LOCUS       sequence-449873-        8699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Drosophila UNC-104(G618R) in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8699)
  AUTHORS   Niwa S, Watanabe T, Chiba K
  TITLE     The FHA domain is essential for the autoinhibition of KIF1A/UNC-104.
  JOURNAL   J Cell Sci. 2024 Sep 6:jcs.262017. doi: 10.1242/jcs.262017.
  PUBMED    39239883
REFERENCE   2  (bases 1 to 8699)
  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: "J Cell Sci.
            2024 Sep 6:jcs.262017. doi: 10.1242/jcs.262017."
COMMENT     Sequence Label: pKC28
FEATURES             Location/Qualifiers
     source          1..8699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        67..158
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     83..100
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             join(5254..5256,5257..5967)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             join(5980..6003,6004..6039,6040..6063)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="WSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     polyA_signal    6212..6346
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6249..6268)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6303..6322
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6467..6632
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      6822..7410
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7311..7330
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  7545..7768
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(7835..8368)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(8557..8562,8563..8579,8580..8585))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(8656..8689)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 aacggtccta aggtagcgag tttaaacact agtatcgatt cgcgacctac tccggaatat
       61 taatagatca tggagataat taaaatgata accatctcgc aaataaataa gtattttact
      121 gttttcgtaa cagttttgta ataaaaaaac ctataaatat tccggattat tcataccgtc
      181 ccaccatcgg gcgcggatcc cggtccgaag cgcgcggaat tcatgtcgtc ggttaaggtg
      241 gcggtgcgag tgcgcccctt caactcgcgc gaaatagcca gggagtcgaa atgcatcatc
      301 gagatggcag gagccacaac ggccatcacc aatccaaagg tgccgcccaa cacaagcgat
      361 tcagtaaagc gcttcaactt tgattactcc tactggtcac atgatcacca cgatgccgat
      421 ttctctacac aatcgatggt gtacaaggac attggcgagg agatgttaca gcactccttc
      481 gatggctaca atgtctgtat ctttgcctat ggccagactg gagctggaaa gtcgtatacc
      541 atgatgggca ggcaggagga gcagcaagag ggcataatac caatgatttg caaggatctg
      601 ttcactcgca tacaggatac tgagaccgat gacctcaagt actcggttga ggtttcctat
      661 atggaaatct attgcgagcg agtcagggat ttgttgaatc cgaaaaacaa gggcaatctg
      721 cgtgtgaggg agcatcccct tttgggtccc tatgtggagg atttgtccaa actagccgtt
      781 accgattacc aggacattca cgatctcatt gatgaaggca acaaagctcg aactgtggca
      841 gctactaaca tgaacgaaac cagttcccgc tctcatgccg tctttacaat cttctttacc
      901 caacgtcgtc atgatctgat gacaaatttg accacagaaa aggtatccaa gataagcttg
      961 gtagacttgg ccgggtcgga acgagcagat tccactggtg ccaagggcac tcgcttgaag
     1021 gagggagcca acattaacaa gtccttgacc acattgggca aagttatctc agctttggcg
     1081 gaagtgtctg cttccaagaa aaagaacacc aagaaggcag attttattcc gtaccgtgat
     1141 tcggccttga cctggctgtt acgtgaaaac ttgggaggaa attcgaagac agctatgatt
     1201 gcggctatct caccagcaga tattaactac gatgaaaccc tcagcacact gcgctatgcg
     1261 gatcgtgcca agcaaattgt ttgcaaggct gttgtcaatg aagatgccaa tgccaagctg
     1321 attcgagaac tcaaggagga gatccaaaag ctgcgggact tactcaaagc cgagggcatt
     1381 gaagtgcagg aagaggatga gctcaccaag tcaacggtga tcaagtcgcc cactaagtca
     1441 cgtaatcgta atggatccac cacggagatg gctgtggatc agctgcaggc cagcgagaaa
     1501 cttattgcag agctcaacga gacctgggag gagaaactta agcgcaccga ggagattcgc
     1561 gtgcagcgag aggcggtctt cgccgaaatg ggtgtggctg tcaaggaaga tggaataaca
     1621 gttggcgtat tctcaccgaa gaagactccg catttggtca acctaaacga ggatcccaat
     1681 ctgtctgagt gtctgcttta ctacatcaag gagggtctaa ctcggttggg tacccatgaa
     1741 gcaaatgtgc cccaggacat tcagctctcc ggatcgcaca tcctcaagga gcactgcacc
     1801 tttgagaaca agaacagcac ggttacattg ctgccacaca aggatgctat catctatgta
     1861 aatggacgca agttggttga accggaggtt cttaagaccg gttctcgcgt gatcctcgga
