LOCUS       sequence-449869-        8699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Drosophila UNC-104(A255V) 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: pKC51
FEATURES             Location/Qualifiers
     source          1..8699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      190..778
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     679..698
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  913..1136
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1203..1736)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1925..1930,1931..1947,1948..1953))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(2024..2057)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2134..2225
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2150..2167
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             join(7321..7323,7324..8034)
                     /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(8047..8070,8071..8106,8107..8130)
                     /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    8279..8413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8316..8335)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8370..8389
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  8534..8699
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 accggttgac ttgggtcaac tgtcagacca agtttactca tatatacttt agattgattt
       61 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
      121 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
      181 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
      241 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      301 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
      361 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      421 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      481 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      541 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      601 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      661 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      721 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      781 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      841 ctttcctgcg ttatcccctg attgacttgg gtcgctcttc ctgtggatgc gcagatgccc
      901 tgcgtaagcg ggtgtgggcg gacaataaag tcttaaactg aacaaaatag atctaaacta
      961 tgacaataaa gtcttaaact agacagaata gttgtaaact gaaatcagtc cagttatgct
     1021 gtgaaaaagc atactggact tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc
     1081 aaattgcccg tcgtattaaa gaggggcgtg gccaagggca tgtaaagact atattcgcgg
     1141 cgttgtgaca atttaccgaa caactccgcg gccgggaagc cgatctcggc ttgaacgaat
     1201 tgttaggtgg cggtacttgg gtcgatatca aagtgcatca cttcttcccg tatgcccaac
     1261 tttgtataga gagccactgc gggatcgtca ccgtaatctg cttgcacgta gatcacataa
     1321 gcaccaagcg cgttggcctc atgcttgagg agattgatga gcgcggtggc aatgccctgc
     1381 ctccggtgct cgccggagac tgcgagatca tagatataga tctcactacg cggctgctca
     1441 aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca
     1501 agcgcgatga atgtcttact acggagcaag ttcccgaggt aatcggagtc cggctgatgt
     1561 tgggagtagg tggctacgtc tccgaactca cgaccgaaaa gatcaagagc agcccgcatg
     1621 gatttgactt ggtcagggcc gagcctacat gtgcgaatga tgcccatact tgagccacct
     1681 aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctgcg taacatcgtt
     1741 gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct tggatgcccg
     1801 aggcatagac tgtacaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt
     1861 accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca
     1921 ttacagttta cgaaccgaac aggcttatgt caactgggtt cgtgccttca tccgtttcca
     1981 cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc gccataactt cgtatagcat
     2041 acattatacg aagttatctg taactataac ggtcctaagg tagcgagttt aaacactagt
     2101 atcgattcgc gacctactcc ggaatattaa tagatcatgg agataattaa aatgataacc
     2161 atctcgcaaa taaataagta ttttactgtt ttcgtaacag ttttgtaata aaaaaaccta
     2221 taaatattcc ggattattca taccgtccca ccatcgggcg cggatcccgg tccgaagcgc
