LOCUS       sequence-362698-        7829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human KIF5A(∆exon27) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7829)
  AUTHORS   Nakano J, Chiba K, Niwa S
  TITLE     An ALS-associated KIF5A mutant forms oligomers and aggregates and 
            induces neuronal toxicity.
  JOURNAL   Genes Cells. 2022 Apr 17. doi: 10.1111/gtc.12936.
  PUBMED    35430760
REFERENCE   2  (bases 1 to 7829)
  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: "Genes 
            Cells. 2022 Apr 17. doi: 10.1111/gtc.12936."
COMMENT     Sequence Label: pSN838
FEATURES             Location/Qualifiers
     source          1..7829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..441
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        442..645
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     596..616
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      684..693
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             690..1385
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(823..841)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1091..1110
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4694..4815
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4731..4750)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4785..4804
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4822..5277)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4959..4980)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5171..5190
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5304..5408
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5410..5767
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5410..5601
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5433..5453)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5618..5753
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5802..6596
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5802..6596
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5856..5875)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6466..6485
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6784..6803)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6828..6875
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7204..7792
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7693..7712
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
       61 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
      121 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      181 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      241 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      301 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      361 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      421 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      481 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      541 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      601 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
      661 cagatccgct agcgctaccg gtcgccacca tggtgagcaa gggcgaggca gtgatcaagg
      721 agttcatgcg gttcaaggtg cacatggagg gctccatgaa cggccacgag ttcgagatcg
      781 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
      841 agggtggccc cctgcccttc tcctgggaca tcctgtcccc tcagttcatg tacggctcca
      901 gggccttcac caagcacccc gccgacatcc ccgactacta taagcagtcc ttccccgagg
      961 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgccgtgacc gtgacccagg
     1021 acacctccct ggaggacggc accctgatct acaaggtgaa gctccgcggc accaacttcc
     1081 ctcctgacgg ccccgtaatg cagaagaaga caatgggctg ggaagcgtcc accgagcggt
     1141 tgtaccccga ggacggcgtg ctgaagggcg acattaagat ggccctgcgc ctgaaggacg
     1201 gcggccgcta cctggcggac ttcaagacca cctacaaggc caagaagccc gtgcagatgc
     1261 ccggcgccta caacgtcgac cgcaagttgg acatcacctc ccacaacgag gactacaccg
     1321 tggtggaaca gtacgaacgc tccgagggcc gccactccac cggcggcatg gacgagctgt
     1381 acaagtccgg actcagatct cgagctcaag cttcgaattc catggcggag accaacaacg
     1441 aatgtagcat caaggtgctc tgccgattcc ggcccctgaa ccaggctgag attctgcggg
     1501 gagacaagtt catccccatt ttccaagggg acgacagcgt cgttattggg gggaagccat
     1561 atgtttttga ccgtgtattc cccccaaaca cgactcaaga gcaagtttat catgcatgtg
     1621 ccatgcagat tgtcaaagat gtccttgctg gctacaatgg caccattttt gcttatggac
     1681 agacatcctc agggaaaaca cataccatgg agggaaagct gcacgaccct cagctgatgg
     1741 gaatcattcc tcgaattgcc cgagacatct tcaaccacat ctactccatg gatgagaacc
     1801 ttgagttcca catcaaggtt tcttactttg aaatttacct ggacaaaatt cgtgaccttc
     1861 tggatgtgac caagacaaat ctgtccgtgc acgaggacaa gaaccgggtg ccatttgtca
     1921 agggttgtac tgaacgcttt gtgtccagcc cggaggagat tctggatgtg attgatgaag
     1981 ggaaatcaaa tcgtcatgtg gctgtcacca acatgaatga acacagctct cggagccaca
     2041 gcatcttcct catcaacatc aagcaggaga acatggaaac ggagcagaag ctcagtggga
     2101 agctgtatct ggtggacctg gcagggagtg agaaggtcag caagactgga gcagagggag
     2161 ccgtgctgga cgaggcaaag aatatcaaca agtcactgtc agctctgggc aatgtgatct
     2221 ccgcactggc tgagggcact aaaagctatg ttccatatcg tgacagcaaa atgacaagga
     2281 ttctccagga ctctctcggg ggaaactgcc ggacgactat gttcatctgt tgctcaccat
     2341 ccagttataa tgatgcagag accaagtcca ccctgatgtt tgggcagcgg gcaaagacca
     2401 ttaagaacac tgcctcagta aatttggagt tgactgctga gcagtggaag aagaaatatg
     2461 agaaggagaa ggagaagaca aaggcccaga aggagacgat tgcgaagctg gaggctgagc
     2521 tgagccggtg gcgcaatgga gagaatgtgc ctgagacaga gcgcctggct ggggaggagg
     2581 cagccctggg agccgagctc tgtgaggaga cccctgtgaa tgacaactca tccatcgtgg
     2641 tgcgcatcgc gcccgaggag cggcagaaat acgaggagga gatccgccgt ctctataagc