     1921 aagaaccacg tgttccgctt taccaatcca gaacaggcac gcgaattacg ggataagatc
     1981 gagaccgaaa atgaggctga gaacgaagtg gagaagacag acacccagca ggtggactgg
     2041 aactttgccc agtgcgaatt gctcgagaag caacgcattg atctaaaagc tgaaatgaag
     2101 aagcgtttag acaacttgga ggaacagtac aagcgggaga aacttcaggc cgatcagcaa
     2161 ttcgaggagc agcgcaaaac gtacgaggct cgcatcgatg ctttgcaaaa acaggttgag
     2221 gaacaatcca tgaccatgtc catgtacagc agctactcgc cggaggattt ccaccaggag
     2281 gaggacgtct acaccaaccc catgtacgaa tcctgctgga cagcaagaga ggctggcttg
     2341 gctgcctggg ctttccgcaa gtggcgttac caccaattca cctctttgcg agatgatctc
     2401 tggggcaatg ctatattcct taaggaagcc aatgccattt ccgttgagtt aaagaagaag
     2461 gtacaattcc aatttactct cttgaccgac accttgtact ctcctttgcc gcctgagctg
     2521 gcctccactg tggctcctgt gcatcaggag gatgagttcg gagctccacc tgtctctaag
     2581 accttggtgg ccgtcgaagt taccgatact aagaacggag ccactcatca ctggtccctg
     2641 gagaagttac ggcaacgttt ggagctgatg cgcgagatgt accacaacga agccgaaatg
     2701 agtcccactt caccggacta taatgtggag agcctcactg gtggagatcc attctacgat
     2761 cgcttcccct ggttccgcat ggtgggtcgc tccttcatct atctgagcaa cctgctctac
     2821 ccagtgccat tggtccacaa ggtggccatt gtcaacgaac gcggagatgt gcgtggctac
     2881 ctaaggattg ctgtgcagcc cgttctggat gaggagtcca ttgatttcaa taatggtgtc
     2941 aagcagtcag ctcgcttggt tttcaatgag gatgatgcca aaccgaaata ccgagctctc
     3001 aatgaaaagg atgatgtgca gcgctatatt gacaatggtg gtttggacag caagctggag
     3061 gaacttgagg atgtggactc tggtcgtggc attgactcta actccgcatc cgagtgccac
     3121 gaaaattcgg aggagcctgg cgagcatctg caggtgggca aggagttcac tttccgggtc
     3181 actgtgctac aggccactgg tattggggct gaatatgccg atatcttctg ccagttcaac
     3241 tttttgcatc gtcatgagga ggctttctcc accgaacccg tcaagaattc ggcatcgggt
     3301 gctcctctag gcttctacca tgtgcagaac ataactgtac ccgtaaccaa atccttcatc
     3361 gagtatttga agacccaacc cataatgttc aagatatttg ggcactacca gacgcaccca
     3421 ttgcataagg atgccaaaca ggattttgtt tctcggccac caccacgtcg catgttgcca
     3481 ccgagcatac cgatcagtca gccagtgcgt agtcccaagt ttggaccact accgtgtgca
     3541 cccacgtcta cggttttggc taagcacgat gttttggttt ggtttgaaat ctgtgaattg
     3601 gctcccaatg gagaatatgt gccatcggtg gtggagcaca gcgatgatct tccctgccgc
     3661 ggactgttcc tcttgcatca gggcatccag cggcgcattc gtattactat cgtacatgag
     3721 cccacaaccg aggtgaagtg gaaggacatc aatgagttgg tggttggacg tatccgcaat
     3781 acaccggagt catccgacga acaggacgaa gacgcctgtg tcttatcttt gggcttgttc
     3841 ccaggcgagg ctttggaggt gcccggagat gatagatctt tctaccgttt tgaggctgcc
     3901 tgggactcga gtctgcacaa ctcggcactg ctcaaccgcg tttctcaagg cggtgagacc
     3961 atctacatca ccctgagcgc ttacttggag ctggagaatt gcgcccgccc ggccataatt
     4021 accaaggacc tgagcatggt tatctatgga cgcgatgccc gcaccggtcc gcgctccctg
     4081 aagcacctgt tttcgggaca gtaccgcaat ccggaggcca atcgcctcac tggggtttac
     4141 gagctagcac tgcgcagagc atccgaagca ggtagtccag gtgtacaaag gcgtcaacgt
     4201 cgagtgctgg acaccagctc tacttatgtg cgcggcgagg agaatcttca tggctggagg
     4261 ccaaggggtg actccctgat cttcgatcac cagtgggagc tggagaaact caccagactt
     4321 gaagaggttg gacgcatgcg gcacttgctt ctgctgcgcg aacgtctggg catggacacc
     4381 aacccaaatc cgaccaccaa gaccgagaag gatgtttgca atctagctgc tcgggcagcc
     4441 acatcacccg tacatatggt cattccacaa tcgccgcaga ctccggtcaa ggatccacag
     4501 caaatcattc cagaacgcga atacaaccaa cgggagcagg atctcatgct taaatgctta
     4561 aagttggtgc agggacgcta tactaagagc gaggccaacg atacgcaaac tcagtcggat
     4621 gtttcgccta gcgatgaggg atgtgccgat atgaccgtca gctgcatctc cagcaattcc
     4681 atggaaaaca acaaatttgt aattcgacgc agattatgtt caccggatcg agctgatgca
     4741 cccaacgggt gggaggcacc tgctccggct actcagccag ctcttcccct ccgtctctat
     4801 gtgccggaac tggaggagat tcgcgtgagt cctgtggtgg cccgcaaggg tctgttgaat
     4861 gtcctggagc acggcggttc cggctggaag aagcgctggg tgatcgtccg tcgcccttac
     4921 gtgtttatct accgctcgga gaaggatccc gttgaacggg ctgtcctcaa tctggccact
     4981 gcgcatgtgg agtgcagcga ggaccaggcg gccatggtca agatacccaa cacattcagt
     5041 gtggtgacca agcatcgtgg ctatctgctg caaactctcg gtgacaagga agtacacgac
     5101 tggctgtatg ctatcaatcc attgcttgct gggcagataa aatcccgcct ggcgcgacgg
     5161 actttggagc ctgctagcca gacggcttcc cagatccagg ccaccaatgc ggcgaatgcc
