgatgaa
     2641 gttgctaaca ttaatgacat ggatgaatat attgagttat tatatgaaga tattcctgac
     2701 aaagttcggg gttctgcttt gatcctgcag cttgctcgaa atcctgataa cttggaagaa
     2761 ctactattga atgaaactgc ccttggtgca ttagcaaggg tcctgagaga agactggaag
     2821 caaagtgtcg agttagctac aaacataatt tacatctttt tttgtttctc cagcttttct
     2881 caatttcatg gacttattac tcactataaa attggagctc tgtgtatgaa tattattgat
     2941 catgagttaa aaagacatga gctttggcaa gaagaactct caaagaagaa gaaagctgtt
     3001 gatgaagacc ctgaaaacca aaccttgaga aaggattatg aaaaaacctt taaaaagtac
     3061 caggggcttg tggtaaaaca ggaacagcta ttacgagttg ctctttattt gcttctgaat
     3121 cttgctgagg atactcgtac cgaactgaaa atgaggaaca agaacatagt tcacatgttg
     3181 gtgaaagccc ttgatcggga caattttgag ctgctaattt tagttgtgtc attcttgaag
     3241 aaactcagca tttttatgga gaataaaaat gatatggtgg aaatggatat tgttgaaaaa
     3301 ctggtgaaaa tgataccttg tgagcatgaa gacctgctga atatcaccct ccgactttta
     3361 ctaaacctat cctttgacac aggactgagg aataagatgg tacaagttgg actgcttccc
     3421 aagctcactg cactcctagg caatgacaac tacaaacaaa tagcaatgtg tgttctttac
     3481 cacataagca tggatgaccg ctttaaatca atgtttgcat acactgactg tataccacag
     3541 ttaatgaaga tgctgtttga atgttcagat gaacgaattg acttggaact catttctttc
     3601 tgcattaatc ttgctgctaa caaaagaaac gttcagctta tctgtgaagg aaatgggctg
     3661 aagatgctca tgaagagggc tctgaagttt aaggatccat tgctgatgaa aatgattaga
     3721 aacatttctc agcatgatgg accaactaaa aatctgttta ttgattatgt tggggacctt
     3781 gcagcccaga tctctaatga tgaagaagag gagtttgtga ttgaatgttt gggaactctt
     3841 gcaaacttga ccattccaga cttagactgg gaattggttc ttaaagaata taagttggtt
     3901 ccatacctca aggataaact aaaaccaggt gctgcagaag atgatcttgt tttagaagtg
     3961 gttataatga ttggaactgt atccatggat gactcttgtg ctgcattgct agccaaatct
     4021 ggcataatcc ctgcactcat tgaattgcta aatgctcaac aagaagatga tgaatttgtg
     4081 tgtcagataa tttatgtctt ctaccagatg gttttccacc aagccacaag agacgtcata
     4141 atcaaggaaa cacaggctcc agcatatctc atagacctaa tgcatgataa gaataatgaa
     4201 atccgaaagg tctgtgataa tacattagat attatagcgg aatatgatga agaatgggct
     4261 aagaaaattc agagtgaaaa gtttcgctgg cataactctc agtggctgga gatggtagag
     4321 agtcgtcaga tggatgagag tgagcagtac ttgtatggtg atgatcgaat tgagccatac
     4381 attcatgaag gagatattct cgaaagacct gaccttttct acaactcaga tggattaatt
     4441 gcctctgaag gagccataag tcccgatttc ttcaatgatt accaccttca aaatggagat
     4501 gttgttgggc agcattcatt tcctggcagc acttttcatc caaggatttc aaagtgcttt
     4561 acaaatacgc actaacggac cgtctaatct agataagtaa tgatcataat cagccatatc
     4621 acatctgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     4681 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     4741 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4801 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tccccgggta ccgagctcga
     4861 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4921 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4981 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5041 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5101 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5161 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5221 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5281 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5341 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5401 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5461 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5521 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5581 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5641 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5701 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5761 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5821 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5881 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5941 