LOCUS       sequence-407044-        7574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Halo-tagged Kif 3C CC 411-615.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7574)
  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 7574)
  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-KIF3C CC 411-615
FEATURES             Location/Qualifiers
     source          1..7574
                     /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    3352..3486
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3389..3408)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3443..3462
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3517..3533)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3517..3533)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3530..3552)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3541..3557
                     /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(3565..3571,3572..3589,3590..3595))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3610..3631
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3748..3765)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3919..4507)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3999..4018)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4678..5469,5470..5538))
                     /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     5301..5320
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5539..5643)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5711..5729
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5767..5789)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5889..5908
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6102..6124
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6116..6133
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6117..6133
                     /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 ggggcggggg 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 acagtcgtag aaagaaggcc gtgtctgccc
     2641 cagccggcta tccagagggg tccgtgattg aggcctgggt ggccgaagag gaagatgata
     2701 acaacaacaa tcaccaccca ccccagccca tcttggaggc agctttggag aagaacatgg
     2761 agaattacct gcaggaccag aaggaacggc tggaggagga gaaggctgcc atccaggatg
     2821 accgcagcct tgtgagtgag gagaagcaga agcttctaga agagaaggag aagatgctgg
     2881 aggatctacg gcgggagcag caggctacag agctgcttgc agccaagtac aaggccatgg
     2941 aaagcaagct actcattggg gggcggaaca tcatggatca caccaacgag caacagaaga
     3001 tgttggaact gaagcgacag gagattgcgg aacagaaacg ccgtgaacgg gaaatgcagc
     3061 aggagatgct gctccgcgat gaagaaacca tggagctacg tggtacctac tcatccctgc
     3121 agcaggaggt ggaagttaaa accaagaaac tcaagaagct ctatgccaag ctacaggcgg
     3181 tgaaggccga gatccaggac caacacgaag agtacatccg cgtgcgtcta atctagataa
     3241 gtaatgatca taatcagcca tatcacatct gtagaggttt tacttgcttt aaaaaacctc
     3301 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     3361 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3421 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3481 tggatccccg ggtaccgagc tcgaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
     3541 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3601 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3661 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3721 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3781 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3841 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3901 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3961 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4021 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4081 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4141 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4201 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4261 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4321 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4381 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4441 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4501 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4561 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4621 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4681 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4741 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4801 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4861 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4921 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4981 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5041 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5101 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5161 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5221 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5281 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5341 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5401 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5461 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5521 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5581 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5641 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     5701 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     5761 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     5821 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     5881 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     5941 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     6001 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     6061 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     6121 aacgacggcc agtgccaagt tgggatcttt gcattggccc acggctctca ggatggggat
     6181 gctccccttc agcacccggt tccccttgga aactgatggt cctggctctg tggcatggca
     6241 gtggcactgt gaggagcccc ctaccagcag cacacagtgg ggttggcact gccacgctcc
     6301 ggatgccgcg ctctgatcca accccataat caagggaacc cgaattgccc catcattgcc
     6361 cccaccaccc ccatcctgcc gggccctcac accccacgct gccttctggt gacattcccc
     6421 agcccaaacc cacggcttca tggctaccgc ggggcatttc ccattgccgc cccattatca
     6481 gctctgcaca cctcccgctg tacccatgcc tcgtggctgc ccttctttga cgtataatct
     6541 tctaattaat acccggcctt gtcaaagtgg agcacaaacg ttaattaatt ccccagcagg
     6601 caggtaatta acagtgtgac tccctttttg ctgcgagtgg ggctgataca gagagatgtg
     6661 gcactatgga gcccacgggg tcctggcact gggtgcccac ggaggtcccc atgtgctgca
     6721 gtgtcaccgc ctccgaggag acagtattgt ccctgcggtg tccctgcagc tcagctctgt
     6781 ccacagggcc acctccagtt tggaggggac acaatgcagc cccgatgcaa cccatcctcg
     6841 cagcatccca gggacaaaga ccccactgca agaccgcaca cagggctggg tcccgctccc
     6901 ctaatatcta cagtgcttct tgcatggccc cttaatcaat gcagttaatc agcatgcgct
     6961 catgcaccgc tctggagctg caaagcccct cgcagcgctg ctcaccaaca ccgcgcaccg
     7021 ccccggccca gcctgcagca cgcgctgcaa acaggaaaga aacaaaatat tgcccaaatg
     7081 taggcaaagg cattcggctg ccttgacctc cgccgggccg ggccctgcct gactcagctc
     7141 cttactcagc gctcgcttcc tccctccggc tgccaccgcc gcagcgcaca ccctgacaaa
     7201 gagtggccct taacgggctc tgaggtgcac ccagcagtgc actcagcagt ccaagggccg
     7261 gcctggaggt ttgcaccgct acgtgctgac attagcattg aacttggccc tgggtagtgc
     7321 tgcaggccgg gcggggtggg tgtagagagt gcagcgcgcg ttgcacccgg tgccccttcc
     7381 cctcccttgc atcccagcag gctgcacccc agcaccaggc ccgtgcatgc atgctcctgg
     7441 tgttattgca gcctggtgca tgcatgcgtc ttgatggtgc agcgctgtgc atgcatcctc
     7501 cttggtgtgt agcagtctag tgcatgcata cccctcggtg ttattgctgc tctgtgcacg
     7561 cacgctcatt gtat
//
