LOCUS       sequence-259350-        8798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of 6His-KIF1C-GFP in insect cells for protein 
            purification..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8798)
  AUTHORS   Siddiqui N, Zwetsloot AJ, Bachmann A, Roth D, Hussain H, Brandt J, 
            Kaverina I, Straube A
  TITLE     PTPN21 and Hook3 relieve KIF1C autoinhibition and activate 
            intracellular transport.
  JOURNAL   Nat Commun. 2019 Jun 19;10(1):2693. doi: 10.1038/s41467-019-10644-9.
  PUBMED    31217419
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"; journalName: "Nat 
            Commun."; date: "2019-06-19"; volume: "10(1):2693. doi"; pages: " 
            10.1038/s41467-019-10644-9"
COMMENT     Sequence Label: pFastBac-M13-6His-KIF1C-GFP
FEATURES             Location/Qualifiers
     source          1..8798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        482..586
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(587..655,656..1447)
                     /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(805..824)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1618..2206
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2107..2126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2360..2377
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2511..2735
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2802..3335)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3505..3510,3511..3527,3528..3533))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3885..3976
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3901..3918
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4041..4058
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4080..4100
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     regulatory      6342..6351
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      7440..7449
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7446..7448,7449..7451,7452..8165)
                     /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(7446..7448,7449..7451,7452..8162)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7491..7512)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7752..7771)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8099..8120
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    8288..8422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8325..8344)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8379..8398
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  8451..8616
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
      481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      541 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
      601 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
      661 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      721 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      781 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      841 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      901 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      961 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1021 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1081 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1141 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1201 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1261 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1321 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     1381 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     1441 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     1501 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     1561 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1621 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1681 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1741 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1801 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1861 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1921 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1981 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2041 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2101 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2161 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2221 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     2281 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2341 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     2401 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct
     2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2941 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3301 ccctgctgcg taacatcgtt gctgctgcgt aacatcaaac atcgacccac ggcgtaacgc
     3361 gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg
     3421 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     3481 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt
     3541 cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc
     3601 gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag
     3661 gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct gccctggctt
     3721 caggagatcg gaagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa
     3781 gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc
     3841 tatagttcta gtggttggct acgtatactc cggaatatta atagatcatg gagataatta
     3901 aaatgataac catctcgcaa ataaataagt attttactgt tttcgtaaca gttttgtaat
     3961 aaaaaaacct ataaatattc cggattattc atacccccac catcgggcgc ggatctcggt
     4021 ccgaaaccat gtcgtactac catcaccatc accatcacga ttacgatatc ccaacgaccg
     4081 aaaacctgta ttttcagggc gccatggcgc gcccggatct ggctggtgcc tcggtgaaag