     2281 gcggaattca tgtcgtcggt taaggtggcg gtgcgagtgc gccccttcaa ctcgcgcgaa
     2341 atagccaggg agtcgaaatg catcatcgag atggcaggag ccacaacggc catcaccaat
     2401 ccaaaggtgc cgcccaacac aagcgattca gtaaagcgct tcaactttga ttactcctac
     2461 tggtcacatg atcaccacga tgccgatttc tctacacaat cgatggtgta caaggacatt
     2521 ggcgaggaga tgttacagca ctccttcgat ggctacaatg tctgtatctt tgcctatggc
     2581 cagactggag ctggaaagtc gtataccatg atgggcaggc aggaggagca gcaagagggc
     2641 ataataccaa tgatttgcaa ggatctgttc actcgcatac aggatactga gaccgatgac
     2701 ctcaagtact cggttgaggt ttcctatatg gaaatctatt gcgagcgagt cagggatttg
     2761 ttgaatccga aaaacaaggg caatctgcgt gtgagggagc atcccctttt gggtccctat
     2821 gtggaggatt tgtccaaact agccgttacc gattaccagg acattcacga tctcattgat
     2881 gaaggcaaca aagctcgaac tgtggcagct actaacatga acgaaaccag ttcccgctct
     2941 catgccgtct ttacaatctt ctttacccaa cgtcgtcatg atctgatgac aaatttgacc
     3001 acagaaaagg tatccaagat aagcttggta gacttggccg ggtcggaacg agtagattcc
     3061 actggtgcca agggcactcg cttgaaggag ggagccaaca ttaacaagtc cttgaccaca
     3121 ttgggcaaag ttatctcagc tttggcggaa gtgtctgctt ccaagaaaaa gaacaccaag
     3181 aaggcagatt ttattccgta ccgtgattcg gccttgacct ggctgttacg tgaaaacttg
     3241 ggaggaaatt cgaagacagc tatgattgcg gctatctcac cagcagatat taactacgat
     3301 gaaaccctca gcacactgcg ctatgcggat cgtgccaagc aaattgtttg caaggctgtt
     3361 gtcaatgaag atgccaatgc caagctgatt cgagaactca aggaggagat ccaaaagctg
     3421 cgggacttac tcaaagccga gggcattgaa gtgcaggaag aggatgagct caccaagtca
     3481 acggtgatca agtcgcccac taagtcacgt aatcgtaatg gatccaccac ggagatggct
     3541 gtggatcagc tgcaggccag cgagaaactt attgcagagc tcaacgagac ctgggaggag
     3601 aaacttaagc gcaccgagga gattcgcgtg cagcgagagg cggtcttcgc cgaaatgggt
     3661 gtggctgtca aggaagatgg aataacagtt ggcgtattct caccgaagaa gactccgcat
     3721 ttggtcaacc taaacgagga tcccaatctg tctgagtgtc tgctttacta catcaaggag
     3781 ggtctaactc ggttgggtac ccatgaagca aatgtgcccc aggacattca gctctccgga
     3841 tcgcacatcc tcaaggagca ctgcaccttt gagaacaaga acagcacggt tacattgctg
     3901 ccacacaagg atgctatcat ctatgtaaat ggacgcaagt tggttgaacc ggaggttctt
     3961 aagaccggtt ctcgcgtgat cctcggaaag aaccacgtgt tccgctttac caatccagaa
     4021 caggcacgcg aattacggga taagatcgag accgaaaatg aggctgagaa cgaagtggag
     4081 aagacagaca cccagcaggt ggactggaac tttgcccagt gcgaattgct cgagaagcaa
     4141 ggcattgatc taaaagctga aatgaagaag cgtttagaca acttggagga acagtacaag
     4201 cgggagaaac ttcaggccga tcagcaattc gaggagcagc gcaaaacgta cgaggctcgc
     4261 atcgatgctt tgcaaaaaca ggttgaggaa caatccatga ccatgtccat gtacagcagc
     4321 tactcgccgg aggatttcca ccaggaggag gacgtctaca ccaaccccat gtacgaatcc
     4381 tgctggacag caagagaggc tggcttggct gcctgggctt tccgcaagtg gcgttaccac
     4441 caattcacct ctttgcgaga tgatctctgg ggcaatgcta tattccttaa ggaagccaat
     4501 gccatttccg ttgagttaaa gaagaaggta caattccaat ttactctctt gaccgacacc
     4561 ttgtactctc ctttgccgcc tgagctggcc tccactgtgg ctcctgtgca tcaggaggat
     4621 gagttcggag ctccacctgt ctctaagacc ttggtggccg tcgaagttac cgatactaag
     4681 aacggagcca ctcatcactg gtccctggag aagttacggc aacgtttgga gctgatgcgc
     4741 gagatgtacc acaacgaagc cgaaatgagt cccacttcac cggactataa tgtggagagc
     4801 ctcactggtg gagatccatt ctacgatcgc ttcccctggt tccgcatggt gggtcgctcc
     4861 ttcatctatc tgagcaacct gctctaccca gtgccattgg tccacaaggt ggccattgtc
     4921 aacgaacgcg gagatgtgcg tggctaccta aggattgctg tgcagcccgt tctggatgag
     4981 gagtccattg atttcaataa tggtgtcaag cagtcagctc gcttggtttt caatgaggat
     5041 gatgccaaac cgaaataccg agctctcaat gaaaaggatg atgtgcagcg ctatattgac
     5101 aatggtggtt tggacagcaa gctggaggaa cttgaggatg tggactctgg tcgtggcatt
     5161 gactctaact ccgcatccga gtgccacgaa aattcggagg agcctggcga gcatctgcag
     5221 gtgggcaagg agttcacttt ccgggtcact gtgctacagg ccactggtat tggggctgaa
     5281 tatgccgata tcttctgcca gttcaacttt ttgcatcgtc atgaggaggc tttctccacc
     5341 gaacccgtca agaattcggc atcgggtgct cctctaggct tctaccatgt gcagaacata
     5401 actgtacccg taaccaaatc cttcatcgag tatttgaaga cccaacccat aatgttcaag