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6001 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6061 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6121 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6181 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6241 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6301 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6361 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6421 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6481 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6541 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6601 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6661 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6721 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6781 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6841 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6901 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6961 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7021 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     7081 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     7141 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     7201 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     7261 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     7321 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     7381 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     7441 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagttggg
     7501 atctttgcat tggcccacgg ctctcaggat ggggatgctc cccttcagca cccggttccc
     7561 cttggaaact gatggtcctg gctctgtggc atggcagtgg cactgtgagg agccccctac
     7621 cagcagcaca cagtggggtt ggcactgcca cgctccggat gccgcgctct gatccaaccc
     7681 cataatcaag ggaacccgaa ttgccccatc attgccccca ccacccccat cctgccgggc
     7741 cctcacaccc cacgctgcct tctggtgaca ttccccagcc caaacccacg gcttcatggc
     7801 taccgcgggg catttcccat tgccgcccca ttatcagctc tgcacacctc ccgctgtacc
     7861 catgcctcgt ggctgccctt ctttgacgta taatcttcta attaataccc ggccttgtca
     7921 aagtggagca caaacgttaa ttaattcccc agcaggcagg taattaacag tgtgactccc
     7981 tttttgctgc gagtggggct gatacagaga gatgtggcac tatggagccc acggggtcct
     8041 ggcactgggt gcccacggag gtccccatgt gctgcagtgt caccgcctcc gaggagacag
     8101 tattgtccct gcggtgtccc tgcagctcag ctctgtccac agggccacct ccagtttgga
     8161 ggggacacaa tgcagccccg atgcaaccca tcctcgcagc atcccaggga caaagacccc
     8221 actgcaagac cgcacacagg gctgggtccc gctcccctaa tatctacagt gcttcttgca
     8281 tggcccctta atcaatgcag ttaatcagca tgcgctcatg caccgctctg gagctgcaaa
     8341 gcccctcgca gcgctgctca ccaacaccgc gcaccgcccc ggcccagcct gcagcacgcg
     8401 ctgcaaacag gaaagaaaca aaatattgcc caaatgtagg caaaggcatt cggctgcctt
     8461 gacctccgcc gggccgggcc ctgcctgact cagctcctta ctcagcgctc gcttcctccc
     8521 tccggctgcc accgccgcag cgcacaccct gacaaagagt ggcccttaac gggctctgag
     8581 gtgcacccag cagtgcactc agcagtccaa gggccggcct ggaggtttgc accgctacgt
     8641 gctgacatta gcattgaact tggccctggg tagtgctgca ggccgggcgg ggtgggtgta
     8701 gagagtgcag cgcgcgttgc acccggtgcc ccttcccctc ccttgcatcc cagcaggctg
     8761 caccccagca ccaggcccgt gcatgcatgc tcctggtgtt attgcagcct ggtgcatgca
     8821 tgcgtcttga tggtgcagcg ctgtgcatgc atcctccttg gtgtgtagca gtctagtgca
     8881 tgcatacccc tcggtgttat tgctgctctg tgcacgcacg ctcattgtat cacttcatcc
     8941 cagtgcatgc actcacactg gagcgattgc tgctcggtgc acgcacactc attgtatcac
     9001 tgcagctcag tggctgcacg cacaccggtg ttattgctgc tcggtgcgtg catgcacatc
     9061 agtgtcgctg cagctcagtg catgcacgct cattgcccat cgctatccct gcctctcctg
     9121 ctggcgctcc ccgggaggtg acttcaaggg gaccgcagga ccacctcggg ggtgggggga
     9181 gggctgcaca cgcggacccc gctccccctc cccaacaaag cactgtggaa tcaaaaaggg
     9241 gggagggggg atggaggggc gcgtcacacc cccgccccac accctcacc
//
