LOCUS       sequence-403665-        7476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Halo-tagged Kif 3A CC 419-589.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7476)
  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 7476)
  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 CC 419-589
FEATURES             Location/Qualifiers
     source          1..7476
                     /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    2895..3029
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2932..2951)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2986..3005
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3060..3076)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3060..3076)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3073..3095)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3084..3100
                     /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(3108..3114,3115..3132,3133..3138))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3153..3174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3291..3308)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3462..4050)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3542..3561)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4221..5012,5013..5081))
                     /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     4844..4863
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5082..5186)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5254..5272
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5310..5332)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5432..5451
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5645..5667
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5659..5676
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5660..5676
                     /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 gggggsgggg 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 gagctttaat ttaatctaga taagtaatga tcataatcag ccatatcaca tctgtagagg
     2821 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     2881 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2941 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3001 tcatcaatgt atcttatcat gtctggatcc ccgggtaccg agctcgaatt cgtaatcatg
     3061 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3121 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3181 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3241 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3301 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3361 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3421 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3481 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3541 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3601 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3661 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3721 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3781 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3841 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3901 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3961 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4021 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4081 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4141 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4201 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4261 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4321 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4381 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4441 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4501 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4561 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4621 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4681 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4741 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4801 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4861 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4921 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4981 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5041 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5101 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5161 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     5221 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     5281 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     5341 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     5401 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     5461 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     5521 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     5581 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     5641 ttttcccagt cacgacgttg taaaacgacg gccagtgcca agttgggatc tttgcattgg
     5701 cccacggctc tcaggatggg gatgctcccc ttcagcaccc ggttcccctt ggaaactgat
     5761 ggtcctggct ctgtggcatg gcagtggcac tgtgaggagc cccctaccag cagcacacag
     5821 tggggttggc actgccacgc tccggatgcc gcgctctgat ccaaccccat aatcaaggga
     5881 acccgaattg ccccatcatt gcccccacca cccccatcct gccgggccct cacaccccac
     5941 gctgccttct ggtgacattc cccagcccaa acccacggct tcatggctac cgcggggcat
     6001 ttcccattgc cgccccatta tcagctctgc acacctcccg ctgtacccat gcctcgtggc
     6061 tgcccttctt tgacgtataa tcttctaatt aatacccggc cttgtcaaag tggagcacaa
     6121 acgttaatta attccccagc aggcaggtaa ttaacagtgt gactcccttt ttgctgcgag
     6181 tggggctgat acagagagat gtggcactat ggagcccacg gggtcctggc actgggtgcc
     6241 cacggaggtc cccatgtgct gcagtgtcac cgcctccgag gagacagtat tgtccctgcg
     6301 gtgtccctgc agctcagctc tgtccacagg gccacctcca gtttggaggg gacacaatgc
     6361 agccccgatg caacccatcc tcgcagcatc ccagggacaa agaccccact gcaagaccgc
     6421 acacagggct gggtcccgct cccctaatat ctacagtgct tcttgcatgg ccccttaatc
     6481 aatgcagtta atcagcatgc gctcatgcac cgctctggag ctgcaaagcc cctcgcagcg
     6541 ctgctcacca acaccgcgca ccgccccggc ccagcctgca gcacgcgctg caaacaggaa
     6601 agaaacaaaa tattgcccaa atgtaggcaa aggcattcgg ctgccttgac ctccgccggg
     6661 ccgggccctg cctgactcag ctccttactc agcgctcgct tcctccctcc ggctgccacc
     6721 gccgcagcgc acaccctgac aaagagtggc ccttaacggg ctctgaggtg cacccagcag
     6781 tgcactcagc agtccaaggg ccggcctgga ggtttgcacc gctacgtgct gacattagca
     6841 ttgaacttgg ccctgggtag tgctgcaggc cgggcggggt gggtgtagag agtgcagcgc
     6901 gcgttgcacc cggtgcccct tcccctccct tgcatcccag caggctgcac cccagcacca
     6961 ggcccgtgca tgcatgctcc tggtgttatt gcagcctggt gcatgcatgc gtcttgatgg
     7021 tgcagcgctg tgcatgcatc ctccttggtg tgtagcagtc tagtgcatgc atacccctcg
     7081 gtgttattgc tgctctgtgc acgcacgctc attgtatcac ttcatcccag tgcatgcact
     7141 cacactggag cgattgctgc tcggtgcacg cacactcatt gtatcactgc agctcagtgg
     7201 ctgcacgcac accggtgtta ttgctgctcg gtgcgtgcat gcacatcagt gtcgctgcag
     7261 ctcagtgcat gcacgctcat tgcccatcgc tatccctgcc tctcctgctg gcgctccccg
     7321 ggaggtgact tcaaggggac cgcaggacca cctcgggggt ggggggaggg ctgcacacgc
     7381 ggaccccgct ccccctcccc aacaaagcac tgtggaatca aaaagggggg aggggggatg
     7441 gaggggcgcg tcacaccccc gccccacacc ctcacc
//