     5461 atatttgggc actaccagac gcacccattg cataaggatg ccaaacagga ttttgtttct
     5521 cggccaccac cacgtcgcat gttgccaccg agcataccga tcagtcagcc agtgcgtagt
     5581 cccaagtttg gaccactacc gtgtgcaccc acgtctacgg ttttggctaa gcacgatgtt
     5641 ttggtttggt ttgaaatctg tgaattggct cccaatggag aatatgtgcc atcggtggtg
     5701 gagcacagcg atgatcttcc ctgccgcgga ctgttcctct tgcatcaggg catccagcgg
     5761 cgcattcgta ttactatcgt acatgagccc acaaccgagg tgaagtggaa ggacatcaat
     5821 gagttggtgg ttggacgtat ccgcaataca ccggagtcat ccgacgaaca ggacgaagac
     5881 gcctgtgtct tatctttggg cttgttccca ggcgaggctt tggaggtgcc cggagatgat
     5941 agatctttct accgttttga ggctgcctgg gactcgagtc tgcacaactc ggcactgctc
     6001 aaccgcgttt ctcaaggcgg tgagaccatc tacatcaccc tgagcgctta cttggagctg
     6061 gagaattgcg cccgcccggc cataattacc aaggacctga gcatggttat ctatggacgc
     6121 gatgcccgca ccggtccgcg ctccctgaag cacctgtttt cgggacagta ccgcaatccg
     6181 gaggccaatc gcctcactgg ggtttacgag ctagcactgc gcagagcatc cgaagcaggt
     6241 agtccaggtg tacaaaggcg tcaacgtcga gtgctggaca ccagctctac ttatgtgcgc
     6301 ggcgaggaga atcttcatgg ctggaggcca aggggtgact ccctgatctt cgatcaccag
     6361 tgggagctgg agaaactcac cagacttgaa gaggttggac gcatgcggca cttgcttctg
     6421 ctgcgcgaac gtctgggcat ggacaccaac ccaaatccga ccaccaagac cgagaaggat
     6481 gtttgcaatc tagctgctcg ggcagccaca tcacccgtac atatggtcat tccacaatcg
     6541 ccgcagactc cggtcaagga tccacagcaa atcattccag aacgcgaata caaccaacgg
     6601 gagcaggatc tcatgcttaa atgcttaaag ttggtgcagg gacgctatac taagagcgag
     6661 gccaacgata cgcaaactca gtcggatgtt tcgcctagcg atgagggatg tgccgatatg
     6721 accgtcagct gcatctccag caattccatg gaaaacaaca aatttgtaat tcgacgcaga
     6781 ttatgttcac cggatcgagc tgatgcaccc aacgggtggg aggcacctgc tccggctact
     6841 cagccagctc ttcccctccg tctctatgtg ccggaactgg aggagattcg cgtgagtcct
     6901 gtggtggccc gcaagggtct gttgaatgtc ctggagcacg gcggttccgg ctggaagaag
     6961 cgctgggtga tcgtccgtcg cccttacgtg tttatctacc gctcggagaa ggatcccgtt
     7021 gaacgggctg tcctcaatct ggccactgcg catgtggagt gcagcgagga ccaggcggcc
     7081 atggtcaaga tacccaacac attcagtgtg gtgaccaagc atcgtggcta tctgctgcaa
     7141 actctcggtg acaaggaagt acacgactgg ctgtatgcta tcaatccatt gcttgctggg
     7201 cagataaaat cccgcctggc gcgacggact ttggagcctg ctagccagac ggcttcccag
     7261 atccaggcca ccaatgcggc gaatgccaat agtgcgagca aagggcggcc gctttcgaat
     7321 atgagtaaag gtgaggaact ctttacagga gttgtaccca tactcgttga gttggacggt
     7381 gatgtaaatg gtcataagtt ttcggtcagg ggtgagggcg agggtgacgc aactaacggt
     7441 aagctgactt tgaagtttat ctgtaccacg ggaaaacttc ccgtgccgtg gccaactttg
     7501 gtaacaacac ttacatacgg tgtgcaatgc tttagtcgct acccggatca catgaaaagg
     7561 cacgattttt tcaaatctgc catgcctgag ggttatgtgc aagagagaac tatttcgttc
     7621 aaagacgacg gaacgtacaa gacacgcgcc gaagtgaaat ttgagggtga tacccttgtc
     7681 aatcgtatag aacttaaagg aattgacttc aaggaagacg gcaacatctt gggacacaaa
     7741 ctggaatata attttaactc gcacaacgtg tatatcacag cagacaaaca gaaaaacgga
     7801 atcaaagcaa acttcaaaat acgccacaac gttgaagatg gaagcgtcca gctggctgat
     7861 cattaccagc aaaacacccc cattggtgat ggaccggtat tgttgcccga taatcattat
     7921 ctctcaacgc agtctgtact cagtaaggat cctaacgaaa agagggatca catggttctt
     7981 ctcgagttcg taaccgcagc tggtatcacg cacggcatgg acgaactcta caaaggcagt
     8041 ggcagttggt cgcatcctca attcgagaag ggcggcggca gtggcggcgg cagtggcggc
     8101 agtgcctgga gtcatccaca atttgagaag taaagcctgc agtctcgaca agcttgtcga
     8161 gaagtactag aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     8221 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     8281 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     8341 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     8401 tcatgtctgg atctgatcac tgcttgagcc tagaagatcc ggctgctaac aaagcccgaa
     8461 aggaagctga gttggctgct gccaccgctg agcaataact atcataaccc ctagggtata
     8521 cccatctaat tggaaccaga taagtgaaat ctagttccaa actattttgt catttttaat
     8581 tttcgtatta gcttacgacg ctacacccag ttcccatcta ttttgtcact cttccctaaa
     8641 taatccttaa aaactccatt tccacccctc ccagttccca actattttgt ccgcccaca
//