     5221 aatagtgcga gcaaagggcg gccgctttcg aatatgagta aaggtgagga actctttaca
     5281 ggagttgtac ccatactcgt tgagttggac ggtgatgtaa atggtcataa gttttcggtc
     5341 aggggtgagg gcgagggtga cgcaactaac ggtaagctga ctttgaagtt tatctgtacc
     5401 acgggaaaac ttcccgtgcc gtggccaact ttggtaacaa cacttacata cggtgtgcaa
     5461 tgctttagtc gctacccgga tcacatgaaa aggcacgatt ttttcaaatc tgccatgcct
     5521 gagggttatg tgcaagagag aactatttcg ttcaaagacg acggaacgta caagacacgc
     5581 gccgaagtga aatttgaggg tgataccctt gtcaatcgta tagaacttaa aggaattgac
     5641 ttcaaggaag acggcaacat cttgggacac aaactggaat ataattttaa ctcgcacaac
     5701 gtgtatatca cagcagacaa acagaaaaac ggaatcaaag caaacttcaa aatacgccac
     5761 aacgttgaag atggaagcgt ccagctggct gatcattacc agcaaaacac ccccattggt
     5821 gatggaccgg tattgttgcc cgataatcat tatctctcaa cgcagtctgt actcagtaag
     5881 gatcctaacg aaaagaggga tcacatggtt cttctcgagt tcgtaaccgc agctggtatc
     5941 acgcacggca tggacgaact ctacaaaggc agtggcagtt ggtcgcatcc tcaattcgag
     6001 aagggcggcg gcagtggcgg cggcagtggc ggcagtgcct ggagtcatcc acaatttgag
     6061 aagtaaagcc tgcagtctcg acaagcttgt cgagaagtac tagaggatca taatcagcca
     6121 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     6181 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     6241 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6301 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatctgat cactgcttga
     6361 gcctagaaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     6421 ctgagcaata actatcataa cccctagggt atacccatct aattggaacc agataagtga
     6481 aatctagttc caaactattt tgtcattttt aattttcgta ttagcttacg acgctacacc
     6541 cagttcccat ctattttgtc actcttccct aaataatcct taaaaactcc atttccaccc
     6601 ctcccagttc ccaactattt tgtccgccca caaccggttg acttgggtca actgtcagac
     6661 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6721 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6781 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6841 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6901 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6961 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7021 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7081 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7141 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7201 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7261 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7321 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7381 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7441 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattgactt
     7501 gggtcgctct tcctgtggat gcgcagatgc cctgcgtaag cgggtgtggg cggacaataa
     7561 agtcttaaac tgaacaaaat agatctaaac tatgacaata aagtcttaaa ctagacagaa
     7621 tagttgtaaa ctgaaatcag tccagttatg ctgtgaaaaa gcatactgga cttttgttat
     7681 ggctaaagca aactcttcat tttctgaagt gcaaattgcc cgtcgtatta aagaggggcg
     7741 tggccaaggg catgtaaaga ctatattcgc ggcgttgtga caatttaccg aacaactccg
     7801 cggccgggaa gccgatctcg gcttgaacga attgttaggt ggcggtactt gggtcgatat
     7861 caaagtgcat cacttcttcc cgtatgccca actttgtata gagagccact gcgggatcgt
     7921 caccgtaatc tgcttgcacg tagatcacat aagcaccaag cgcgttggcc tcatgcttga
     7981 ggagattgat gagcgcggtg gcaatgccct gcctccggtg ctcgccggag actgcgagat
     8041 catagatata gatctcacta cgcggctgct caaacttggg cagaacgtaa gccgcgagag
     8101 cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat gaatgtctta ctacggagca
     8161 agttcccgag gtaatcggag tccggctgat gttgggagta ggtggctacg tctccgaact
     8221 cacgaccgaa aagatcaaga gcagcccgca tggatttgac ttggtcaggg ccgagcctac
     8281 atgtgcgaat gatgcccata cttgagccac ctaactttgt tttagggcga ctgccctgct
     8341 gcgtaacatc gttgctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc
     8401 acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacaaa aaaacagtca
     8461 taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg
     8521 gaccagttgc gtgagcgcat acgctacttg cattacagtt tacgaaccga acaggcttat
     8581 gtcaactggg ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc aaccttgggc
     8641 agcagcgaag tcgccataac ttcgtatagc atacattata cgaagttatc tgtaactat
//
