LOCUS       sequence-380569-        7519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Halo-tagged Kif 3C DD 616-796.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7519)
  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 7519)
  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 DD 616-796
FEATURES             Location/Qualifiers
     source          1..7519
                     /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    2938..3072
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2975..2994)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3029..3048
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3103..3119)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3103..3119)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3116..3138)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3127..3143
                     /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(3151..3157,3158..3175,3176..3181))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3196..3217
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3334..3351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3505..4093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3585..3604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4264..5055,5056..5124))
                     /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     4887..4906
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5125..5229)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5297..5315
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5353..5375)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5475..5494
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5688..5710
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5702..5719
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5703..5719
                     /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 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 tacactcggc aggacctgga ggaggcgcag
     2281 aatgagcaga cccgggaact aaagctcaag tacctcatca ttgagaactt catcccccct
     2341 gaggagaaga acaagatcat gaaccgcctt ttcctggact gtgaggagga gcagtggagg
     2401 ttccagccac tggtgccagc tggagtgaat aacagccaaa tgaagaagcg tccaacttct
     2461 gcggtgggct acaagagacc tatcagccag tatgctcggg ttgccatggc aatggggtcc
     2521 catcccagat acagagctga aaatataatg tttttggaac tggatgtgtc cccaccggct
     2581 atctttgaga tggagttttc tcatgaccaa gagcaagacc cacgggtact acacatggag
     2641 aggctcatga ggttggacag ctttctggaa agaccgtcca caactaaagt gcgaaagtcg
     2701 agatcctggt gccagagtcc tcagcggatg cctccaccct ccacagctca tgcatccatg
     2761 acctctgttc ctttgcaccc tgcaacagtg gtagaccatg actgacggac cgtctaatct
     2821 agataagtaa tgatcataat cagccatatc acatctgtag aggttttact tgctttaaaa
     2881 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2941 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3001 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     3061 catgtctgga tccccgggta ccgagctcga attcgtaatc atgtcatagc tgtttcctgt
     3121 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3181 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3241 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3301 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3361 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3421 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3481 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3541 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3601 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3661 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3721 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3781 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3841 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3901 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3961 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4021 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4081 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4141 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4201 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4261 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4321 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4381 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4441 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4501 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4561 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4621 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4681 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4741 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4801 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4861 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4921 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4981 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5041 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5101 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5161 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5221 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     5281 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     5341 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     5401 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     5461 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     5521 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     5581 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     5641 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     5701 ttgtaaaacg acggccagtg ccaagttggg atctttgcat tggcccacgg ctctcaggat
     5761 ggggatgctc cccttcagca cccggttccc cttggaaact gatggtcctg gctctgtggc
     5821 atggcagtgg cactgtgagg agccccctac cagcagcaca cagtggggtt ggcactgcca
     5881 cgctccggat gccgcgctct gatccaaccc cataatcaag ggaacccgaa ttgccccatc
     5941 attgccccca ccacccccat cctgccgggc cctcacaccc cacgctgcct tctggtgaca
     6001 ttccccagcc caaacccacg gcttcatggc taccgcgggg catttcccat tgccgcccca
     6061 ttatcagctc tgcacacctc ccgctgtacc catgcctcgt ggctgccctt ctttgacgta
     6121 taatcttcta attaataccc ggccttgtca aagtggagca caaacgttaa ttaattcccc
     6181 agcaggcagg taattaacag tgtgactccc tttttgctgc gagtggggct gatacagaga
     6241 gatgtggcac tatggagccc acggggtcct ggcactgggt gcccacggag gtccccatgt
     6301 gctgcagtgt caccgcctcc gaggagacag tattgtccct gcggtgtccc tgcagctcag
     6361 ctctgtccac agggccacct ccagtttgga ggggacacaa tgcagccccg atgcaaccca
     6421 tcctcgcagc atcccaggga caaagacccc actgcaagac cgcacacagg gctgggtccc
     6481 gctcccctaa tatctacagt gcttcttgca tggcccctta atcaatgcag ttaatcagca
     6541 tgcgctcatg caccgctctg gagctgcaaa gcccctcgca gcgctgctca ccaacaccgc
     6601 gcaccgcccc ggcccagcct gcagcacgcg ctgcaaacag gaaagaaaca aaatattgcc
     6661 caaatgtagg caaaggcatt cggctgcctt gacctccgcc gggccgggcc ctgcctgact
     6721 cagctcctta ctcagcgctc gcttcctccc tccggctgcc accgccgcag cgcacaccct
     6781 gacaaagagt ggcccttaac gggctctgag gtgcacccag cagtgcactc agcagtccaa
     6841 gggccggcct ggaggtttgc accgctacgt gctgacatta gcattgaact tggccctggg
     6901 tagtgctgca ggccgggcgg ggtgggtgta gagagtgcag cgcgcgttgc acccggtgcc
     6961 ccttcccctc ccttgcatcc cagcaggctg caccccagca ccaggcccgt gcatgcatgc
     7021 tcctggtgtt attgcagcct ggtgcatgca tgcgtcttga tggtgcagcg ctgtgcatgc
     7081 atcctccttg gtgtgtagca gtctagtgca tgcatacccc tcggtgttat tgctgctctg
     7141 tgcacgcacg ctcattgtat cacttcatcc cagtgcatgc actcacactg gagcgattgc
     7201 tgctcggtgc acgcacactc attgtatcac tgcagctcag tggctgcacg cacaccggtg
     7261 ttattgctgc tcggtgcgtg catgcacatc agtgtcgctg cagctcagtg catgcacgct
     7321 cattgcccat cgctatccct gcctctcctg ctggcgctcc ccgggaggtg acttcaaggg
     7381 gaccgcagga ccacctcggg ggtgggggga gggctgcaca cgcggacccc gctccccctc
     7441 cccaacaaag cactgtggaa tcaaaaaggg gggagggggg atggaggggc gcgtcacacc
     7501 cccgccccac accctcacc
//