     4141 tggcagtgag ggttcggccc tttaacgccc gtgagaccag ccaggatgcc aagtgtgtgg
     4201 tcagcatgca gggcaacacc acctccatca tcaatcctaa acagagcaag gatgccccca
     4261 aaagcttcac ctttgactac tcctactggt cacacacttc gacggaggac ccccagtttg
     4321 catctcagca gcaagtgtat cgggacatcg gagaagagat gctgctccac gcctttgaag
     4381 gctacaacgt gtgcatcttt gcctatgggc agaccggggc tgggaaatcc tataccatga
     4441 tggggcgaca ggagccaggg cagcagggca tcgtgcccca gctctgtgag gacctcttct
     4501 ctcgcgttag tgagaaccag agtgctcagc tatcctactc tgtggaggtg agctatatgg
     4561 agatctactg tgagcgggta cgagacctct tgaaccccaa gagtcggggt tctctgcggg
     4621 tccgggagca ccccatcctg ggcccgtacg tgcaggacct gtccaaattg gctgtgacct
     4681 cctacgcaga cattgctgac ctcatggact gtggaaataa agcacggact gtggctgcca
     4741 ccaacatgaa tgagaccagc agccgttccc atgccgtctt taccatcgtc ttcacacagc
     4801 gctgccatga ccagctcacg gggctggact cggagaaggt cagtaagatc agtttggtgg
     4861 accttgctgg gagtgagcga gccgactcct caggggcccg gggcatgcgc ctgaaggaag
     4921 gagccaacat caataagtcc ctgactacac tagggaaagt gatctcggcc cttgcagata
     4981 tgcaatcaaa gaagcgaaag tcggatttta tcccctacag ggactctgtg ctcacctggc
     5041 tgctcaagga aaatttgggg gggaactcac gcacagccat gattgcagcc ctgagccctg
     5101 ctgacatcaa ttacgaggag actctcagca ccctcaggta tgctgaccgc accaagcaaa
     5161 tccgctgcaa tgccatcatc aacgaggacc ctaatgcccg gctgattaga gagctgcagg
     5221 aggaagtagc ccggctgcgg gaactgctga tggctcaggg actgtcagcc tctgctctgg
     5281 aaggcctgaa gacggaagaa gggagtgtca gaggcgccct gccagctgtg tcatctcccc
     5341 cagctccagt ttcaccctca tcacccacca cacataatgg ggagctggag ccgtcattct
     5401 cccccaacac ggagtcccag attgggcctg aggaagccat ggagaggctg caggagacag
     5461 agaagattat agctgagctg aacgagacat gggaggagaa gctacgcaag acagaagccc
     5521 tgaggatgga gagagaagca ttgctggctg agatgggggt ggccgtccgg gaggatgggg
     5581 gaactgtggg cgtcttctct ccaaagaaga ctccccacct ggtgaacctg aacgaagacc
     5641 ctctgatgtc tgagtgtctg ctctaccaca tcaaagatgg cgtcaccagg gtcggccaag
     5701 tagatatgga catcaagctg accggacagt tcattcggga gcaacactgt ctgttccgga
     5761 gcatccccca gccagatgga gaagtggtgg tcactctgga gccttgtgaa ggagctgaga
     5821 catatgtgaa tgggaagctt gtgacggagc cgctggtgct gaagtcaggg aataggattg
     5881 tgatgggcaa gaaccacgtt ttccgcttca accacccgga gcaggcaagg ctggaacggg
     5941 aacgaggggt ccccccaccc ccaggaccgc cctctgagcc agtcgactgg aactttgccc
     6001 agaaggaact gctggagcag caaggcatcg acataaagct ggaaatggag aagaggctgc
     6061 aggatctgga gaatcagtac cggaaagaaa aggaagaagc cgatcttctg ctggagcagc
     6121 agcgactgta tgcagactcg gacagcgggg atgactctga caagcgctct tgtgaagaga
     6181 gctggaggct catctcctcc ttgcgggagc agctgccgcc caccacggtc cagaccattg
     6241 tcaaacgctg tggtctgccc agcagtggca agcgcagggc ccctcgcagg gtttatcaga
     6301 tcccccagcg acgcaggctg cagggcaaag acccccgctg ggccaccatg gctgacctga
     6361 agatgcaggc ggtgaaggag atctgctacg aggtggccct ggctgacttc cgccacgggc
     6421 gggctgagat tgaggccctg gccgccctca agatgcggga gctgtgtcgc acctatggca
     6481 agccagacgg ccccggagac gcctggaggg ctgtggcccg ggatgtctgg gacactgtag
     6541 gcgaggagga aggaggtgga gctggcagtg gtggtggcag tgaggaggga gcccgagggg
     6601 cggaggtgga ggacctccgg gcccacatcg acaagctgac ggggattctg caggaggtga
     6661 agctgcagaa cagcagcaag gaccgggagc tgcaggccct gcgggaccgc atgctccgca
     6721 tggagagggt catccccctg gcccaggatc atgaggatga gaatgaagaa ggtggtgagg
     6781 tcccctgggc cccgcctgaa ggatcagagg cagcagagga ggcagccccc agtgaccgca
     6841 tgccgtcagc ccggcccccc tcgccaccac tgtcaagctg ggagcgggtg tcacggctca
     6901 tggaggagga ccctgccttc cgtcgtggtc gtcttcgctg gctcaagcag gagcagctac
     6961 ggctgcaggg actgcagggc tctgggggcc ggggcggggg gctgcgcagg cccccagccc
     7021 gctttgtgcc ccctcacgac tgcaagctac gcttcccctt caagagcaac ccccagcacc
     7081 gggagtcttg gccagggatg gggagcgggg aggctccaac tccgctccaa ccccctgagg
     7141 aggtcactcc ccatccagcc acccctgccc gccggcctcc gagtccccga aggtcccacc
     7201 atccccgcag gaactccctg gatggagggg gccgatcccg gggagcgggt tctgcacagc
     7261 ctgaacccca gcacttccag cccaaaaagc acaactctta tccccagcca ccccaaccct
     7321 acccagccca gcggccccca gggccccgct accccccata cactactccc ccacgaatga
     7381 gacggcagcg ttctgcccct gacctcaagg agagtggggc agctgtggat ccaccggtcg
     7441 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     7501 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     7561 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     7621 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     7681 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     7741 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     7801 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     7861 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     7921 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     7981 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     8041 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     8101 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     8161 agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     8221 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     8281 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     8341 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     8401 tgtatcttat catgtctgga tctgatcact gatatcgcct aggagatccg aaccagataa
     8461 gtgaaatcta gttccaaact attttgtcat ttttaatttt cgtattagct tacgacgcta
     8521 cacccagttc ccatctattt tgtcactctt ccctaaataa tccttaaaaa ctccatttcc
     8581 acccctccca gttcccaact attttgtccg cccacagcgg ggcatttttc ttcctgttat
     8641 gtttttaatc aaacatcctg ccaactccat gtgacaaacc gtcatcttcg gctacttttt
     8701 ctctgtcaca gaatgaaaat ttttctgtca tctcttcgtt attaatgttt gtaattgact
     8761 gaatatcaac gcttatttgc agcctgaatg gcgaatgg
//
