LOCUS       sequence-403673-        7566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Halo-tagged Kif 3B CC 391-592.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7566)
  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 7566)
  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 CC 391-592
FEATURES             Location/Qualifiers
     source          1..7566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        360..635
                     /label=chicken beta-actin promoter
     primer_bind     1578..1600
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             1683..2567
                     /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    3344..3478
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3381..3400)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3435..3454
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3509..3525)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3509..3525)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3522..3544)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3533..3549
                     /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(3557..3563,3564..3581,3582..3587))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3602..3623
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3740..3757)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3911..4499)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3991..4010)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4670..5461,5462..5530))
                     /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     5293..5312
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5531..5635)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5703..5721
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5759..5781)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5881..5900
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6094..6116
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6108..6125
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6109..6125
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cacttcatcc cagtgcatgc actcacactg gagcgattgc tgctcggtgc acgcacactc
       61 attgtatcac tgcagctcag tggctgcacg cacaccggtg ttattgctgc tcggtgcgtg
      121 catgcacatc agtgtcgctg cagctcagtg catgcacgct cattgcccat cgctatccct
      181 gcctctcctg ctggcgctcc ccgggaggtg acttcaaggg gaccgcagga ccacctcggg
      241 ggtgggggga gggctgcaca cgcggacccc gctccccctc cccaacaaag cactgtggaa
      301 tcaaaaaggg gggagggggg atggaggggc gcgtcacacc cccgccccac accctcacct
      361 cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat
      421 tttgtattta tttatttttt aattattttg tgcagcgatg ggggsggggg gggggggggc
      481 gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg
      541 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc
      601 ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc
      661 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
      721 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt
      781 aatgacggct cgtttctttt ctgtggctgc gtgaaagcct taaagggctc cgggagggcc
      841 ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc
      901 gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt
      961 gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg
     1021 ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt
     1081 gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc cctccccgag ttgctgagca
     1141 cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc gcggggctcg ccgtgccggg
     1201 cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggaggg
     1261 ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg
     1321 cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat
     1381 ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa
     1441 gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg
     1501 ccgtcccctt ctccatctcc agcctcgggg ctgtccgcag ggggacggct gccttcgggg
     1561 gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggggttt atatcttccc
     1621 ttctctgttc ctccgcagcc cccaagcttg gcgcgcctag gaccatggct agcggaggat
     1681 ccgaaatcgg tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca
     1741 tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc
     1801 cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca
     1861 ttgctccaga cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg
     1921 acgaccacgt ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc
     1981 tggtcattca cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc
     2041 gcgtcaaagg tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc
     2101 cagaatttgc ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga
     2161 tcatcgatca gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga
     2221 ctgaagtcga gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac
     2281 tgtggcgctt cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg
     2341 tcgaagaata catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca
     2401 ccccaggcgt tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact
     2461 gcaaggctgt ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga
     2521 tcggcagcga gatcgcgcgc tggctgtcta ctctggagat ttccgggctg tacggcggag
     2581 gatccggagg cggctctggc ggagggctgt acaagggggg tggggaagag gaggaggagg
     2641 agggagaaga gggtgaggag gacggggatg ataaggatga ttactggcgg gaacagcaag
     2701 aaaaactgga gattgagaag cgggccattg tagaggacca cagcttggtt gcagaggaga
     2761 agatgaggct gctgaaggag aaggagaaaa agatggagga cctgcggcga gagaaggatg
     2821 ctgcagagat gctgggtgcc aaaatcaagg ccatggagag taagctgctt gtcggaggaa
     2881 aaaatatagt agatcataca aacgaacagc agaagatcct ggagcaaaaa cgccaggaga
     2941 tcgcagagca gaaacgtcga gaaagagaaa tccagcagca aatggagagt cgagatgagg
     3001 agaccctgga actgaaagag acttacacct cactgcagca ggaggtagac atcaagacca
     3061 aaaagctcaa aaagctcttt tccaagcttc aggccgtgaa ggccgagatc cacgacctcc
     3121 aagaagaaca catcaaagag cgccaggagc tggagcagac gcagaatgaa ctcacccggg
     3181 agctaaagct caagcatctt attatagaaa actttattta aaatctagat aagtaatgat
     3241 cataatcagc catatcacat ctgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3301 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3361 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3421 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccc
     3481 cgggtaccga gctcgaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc
     3541 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3601 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3661 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3721 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3781 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3841 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3901 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3961 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4021 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4081 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4141 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4201 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4261 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4321 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4381 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4441 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4501 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4561 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4621 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4681 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4741 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4801 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4861 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4921 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4981 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5041 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5101 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5161 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5221 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5281 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5341 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5401 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5461 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5521 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5581 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5641 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     5701 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     5761 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     5821 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     5881 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     5941 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     6001 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     6061 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     6121 ccagtgccaa gttgggatct ttgcattggc ccacggctct caggatgggg atgctcccct
     6181 tcagcacccg gttccccttg gaaactgatg gtcctggctc tgtggcatgg cagtggcact
     6241 gtgaggagcc ccctaccagc agcacacagt ggggttggca ctgccacgct ccggatgccg
     6301 cgctctgatc caaccccata atcaagggaa cccgaattgc cccatcattg cccccaccac
     6361 ccccatcctg ccgggccctc acaccccacg ctgccttctg gtgacattcc ccagcccaaa
     6421 cccacggctt catggctacc gcggggcatt tcccattgcc gccccattat cagctctgca
     6481 cacctcccgc tgtacccatg cctcgtggct gcccttcttt gacgtataat cttctaatta
     6541 atacccggcc ttgtcaaagt ggagcacaaa cgttaattaa ttccccagca ggcaggtaat
     6601 taacagtgtg actccctttt tgctgcgagt ggggctgata cagagagatg tggcactatg
     6661 gagcccacgg ggtcctggca ctgggtgccc acggaggtcc ccatgtgctg cagtgtcacc
     6721 gcctccgagg agacagtatt gtccctgcgg tgtccctgca gctcagctct gtccacaggg
     6781 ccacctccag tttggagggg acacaatgca gccccgatgc aacccatcct cgcagcatcc
     6841 cagggacaaa gaccccactg caagaccgca cacagggctg ggtcccgctc ccctaatatc
     6901 tacagtgctt cttgcatggc cccttaatca atgcagttaa tcagcatgcg ctcatgcacc
     6961 gctctggagc tgcaaagccc ctcgcagcgc tgctcaccaa caccgcgcac cgccccggcc
     7021 cagcctgcag cacgcgctgc aaacaggaaa gaaacaaaat attgcccaaa tgtaggcaaa
     7081 ggcattcggc tgccttgacc tccgccgggc cgggccctgc ctgactcagc tccttactca
     7141 gcgctcgctt cctccctccg gctgccaccg ccgcagcgca caccctgaca aagagtggcc
     7201 cttaacgggc tctgaggtgc acccagcagt gcactcagca gtccaagggc cggcctggag
     7261 gtttgcaccg ctacgtgctg acattagcat tgaacttggc cctgggtagt gctgcaggcc
     7321 gggcggggtg ggtgtagaga gtgcagcgcg cgttgcaccc ggtgcccctt cccctccctt
     7381 gcatcccagc aggctgcacc ccagcaccag gcccgtgcat gcatgctcct ggtgttattg
     7441 cagcctggtg catgcatgcg tcttgatggt gcagcgctgt gcatgcatcc tccttggtgt
     7501 gtagcagtct agtgcatgca tacccctcgg tgttattgct gctctgtgca cgcacgctca
     7561 ttgtat
//
