LOCUS       sequence-380261-        7315 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Halo-tagged Kif 3A DD 590-702.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7315)
  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 7315)
  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 DD 590-702
FEATURES             Location/Qualifiers
     source          1..7315
                     /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    2734..2868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2771..2790)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2825..2844
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2899..2915)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2899..2915)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2912..2934)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2923..2939
                     /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(2947..2953,2954..2971,2972..2977))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2992..3013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3130..3147)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3301..3889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3381..3400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4060..4851,4852..4920))
                     /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     4683..4702
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4921..5025)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5093..5111
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5149..5171)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5271..5290
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5484..5506
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5498..5515
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5499..5515
                     /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 tacatccgac ttcagatgct tattattgat
     2281 aactttatac ctcgggatta tcaggaaatg attgaaaact atgtccattg gaatgaagac
     2341 ataggagaat ggcagctaaa atgtgttgct tatacaggaa ataacatgag gaagcaaacc
     2401 ccagtacctg ataaaaagga gaaagatccc tttgaggtgg acctttctca cgtgtatctt
     2461 gcctatactg aggagagtct gcgtcagtct ttgatgaaac tagaaagacc acgaacttca
     2521 aaggggaaag caaggccaaa gacagggaga agaaagcgtt ctgcaaagcc tgaaactgta
     2581 attgactctc tgctgcagta acggaccgtc taatctagat aagtaatgat cataatcagc
     2641 catatcacat ctgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     2701 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2761 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2821 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccc cgggtaccga
     2881 gctcgaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2941 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3001 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3061 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3121 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3181 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3241 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3301 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3361 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3421 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3481 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3541 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3601 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3661 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3721 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3781 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3841 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3901 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3961 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4021 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4081 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4141 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4201 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4261 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4321 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4381 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4441 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4501 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4561 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4621 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4681 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4741 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4801 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4861 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4921 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4981 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5041 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     5101 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     5161 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     5221 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     5281 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     5341 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5401 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     5461 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
     5521 gttgggatct ttgcattggc ccacggctct caggatgggg atgctcccct tcagcacccg
     5581 gttccccttg gaaactgatg gtcctggctc tgtggcatgg cagtggcact gtgaggagcc
     5641 ccctaccagc agcacacagt ggggttggca ctgccacgct ccggatgccg cgctctgatc
     5701 caaccccata atcaagggaa cccgaattgc cccatcattg cccccaccac ccccatcctg
     5761 ccgggccctc acaccccacg ctgccttctg gtgacattcc ccagcccaaa cccacggctt
     5821 catggctacc gcggggcatt tcccattgcc gccccattat cagctctgca cacctcccgc
     5881 tgtacccatg cctcgtggct gcccttcttt gacgtataat cttctaatta atacccggcc
     5941 ttgtcaaagt ggagcacaaa cgttaattaa ttccccagca ggcaggtaat taacagtgtg
     6001 actccctttt tgctgcgagt ggggctgata cagagagatg tggcactatg gagcccacgg
     6061 ggtcctggca ctgggtgccc acggaggtcc ccatgtgctg cagtgtcacc gcctccgagg
     6121 agacagtatt gtccctgcgg tgtccctgca gctcagctct gtccacaggg ccacctccag
     6181 tttggagggg acacaatgca gccccgatgc aacccatcct cgcagcatcc cagggacaaa
     6241 gaccccactg caagaccgca cacagggctg ggtcccgctc ccctaatatc tacagtgctt
     6301 cttgcatggc cccttaatca atgcagttaa tcagcatgcg ctcatgcacc gctctggagc
     6361 tgcaaagccc ctcgcagcgc tgctcaccaa caccgcgcac cgccccggcc cagcctgcag
     6421 cacgcgctgc aaacaggaaa gaaacaaaat attgcccaaa tgtaggcaaa ggcattcggc
     6481 tgccttgacc tccgccgggc cgggccctgc ctgactcagc tccttactca gcgctcgctt
     6541 cctccctccg gctgccaccg ccgcagcgca caccctgaca aagagtggcc cttaacgggc
     6601 tctgaggtgc acccagcagt gcactcagca gtccaagggc cggcctggag gtttgcaccg
     6661 ctacgtgctg acattagcat tgaacttggc cctgggtagt gctgcaggcc gggcggggtg
     6721 ggtgtagaga gtgcagcgcg cgttgcaccc ggtgcccctt cccctccctt gcatcccagc
     6781 aggctgcacc ccagcaccag gcccgtgcat gcatgctcct ggtgttattg cagcctggtg
     6841 catgcatgcg tcttgatggt gcagcgctgt gcatgcatcc tccttggtgt gtagcagtct
     6901 agtgcatgca tacccctcgg tgttattgct gctctgtgca cgcacgctca ttgtatcact
     6961 tcatcccagt gcatgcactc acactggagc gattgctgct cggtgcacgc acactcattg
     7021 tatcactgca gctcagtggc tgcacgcaca ccggtgttat tgctgctcgg tgcgtgcatg
     7081 cacatcagtg tcgctgcagc tcagtgcatg cacgctcatt gcccatcgct atccctgcct
     7141 ctcctgctgg cgctccccgg gaggtgactt caaggggacc gcaggaccac ctcgggggtg
     7201 gggggagggc tgcacacgcg gaccccgctc cccctcccca acaaagcact gtggaatcaa
     7261 aaagggggga ggggggatgg aggggcgcgt cacacccccg ccccacaccc tcacc
//
