LOCUS       sequence-380419-        7430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Halo-tagged Kif 3B DD 593-747.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7430)
  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 7430)
  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 DD 593-747
FEATURES             Location/Qualifiers
     source          1..7430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..277
                     /label=chicken beta-actin promoter
     primer_bind     1220..1242
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     CDS             1325..2209
                     /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    2849..2983
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2886..2905)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2940..2959
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3014..3030)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3014..3030)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3027..3049)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3038..3054
                     /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(3062..3068,3069..3086,3087..3092))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3107..3128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3245..3262)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3416..4004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3496..3515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4175..4966,4967..5035))
                     /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     4798..4817
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5036..5140)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5208..5226
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5264..5286)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5386..5405
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5599..5621
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5613..5630
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5614..5630
                     /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 gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc
      181 ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg
      241 gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg
      301 ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt
      361 actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt
      421 ttaatgacgg ctcgtttctt ttctgtggct gcgtgaaagc cttaaagggc tccgggaggg
      481 ccctttgtgc gggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg
      541 ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt
      601 gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg
      661 ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg
      721 gtgtgggcgc ggcggtcggg ctgtaacccc cccctgcacc cccctccccg agttgctgag
      781 cacggcccgg cttcgggtgc ggggctccgt gcggggcgtg gcgcggggct cgccgtgccg
      841 ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag
      901 ggctcggggg aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc
      961 cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa
     1021 atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg
     1081 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
     1141 cgccgtcccc ttctccatct ccagcctcgg ggctgtccgc agggggacgg ctgccttcgg
     1201 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggggt ttatatcttc
     1261 ccttctctgt tcctccgcag cccccaagct tggcgcgcct aggaccatgg ctagcggagg
     1321 atccgaaatc ggtactggct ttccattcga cccccattat gtggaagtcc tgggcgagcg
     1381 catgcactac gtcgatgttg gtccgcgcga tggcacccct gtgctgttcc tgcacggtaa
     1441 cccgacctcc tcctacgtgt ggcgcaacat catcccgcat gttgcaccga cccatcgctg
     1501 cattgctcca gacctgatcg gtatgggcaa atccgacaaa ccagacctgg gttatttctt
     1561 cgacgaccac gtccgcttca tggatgcctt catcgaagcc ctgggtctgg aagaggtcgt
     1621 cctggtcatt cacgactggg gctccgctct gggtttccac tgggccaagc gcaatccaga
     1681 gcgcgtcaaa ggtattgcat ttatggagtt catccgccct atcccgacct gggacgaatg
     1741 gccagaattt gcccgcgaga ccttccaggc cttccgcacc accgacgtcg gccgcaagct
     1801 gatcatcgat cagaacgttt ttatcgaggg tacgctgccg atgggtgtcg tccgcccgct
     1861 gactgaagtc gagatggacc attaccgcga gccgttcctg aatcctgttg accgcgagcc
     1921 actgtggcgc ttcccaaacg agctgccaat cgccggtgag ccagcgaaca tcgtcgcgct
     1981 ggtcgaagaa tacatggact ggctgcacca gtcccctgtc ccgaagctgc tgttctgggg
     2041 caccccaggc gttctgatcc caccggccga agccgctcgc ctggccaaaa gcctgcctaa
     2101 ctgcaaggct gtggacatcg gcccgggtct gaatctgctg caagaagaca acccggacct
     2161 gatcggcagc gagatcgcgc gctggctgtc tactctggag atttccgggc tgtacggcgg
     2221 aggatccgga ggcggctctg gcggagggct gtacaaaccc ttggaagaga agaataaaat
     2281 tatgaataga tccttttttg atgatgaaga agaccattgg aaattacatc ctataaccag
     2341 actggaaaac cagcagatga tgaagcggcc tgtctctgct gtgggataca agagacctct
     2401 gagccagcat gcacggatgt ccatgatgat tcggccagag ccccggtaca gggcggagaa
     2461 catcatgctc ttggagttag atatgcccag ccggacaacc agagactacg agggcccagc
     2521 catctctccc aaggttcagg ctgcgctgga tgcagctctg caggatgaag atgagataca
     2581 ggtggatgcc tcatcctttg aaagtactgc cagcagaaaa cccaaggcca ggcccaagag
     2641 tggcaggaaa tcaggatcct cctcttcctc ctcaggaaac cctgcatctc agttttaccc
