LOCUS       sequence-374364-        8798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus expression of human KIF1C with R169W mutation and 6xHis 
            and GFP tags..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8798)
  AUTHORS   Siddiqui N, Roth D, Toleikis A, Zwetsloot AJ, Cross RA, Straube A
  TITLE     Force generation of KIF1C is impaired by pathogenic mutations.
  JOURNAL   Curr Biol. 2022 Sep 12;32(17):3862-3870.e6. doi: 
            10.1016/j.cub.2022.07.029. Epub 2022 Aug 11.
  PUBMED    35961316
REFERENCE   2  (bases 1 to 8798)
  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"; doi: 
            "10.1016/j.cub.2022.07.029"; journalName: "Curr Biol"; date: 
            "2022-09-12- 12"; volume: "32"; issue: "17"; pages: "3862-3870.e6"
COMMENT     Sequence Label: pFastBac-M13-6His-KIF1C_R169W-GFP
FEATURES             Location/Qualifiers
     source          1..8798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      226..235
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(232..234,235..237,238..951)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(232..234,235..237,238..948)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(277..298)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(538..557)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     885..906
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1074..1208
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1111..1130)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1165..1184
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1237..1402
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      1586..2040
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1673..1692)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1882..1903
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2066..2170
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2171..2239,2240..3031)
                     /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     complement(2389..2408)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3202..3790
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3691..3710
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3944..3961
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4095..4319
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4386..4919)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5089..5094,5095..5111,5112..5117))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        5469..5560
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5485..5502
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             5625..5642
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5664..5684
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     regulatory      7926..7935
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tccctggatg gagggggccg atcccgggga gcgggttctg cacagcctga accccagcac
       61 ttccagccca aaaagcacaa ctcttatccc cagccacccc aaccctaccc agcccagcgg
      121 cccccagggc cccgctaccc cccatacact actcccccac gaatgagacg gcagcgttct
      181 gcccctgacc tcaaggagag tggggcagct gtggatccac cggtcgccac catggtgagc
      241 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      301 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      361 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
      421 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
      481 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
      541 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
      601 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
      661 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
      721 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
      781 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
      841 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
      901 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccgc
      961 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     1021 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     1081 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1141 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     1201 tctggatctg atcactgata tcgcctagga gatccgaacc agataagtga aatctagttc
     1261 caaactattt tgtcattttt aattttcgta ttagcttacg acgctacacc cagttcccat
     1321 ctattttgtc actcttccct aaataatcct taaaaactcc atttccaccc ctcccagttc
