LOCUS       sequence-380589-        8131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Halo-tagged Kif 3C tail 411-796.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8131)
  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 8131)
  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 tail 411-796
FEATURES             Location/Qualifiers
     source          1..8131
                     /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    3550..3684
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3587..3606)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3641..3660
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3715..3731)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3715..3731)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3728..3750)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3739..3755
                     /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(3763..3769,3770..3787,3788..3793))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3808..3829
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3946..3963)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4117..4705)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4197..4216)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4876..5667,5668..5736))
                     /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     5499..5518
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5737..5841)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5909..5927
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5965..5987)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6087..6106
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6300..6322
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6314..6331
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6315..6331
                     /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 tacaggagtc gtagaaagaa ggccgtgtct
     2281 gccccagccg gctatccaga ggggtccgtg attgaggcct gggtggccga agaggaagat
     2341 gataacaaca acaatcacca cccaccccag cccatcttgg aggcagcttt ggagaagaac
     2401 atggagaatt acctgcagga ccagaaggaa cggctggagg aggagaaggc tgccatccag
     2461 gatgaccgca gccttgtgag tgaggagaag cagaagcttc tagaagagaa ggagaagatg
     2521 ctggaggatc tacggcggga gcagcaggct acagagctgc ttgcagccaa gtacaaggcc
     2581 atggaaagca agctactcat tggggggcgg aacatcatgg atcacaccaa cgagcaacag
     2641 aagatgttgg aactgaagcg acaggagatt gcggaacaga aacgccgtga acgggaaatg
     2701 cagcaggaga tgctgctccg cgatgaagaa accatggagc tacgtggtac ctactcatcc
     2761 ctgcagcagg aggtggaagt taaaaccaag aaactcaaga agctctatgc caagctacag
     2821 gcggtgaagg ccgagatcca ggaccaacac gaagagtaca tccgcgtgcg gcaggacctg
     2881 gaggaggcgc agaatgagca gacccgggaa ctaaagctca agtacctcat cattgagaac
     2941 ttcatccccc ctgaggagaa gaacaagatc atgaaccgcc ttttcctgga ctgtgaggag
     3001 gagcagtgga ggttccagcc actggtgcca gctggagtga ataacagcca aatgaagaag
     3061 cgtccaactt ctgcggtggg ctacaagaga cctatcagcc agtatgctcg ggttgccatg
     3121 gcaatggggt cccatcccag atacagagct gaaaatataa tgtttttgga actggatgtg
     3181 tccccaccgg ctatctttga gatggagttt tctcatgacc aagagcaaga cccacgggta
     3241 ctacacatgg agaggctcat gaggttggac agctttctgg aaagaccgtc cacaactaaa
     3301 gtgcgaaagt cgagatcctg gtgccagagt cctcagcgga tgcctccacc ctccacagct
     3361 catgcatcca tgacctctgt tcctttgcac cctgcaacag tggtagacca tgactgacgg
     3421 accgtctaat ctagataagt aatgatcata atcagccata tcacatctgt agaggtttta
     3481 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     3541 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3601 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3661 aatgtatctt atcatgtctg gatccccggg taccgagctc gaattcgtaa tcatgtcata
     3721 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3781 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3841 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3901 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3961 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4021 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4081 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4141 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4201 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4261 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4321 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4381 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4441 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4501 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4561 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4621 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4681 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4741 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4801 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4861 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4921 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4981 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5041 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5101 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5161 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5221 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5281 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5341 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5401 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5461 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5521 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5581 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5641 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5701 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5761 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5821 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5881 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     5941 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     6001 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     6061 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     6121 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     6181 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     6241 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     6301 ccagtcacga cgttgtaaaa cgacggccag tgccaagttg ggatctttgc attggcccac
     6361 ggctctcagg atggggatgc tccccttcag cacccggttc cccttggaaa ctgatggtcc
     6421 tggctctgtg gcatggcagt ggcactgtga ggagccccct accagcagca cacagtgggg
     6481 ttggcactgc cacgctccgg atgccgcgct ctgatccaac cccataatca agggaacccg
     6541 aattgcccca tcattgcccc caccaccccc atcctgccgg gccctcacac cccacgctgc
     6601 cttctggtga cattccccag cccaaaccca cggcttcatg gctaccgcgg ggcatttccc
     6661 attgccgccc cattatcagc tctgcacacc tcccgctgta cccatgcctc gtggctgccc
     6721 ttctttgacg tataatcttc taattaatac ccggccttgt caaagtggag cacaaacgtt
     6781 aattaattcc ccagcaggca ggtaattaac agtgtgactc cctttttgct gcgagtgggg
     6841 ctgatacaga gagatgtggc actatggagc ccacggggtc ctggcactgg gtgcccacgg
     6901 aggtccccat gtgctgcagt gtcaccgcct ccgaggagac agtattgtcc ctgcggtgtc
     6961 cctgcagctc agctctgtcc acagggccac ctccagtttg gaggggacac aatgcagccc
     7021 cgatgcaacc catcctcgca gcatcccagg gacaaagacc ccactgcaag accgcacaca
     7081 gggctgggtc ccgctcccct aatatctaca gtgcttcttg catggcccct taatcaatgc
     7141 agttaatcag catgcgctca tgcaccgctc tggagctgca aagcccctcg cagcgctgct
     7201 caccaacacc gcgcaccgcc ccggcccagc ctgcagcacg cgctgcaaac aggaaagaaa
     7261 caaaatattg cccaaatgta ggcaaaggca ttcggctgcc ttgacctccg ccgggccggg
     7321 ccctgcctga ctcagctcct tactcagcgc tcgcttcctc cctccggctg ccaccgccgc
     7381 agcgcacacc ctgacaaaga gtggccctta acgggctctg aggtgcaccc agcagtgcac
     7441 tcagcagtcc aagggccggc ctggaggttt gcaccgctac gtgctgacat tagcattgaa
     7501 cttggccctg ggtagtgctg caggccgggc ggggtgggtg tagagagtgc agcgcgcgtt
     7561 gcacccggtg ccccttcccc tcccttgcat cccagcaggc tgcaccccag caccaggccc
     7621 gtgcatgcat gctcctggtg ttattgcagc ctggtgcatg catgcgtctt gatggtgcag
     7681 cgctgtgcat gcatcctcct tggtgtgtag cagtctagtg catgcatacc cctcggtgtt
     7741 attgctgctc tgtgcacgca cgctcattgt atcacttcat cccagtgcat gcactcacac
     7801 tggagcgatt gctgctcggt gcacgcacac tcattgtatc actgcagctc agtggctgca
     7861 cgcacaccgg tgttattgct gctcggtgcg tgcatgcaca tcagtgtcgc tgcagctcag
     7921 tgcatgcacg ctcattgccc atcgctatcc ctgcctctcc tgctggcgct ccccgggagg
     7981 tgacttcaag gggaccgcag gaccacctcg ggggtggggg gagggctgca cacgcggacc
     8041 ccgctccccc tccccaacaa agcactgtgg aatcaaaaag gggggagggg ggatggaggg
     8101 gcgcgtcaca cccccgcccc acaccctcac c
//