     2701 acagtctcgg gggctggttc ccaagtaatc tagataagta atgatcataa tcagccatat
     2761 cacatctgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     2821 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     2881 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2941 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccccgggt accgagctcg
     3001 aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3061 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3121 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3181 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3241 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3301 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3361 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3421 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3481 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3541 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3601 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3661 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3721 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3781 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3841 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3901 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3961 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4021 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4081 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4141 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4201 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4261 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4321 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4381 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4441 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4501 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4561 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4621 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4681 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4741 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4801 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4861 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4921 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4981 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5041 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5101 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5161 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     5221 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     5281 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     5341 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     5401 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     5461 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     5521 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     5581 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagttgg
     5641 gatctttgca ttggcccacg gctctcagga tggggatgct ccccttcagc acccggttcc
     5701 ccttggaaac tgatggtcct ggctctgtgg catggcagtg gcactgtgag gagcccccta
     5761 ccagcagcac acagtggggt tggcactgcc acgctccgga tgccgcgctc tgatccaacc
     5821 ccataatcaa gggaacccga attgccccat cattgccccc accaccccca tcctgccggg
     5881 ccctcacacc ccacgctgcc ttctggtgac attccccagc ccaaacccac ggcttcatgg
     5941 ctaccgcggg gcatttccca ttgccgcccc attatcagct ctgcacacct cccgctgtac
     6001 ccatgcctcg tggctgccct tctttgacgt ataatcttct aattaatacc cggccttgtc
     6061 aaagtggagc acaaacgtta attaattccc cagcaggcag gtaattaaca gtgtgactcc
     6121 ctttttgctg cgagtggggc tgatacagag agatgtggca ctatggagcc cacggggtcc
     6181 tggcactggg tgcccacgga ggtccccatg tgctgcagtg tcaccgcctc cgaggagaca
     6241 gtattgtccc tgcggtgtcc ctgcagctca gctctgtcca cagggccacc tccagtttgg
     6301 aggggacaca atgcagcccc gatgcaaccc atcctcgcag catcccaggg acaaagaccc
     6361 cactgcaaga ccgcacacag ggctgggtcc cgctccccta atatctacag tgcttcttgc
     6421 atggcccctt aatcaatgca gttaatcagc atgcgctcat gcaccgctct ggagctgcaa
     6481 agcccctcgc agcgctgctc accaacaccg cgcaccgccc cggcccagcc tgcagcacgc
     6541 gctgcaaaca ggaaagaaac aaaatattgc ccaaatgtag gcaaaggcat tcggctgcct
     6601 tgacctccgc cgggccgggc cctgcctgac tcagctcctt actcagcgct cgcttcctcc
     6661 ctccggctgc caccgccgca gcgcacaccc tgacaaagag tggcccttaa cgggctctga
     6721 ggtgcaccca gcagtgcact cagcagtcca agggccggcc tggaggtttg caccgctacg
     6781 tgctgacatt agcattgaac ttggccctgg gtagtgctgc aggccgggcg gggtgggtgt
     6841 agagagtgca gcgcgcgttg cacccggtgc cccttcccct cccttgcatc ccagcaggct
     6901 gcaccccagc accaggcccg tgcatgcatg ctcctggtgt tattgcagcc tggtgcatgc
     6961 atgcgtcttg atggtgcagc gctgtgcatg catcctcctt ggtgtgtagc agtctagtgc
     7021 atgcataccc ctcggtgtta ttgctgctct gtgcacgcac gctcattgta tcacttcatc
     7081 ccagtgcatg cactcacact ggagcgattg ctgctcggtg cacgcacact cattgtatca
     7141 ctgcagctca gtggctgcac gcacaccggt gttattgctg ctcggtgcgt gcatgcacat
     7201 cagtgtcgct gcagctcagt gcatgcacgc tcattgccca tcgctatccc tgcctctcct
     7261 gctggcgctc cccgggaggt gacttcaagg ggaccgcagg accacctcgg gggtgggggg
     7321 agggctgcac acgcggaccc cgctccccct ccccaacaaa gcactgtgga atcaaaaagg
     7381 ggggaggggg gatggagggg cgcgtcacac ccccgcccca caccctcacc
//