     1381 ccaactattt tgtccgccca cagcggggca tttttcttcc tgttatgttt ttaatcaaac
     1441 atcctgccaa ctccatgtga caaaccgtca tcttcggcta ctttttctct gtcacagaat
     1501 gaaaattttt ctgtcatctc ttcgttatta atgtttgtaa ttgactgaat atcaacgctt
     1561 atttgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1621 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1681 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     1741 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     1801 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     1861 ttggagtcca cgttcttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     1921 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     1981 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     2041 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2101 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2161 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2221 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2281 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2341 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2401 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2461 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2521 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2581 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2641 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2701 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2761 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2821 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2881 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2941 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3001 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3061 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3121 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3181 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3241 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3301 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3361 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3421 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3481 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3541 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3601 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3661 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3721 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3781 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3841 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3901 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3961 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4021 ccgcatagac cagccgcgta acctggcaaa atcggttacg gttgagtaat aaatggatgc
     4081 cctgcgtaag cgggtgtggg cggacaataa agtcttaaac tgaacaaaat agatctaaac
     4141 tatgacaata aagtcttaaa ctagacagaa tagttgtaaa ctgaaatcag tccagttatg
     4201 ctgtgaaaaa gcatactgga cttttgttat ggctaaagca aactcttcat tttctgaagt
     4261 gcaaattgcc cgtcgtatta aagaggggcg tggccaaggg catggtaaag actatattcg
     4321 cggcgttgtg acaatttacc gaacaactcc gcggccggga agccgatctc ggcttgaacg
     4381 aattgttagg tggcggtact tgggtcgata tcaaagtgca tcacttcttc ccgtatgccc
     4441 aactttgtat agagagccac tgcgggatcg tcaccgtaat ctgcttgcac gtagatcaca
     4501 taagcaccaa gcgcgttggc ctcatgcttg aggagattga tgagcgcggt ggcaatgccc
     4561 tgcctccggt gctcgccgga gactgcgaga tcatagatat agatctcact acgcggctgc
     4621 tcaaacttgg gcagaacgta agccgcgaga gcgccaacaa ccgcttcttg gtcgaaggca
     4681 gcaagcgcga tgaatgtctt actacggagc aagttcccga ggtaatcgga gtccggctga
     4741 tgttgggagt aggtggctac gtctccgaac tcacgaccga aaagatcaag agcagcccgc
     4801 atggatttga cttggtcagg gccgagccta catgtgcgaa tgatgcccat acttgagcca
     4861 cctaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct gcgtaacatc
     4921 aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtaca
     4981 aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg
     5041 gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag tttacgaacc
     5101 gaacaggctt atgtcaactg ggttcgtgcc ttcatccgtt tccacggtgt gcgtcacccg
     5161 gcaaccttgg gcagcagcga agtcgaggca tttctgtcct ggctggcgaa cgagcgcaag
     5221 gtttcggtct ccacgcatcg tcaggcattg gcggccttgc tgttcttcta cggcaaggtg
     5281 ctgtgcacgg atctgccctg gcttcaggag atcggaagac ctcggccgtc gcggcgcttg
