LOCUS       sequence-380185-        8032 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Halo-tagged Kif 3B tail 391-747.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8032)
  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 8032)
  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-KIF3B tail 391-747
FEATURES             Location/Qualifiers
     source          1..8032
                     /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    3451..3585
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3488..3507)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3542..3561
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3616..3632)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3616..3632)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3629..3651)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3640..3656
                     /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(3664..3670,3671..3688,3689..3694))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3709..3730
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3847..3864)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4018..4606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4098..4117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4777..5568,5569..5637))
                     /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     5400..5419
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5638..5742)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5810..5828
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5866..5888)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5988..6007
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6201..6223
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6215..6232
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6216..6232
                     /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 gcggggcgrg 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 tacaagggag gtggggaaga ggaggaggag
     2281 gagggagaag agggtgagga ggacggggat gataaggatg attactggcg ggaacagcaa
     2341 gaaaaactgg agattgagaa gcgggccatt gtagaggacc acagcttggt tgcagaggag
     2401 aagatgaggc tgctgaagga gaaggagaaa aagatggagg acctgcggcg agagaaggat
     2461 gctgcagaga tgctgggtgc caaaatcaag gccatggaga gtaagctgct tgtcggagga
     2521 aaaaatatag tagatcatac aaacgaacag cagaagatcc tggagcaaaa acgccaggag
     2581 atcgcagagc agaaacgtcg agaaagagaa atccagcagc aaatggagag tcgagatgag
     2641 gagaccctgg aactgaaaga gacttacacc tcactgcagc aggaggtaga catcaagacc
     2701 aaaaagctca aaaagctctt ttccaagctt caggccgtga aggccgagat ccacgacctc
     2761 caagaagaac acatcaaaga gcgccaggag ctggagcaga cgcagaatga actcacccgg
     2821 gagctaaagc tcaagcatct tattatagaa aactttattc ccttggaaga gaagaataaa
     2881 attatgaata gatccttttt tgatgatgaa gaagaccatt ggaaattaca tcctataacc
     2941 agactggaaa accagcagat gatgaagcgg cctgtctctg ctgtgggata caagagacct
     3001 ctgagccagc atgcacggat gtccatgatg attcggccag agccccggta cagggcggag
     3061 aacatcatgc tcttggagtt agatatgccc agccggacaa ccagagacta cgagggccca
     3121 gccatctctc ccaaggttca ggctgcgctg gatgcagctc tgcaggatga agatgagata
     3181 caggtggatg cctcatcctt tgaaagtact gccagcagaa aacccaaggc caggcccaag
     3241 agtggcagga aatcaggatc ctcctcttcc tcctcaggaa accctgcatc tcagttttac
     3301 ccacagtctc gggggctggt tcccaagtaa tctagataag taatgatcat aatcagccat
     3361 atcacatctg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     3421 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3481 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3541 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccccgg gtaccgagct
     3601 cgaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3661 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3721 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3781 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3841 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3901 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3961 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4021 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4081 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4141 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4201 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4261 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4321 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4381 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4441 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4501 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4561 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4621 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4681 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4741 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4801 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4861 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4921 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4981 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5041 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5101 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5161 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5221 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5281 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5341 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5401 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5461 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5521 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5581 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5641 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5701 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5761 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     5821 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     5881 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     5941 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     6001 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     6061 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     6121 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     6181 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagtt
     6241 gggatctttg cattggccca cggctctcag gatggggatg ctccccttca gcacccggtt
     6301 ccccttggaa actgatggtc ctggctctgt ggcatggcag tggcactgtg aggagccccc
     6361 taccagcagc acacagtggg gttggcactg ccacgctccg gatgccgcgc tctgatccaa
     6421 ccccataatc aagggaaccc gaattgcccc atcattgccc ccaccacccc catcctgccg
     6481 ggccctcaca ccccacgctg ccttctggtg acattcccca gcccaaaccc acggcttcat
     6541 ggctaccgcg gggcatttcc cattgccgcc ccattatcag ctctgcacac ctcccgctgt
     6601 acccatgcct cgtggctgcc cttctttgac gtataatctt ctaattaata cccggccttg
     6661 tcaaagtgga gcacaaacgt taattaattc cccagcaggc aggtaattaa cagtgtgact
     6721 ccctttttgc tgcgagtggg gctgatacag agagatgtgg cactatggag cccacggggt
     6781 cctggcactg ggtgcccacg gaggtcccca tgtgctgcag tgtcaccgcc tccgaggaga
     6841 cagtattgtc cctgcggtgt ccctgcagct cagctctgtc cacagggcca cctccagttt
     6901 ggaggggaca caatgcagcc ccgatgcaac ccatcctcgc agcatcccag ggacaaagac
     6961 cccactgcaa gaccgcacac agggctgggt cccgctcccc taatatctac agtgcttctt
     7021 gcatggcccc ttaatcaatg cagttaatca gcatgcgctc atgcaccgct ctggagctgc
     7081 aaagcccctc gcagcgctgc tcaccaacac cgcgcaccgc cccggcccag cctgcagcac
     7141 gcgctgcaaa caggaaagaa acaaaatatt gcccaaatgt aggcaaaggc attcggctgc
     7201 cttgacctcc gccgggccgg gccctgcctg actcagctcc ttactcagcg ctcgcttcct
     7261 ccctccggct gccaccgccg cagcgcacac cctgacaaag agtggccctt aacgggctct
     7321 gaggtgcacc cagcagtgca ctcagcagtc caagggccgg cctggaggtt tgcaccgcta
     7381 cgtgctgaca ttagcattga acttggccct gggtagtgct gcaggccggg cggggtgggt
     7441 gtagagagtg cagcgcgcgt tgcacccggt gccccttccc ctcccttgca tcccagcagg
     7501 ctgcacccca gcaccaggcc cgtgcatgca tgctcctggt gttattgcag cctggtgcat
     7561 gcatgcgtct tgatggtgca gcgctgtgca tgcatcctcc ttggtgtgta gcagtctagt
     7621 gcatgcatac ccctcggtgt tattgctgct ctgtgcacgc acgctcattg tatcacttca
     7681 tcccagtgca tgcactcaca ctggagcgat tgctgctcgg tgcacgcaca ctcattgtat
     7741 cactgcagct cagtggctgc acgcacaccg gtgttattgc tgctcggtgc gtgcatgcac
     7801 atcagtgtcg ctgcagctca gtgcatgcac gctcattgcc catcgctatc cctgcctctc
     7861 ctgctggcgc tccccgggag gtgacttcaa ggggaccgca ggaccacctc gggggtgggg
     7921 ggagggctgc acacgcggac cccgctcccc ctccccaaca aagcactgtg gaatcaaaaa
     7981 ggggggaggg gggatggagg ggcgcgtcac acccccgccc cacaccctca cc
//
