LOCUS       sequence-414191-        7825 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Halo-tagged Kif 3A tail 419-702.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7825)
  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 7825)
  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-KIF3A tail 419-702
FEATURES             Location/Qualifiers
     source          1..7825
                     /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    3244..3378
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3281..3300)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3335..3354
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3409..3425)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3409..3425)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3422..3444)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3433..3449
                     /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(3457..3463,3464..3481,3482..3487))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3502..3523
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3640..3657)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3811..4399)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3891..3910)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4570..5361,5362..5430))
                     /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     5193..5212
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5431..5535)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5603..5621
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5659..5681)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5781..5800
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5994..6016
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6008..6025
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6009..6025
                     /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 tacaagccag acaagatgat tgaaatgcaa
     2281 gcaaaaattg atgaggagag aaaagcactt gaaacaaagc tcgacatgga agaagaagaa
     2341 agaaacaagg ctagagctga attagagaaa cgggaaaaag atcttcttaa agcccaacaa
     2401 gagcatcagt ctttgctgga aaaattatct gccctggaaa agaaggtaat tgttggtggg
     2461 gttgacttgt tggccaaagc tgaggaacaa gagaaacttc ttgaagaatc taacatggaa
     2521 ctggaagaaa ggaggaaaag ggcagagcaa cttcgcagag aacttgagga aaaagagcaa
     2581 gaacgcttgg atattgaaga aaaatatacc agtttgcaag aggaagcaca gggaaagacc
     2641 aagaagttaa agaaagtttg gactatgctg atggctgcaa agtcagagat ggctgatctc
     2701 caacaagaac atcagaggga aattgaaggc ctactggaga acattcggca acttagccgg
     2761 gagcttcgac ttcagatgct tattattgat aactttatac ctcgggatta tcaggaaatg
     2821 attgaaaact atgtccattg gaatgaagac ataggagaat ggcagctaaa atgtgttgct
     2881 tatacaggaa ataacatgag gaagcaaacc ccagtacctg ataaaaagga gaaagatccc
     2941 tttgaggtgg acctttctca cgtgtatctt gcctatactg aggagagtct gcgtcagtct
     3001 ttgatgaaac tagaaagacc acgaacttca aaggggaaag caaggccaaa gacagggaga
     3061 agaaagcgtt ctgcaaagcc tgaaactgta attgactctc tgctgcagta acggaccgtc
     3121 taatctagat aagtaatgat cataatcagc catatcacat ctgtagaggt tttacttgct
     3181 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     3241 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3301 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3361 tcttatcatg tctggatccc cgggtaccga gctcgaattc gtaatcatgt catagctgtt
     3421 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3481 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3541 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3601 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3661 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3721 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3781 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3841 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3901 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3961 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4021 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4081 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4141 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4201 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4261 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4321 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4381 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4441 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4501 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4561 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4621 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4681 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4741 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4801 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4861 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4921 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4981 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5041 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5101 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5161 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5221 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5281 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5341 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5401 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5461 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5521 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     5581 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     5641 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     5701 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     5761 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     5821 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     5881 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     5941 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     6001 acgacgttgt aaaacgacgg ccagtgccaa gttgggatct ttgcattggc ccacggctct
     6061 caggatgggg atgctcccct tcagcacccg gttccccttg gaaactgatg gtcctggctc
     6121 tgtggcatgg cagtggcact gtgaggagcc ccctaccagc agcacacagt ggggttggca
     6181 ctgccacgct ccggatgccg cgctctgatc caaccccata atcaagggaa cccgaattgc
     6241 cccatcattg cccccaccac ccccatcctg ccgggccctc acaccccacg ctgccttctg
     6301 gtgacattcc ccagcccaaa cccacggctt catggctacc gcggggcatt tcccattgcc
     6361 gccccattat cagctctgca cacctcccgc tgtacccatg cctcgtggct gcccttcttt
     6421 gacgtataat cttctaatta atacccggcc ttgtcaaagt ggagcacaaa cgttaattaa
     6481 ttccccagca ggcaggtaat taacagtgtg actccctttt tgctgcgagt ggggctgata
     6541 cagagagatg tggcactatg gagcccacgg ggtcctggca ctgggtgccc acggaggtcc
     6601 ccatgtgctg cagtgtcacc gcctccgagg agacagtatt gtccctgcgg tgtccctgca
     6661 gctcagctct gtccacaggg ccacctccag tttggagggg acacaatgca gccccgatgc
     6721 aacccatcct cgcagcatcc cagggacaaa gaccccactg caagaccgca cacagggctg
     6781 ggtcccgctc ccctaatatc tacagtgctt cttgcatggc cccttaatca atgcagttaa
     6841 tcagcatgcg ctcatgcacc gctctggagc tgcaaagccc ctcgcagcgc tgctcaccaa
     6901 caccgcgcac cgccccggcc cagcctgcag cacgcgctgc aaacaggaaa gaaacaaaat
     6961 attgcccaaa tgtaggcaaa ggcattcggc tgccttgacc tccgccgggc cgggccctgc
     7021 ctgactcagc tccttactca gcgctcgctt cctccctccg gctgccaccg ccgcagcgca
     7081 caccctgaca aagagtggcc cttaacgggc tctgaggtgc acccagcagt gcactcagca
     7141 gtccaagggc cggcctggag gtttgcaccg ctacgtgctg acattagcat tgaacttggc
     7201 cctgggtagt gctgcaggcc gggcggggtg ggtgtagaga gtgcagcgcg cgttgcaccc
     7261 ggtgcccctt cccctccctt gcatcccagc aggctgcacc ccagcaccag gcccgtgcat
     7321 gcatgctcct ggtgttattg cagcctggtg catgcatgcg tcttgatggt gcagcgctgt
     7381 gcatgcatcc tccttggtgt gtagcagtct agtgcatgca tacccctcgg tgttattgct
     7441 gctctgtgca cgcacgctca ttgtatcact tcatcccagt gcatgcactc acactggagc
     7501 gattgctgct cggtgcacgc acactcattg tatcactgca gctcagtggc tgcacgcaca
     7561 ccggtgttat tgctgctcgg tgcgtgcatg cacatcagtg tcgctgcagc tcagtgcatg
     7621 cacgctcatt gcccatcgct atccctgcct ctcctgctgg cgctccccgg gaggtgactt
     7681 caaggggacc gcaggaccac ctcgggggtg gggggagggc tgcacacgcg gaccccgctc
     7741 cccctcccca acaaagcact gtggaatcaa aaagggggga ggggggatgg aggggcgcgt
     7801 cacacccccg ccccacaccc tcacc
//