     5341 ccggtggtgc tgaccccgga tgaagtggtt cgcatcctcg gttttctgga aggcgagcat
     5401 cgtttgttcg cccaggactc tagctatagt tctagtggtt ggctacgtat actccggaat
     5461 attaatagat catggagata attaaaatga taaccatctc gcaaataaat aagtatttta
     5521 ctgttttcgt aacagttttg taataaaaaa acctataaat attccggatt attcataccc
     5581 ccaccatcgg gcgcggatct cggtccgaaa ccatgtcgta ctaccatcac catcaccatc
     5641 acgattacga tatcccaacg accgaaaacc tgtattttca gggcgccatg gcgcgcccgg
     5701 atctggctgg tgcctcggtg aaagtggcag tgagggttcg gccctttaac gcccgtgaga
     5761 ccagccagga tgccaagtgt gtggtcagca tgcagggcaa caccacctcc atcatcaatc
     5821 ctaaacagag caaggatgcc cccaaaagct tcacctttga ctactcctac tggtcacaca
     5881 cttcgacgga ggacccccag tttgcatctc agcagcaagt gtatcgggac atcggagaag
     5941 agatgctgct ccacgccttt gaaggctaca acgtgtgcat ctttgcctat gggcagaccg
     6001 gggctgggaa atcctatacc atgatggggc gacaggagcc agggcagcag ggcatcgtgc
     6061 cccagctctg tgaggacctc ttctctcgcg ttagtgagaa ccagagtgct cagctatcct
     6121 actctgtgga ggtgagctat atggagatct actgtgagcg ggtacgagac ctcttgaacc
     6181 ccaagagtcg gggttctctg cgggtctggg agcaccccat cctgggcccg tacgtgcagg
     6241 acctgtccaa attggctgtg acctcctacg cagacattgc tgacctcatg gactgtggaa
     6301 ataaagcacg gactgtggct gccaccaaca tgaatgagac cagcagccgt tcccatgccg
     6361 tctttaccat cgtcttcaca cagcgctgcc atgaccagct cacggggctg gactcggaga
     6421 aggtcagtaa gatcagtttg gtggaccttg ctgggagtga gcgagccgac tcctcagggg
     6481 cccggggcat gcgcctgaag gaaggagcca acatcaataa gtccctgact acactaggga
     6541 aagtgatctc ggcccttgca gatatgcaat caaagaagcg aaagtcggat tttatcccct
     6601 acagggactc tgtgctcacc tggctgctca aggaaaattt gggggggaac tcacgcacag
     6661 ccatgattgc agccctgagc cctgctgaca tcaattacga ggagactctc agcaccctca
     6721 ggtatgctga ccgcaccaag caaatccgct gcaatgccat catcaacgag gaccctaatg
     6781 cccggctgat tagagagctg caggaggaag tagcccggct gcgggaactg ctgatggctc
     6841 agggactgtc agcctctgct ctggaaggcc tgaagacgga agaagggagt gtcagaggcg
     6901 ccctgccagc tgtgtcatct cccccagctc cagtttcacc ctcatcaccc accacacata
     6961 atggggagct ggagccgtca ttctccccca acacggagtc ccagattggg cctgaggaag
     7021 ccatggagag gctgcaggag acagagaaga ttatagctga gctgaacgag acatgggagg
     7081 agaagctacg caagacagaa gccctgagga tggagagaga agcattgctg gctgagatgg
     7141 gggtggccgt ccgggaggat gggggaactg tgggcgtctt ctctccaaag aagactcccc
     7201 acctggtgaa cctgaacgaa gaccctctga tgtctgagtg tctgctctac cacatcaaag
     7261 atggcgtcac cagggtcggc caagtagata tggacatcaa gctgaccgga cagttcattc
     7321 gggagcaaca ctgtctgttc cggagcatcc cccagccaga tggagaagtg gtggtcactc
     7381 tggagccttg tgaaggagct gagacatatg tgaatgggaa gcttgtgacg gagccgctgg
     7441 tgctgaagtc agggaatagg attgtgatgg gcaagaacca cgttttccgc ttcaaccacc
     7501 cggagcaggc aaggctggaa cgggaacgag gggtcccccc acccccagga ccgccctctg
     7561 agccagtcga ctggaacttt gcccagaagg aactgctgga gcagcaaggc atcgacataa
     7621 agctggaaat ggagaagagg ctgcaggatc tggagaatca gtaccggaaa gaaaaggaag
     7681 aagccgatct tctgctggag cagcagcgac tgtatgcaga ctcggacagc ggggatgact
     7741 ctgacaagcg ctcttgtgaa gagagctgga ggctcatctc ctccttgcgg gagcagctgc
     7801 cgcccaccac ggtccagacc attgtcaaac gctgtggtct gcccagcagt ggcaagcgca
     7861 gggcccctcg cagggtttat cagatccccc agcgacgcag gctgcagggc aaagaccccc
     7921 gctgggccac catggctgac ctgaagatgc aggcggtgaa ggagatctgc tacgaggtgg
     7981 ccctggctga cttccgccac gggcgggctg agattgaggc cctggccgcc ctcaagatgc
     8041 gggagctgtg tcgcacctat ggcaagccag acggccccgg agacgcctgg agggctgtgg
     8101 cccgggatgt ctgggacact gtaggcgagg aggaaggagg tggagctggc agtggtggtg
     8161 gcagtgagga gggagcccga ggggcggagg tggaggacct ccgggcccac atcgacaagc
     8221 tgacggggat tctgcaggag gtgaagctgc agaacagcag caaggaccgg gagctgcagg
     8281 ccctgcggga ccgcatgctc cgcatggaga gggtcatccc cctggcccag gatcatgagg
     8341 atgagaatga agaaggtggt gaggtcccct gggccccgcc tgaaggatca gaggcagcag
     8401 aggaggcagc ccccagtgac cgcatgccgt cagcccggcc cccctcgcca ccactgtcaa
     8461 gctgggagcg ggtgtcacgg ctcatggagg aggaccctgc cttccgtcgt ggtcgtcttc
     8521 gctggctcaa gcaggagcag ctacggctgc agggactgca gggctctggg ggccggggcg
     8581 gggggctgcg caggccccca gcccgctttg tgccccctca cgactgcaag ctacgcttcc
     8641 ccttcaagag caacccccag caccgggagt cttggccagg gatggggagc ggggaggctc
     8701 caactccgct ccaaccccct gaggaggtca ctccccatcc agccacccct gcccgccggc
     8761 ctccgagtcc ccgaaggtcc caccatcccc gcaggaac
//