     2701 agcttgacga caaggatgat gaaatcaacc aacaaagcca actcatagag aagctcaagc
     2761 agcaaatgct ggaccaggaa gagctgctgg tgtccacccg aggagacaac gagaaggtcc
     2821 agcgggagct gagccacctg caatcagaga acgatgccgc taaggatgag gtgaaggaag
     2881 tgctgcaggc cctggaggag ctggctgtga actatgacca gaagtcccag gaggtggagg
     2941 agaagagcca gcagaaccag cttctggtgg atgagctgtc tcagaaggtg gccaccatgc
     3001 tgtccctgga gtctgagttg cagcggctac aggaggtcag tggacaccag cgaaaacgaa
     3061 ttgctgaggt gctgaacggg ctgatgaagg atctgagcga gttcagtgtc attgtgggca
     3121 acggggagat taagctgcca gtggagatca gtggggccat cgaggaggag ttcactgtgg
     3181 cccgactcta catcagcaaa atcaaatcag aagtcaagtc tgtggtcaag cggtgccggc
     3241 agctggagaa cctccaggtg gagtgtcacc gcaagatgga agtgaccggg cgggagctct
     3301 catcctgcca gctcctcatc tctcagcatg aggccaagat ccgctcgctt acggaataca
     3361 tgcagagcgt ggagctaaag aagcggcacc tggaagagtc ctatgactcc ttgagcgatg
     3421 agctggccaa gctccaggcc caggaaactg tgcatgaagt ggccctgaag gacaaggagc
     3481 ctgacactca ggatgcagat gaagtgaaga aggctctgga gctgcagatg gagagtcacc
     3541 gggaggccca tcaccggcag ctggcccggc tccgggacga gatcaacgag aagcagaaga
     3601 ccattgatga gctcaaagac ctaaatcaga agctccagtt agagctagag aagcttcagg
     3661 ctgactacga gaagctgaag agcgaagaac acgagaagag caccaagctg caggagctga
     3721 catttctgta cgagcgacat gagcagtcca agcaggacct caagggtctg gaggagacag
     3781 ttgcccggga actccagacc ctccacaacc ttcgcaagct gttcgttcaa gacgtcacga
     3841 ctcgagtcaa gaaaagtgca gaaatggagc ccgaagacag tggggggatt cactcccaaa
     3901 agcagaagat ttcctttctt gagaacaacc tggaacagct tacaaaggtt cacaaacagc
     3961 tggtacgtga caatgcagat ctgcgttgtg agcttcctaa attggaaaaa cgacttaggg
     4021 ctacggctga gagagttaag gccctggagg gtgcactgaa ggaggccaag gagggcgcca
     4081 tgaaggacaa gcgccggtac cagcaggagg tggaccgcat caaggaggcc gttcgctaca
     4141 agagctcggg caaacggggc cattctgccc agattgccaa acccgtccgg cctggccact
     4201 acccagcatc ctcacccacc aacccctatg gcacccggag ccctgagtgc atcagttaca
     4261 ccaacagcct cttccagaac taccagaatc tctacctgca ggccacaccc agctccacct
     4321 cagatatgta ctttgcaaac tcctgtacca gcagtggagc cacatcttct ggcggcccct
     4381 tggcttccta ccagaaggcc aacatggaca atggagtgac ctgccgtgtg gctatgaggc
     4441 tgaggaccag gccaagcttt tccctctcca ccaagagaca gcagccagct aatctcccac
     4501 acccacggct gcatacctgc actttcagtt tctaagaggg actgaggcct cttctcagca
     4561 tgctggatcc accggatcta gataactgat cataatcagc cataccacat ttgtagaggt
     4621 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     4681 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4741 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4801 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     4861 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4921 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4981 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     5041 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     5101 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     5161 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     5221 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     5281 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5341 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5401 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     5461 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5521 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5581 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     5641 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     5701 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     5761 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     5821 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5881 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5941 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     6001 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     6061 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     6121 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     6181 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     6241 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     6301 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     6361 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     6421 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     6481 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     6541 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     6601 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     6661 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6721 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     6781 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     6841 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     6901 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     6961 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     7021 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     7081 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     7141 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7201 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     7261 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7321 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     7381 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7441 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7501 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7561 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     7621 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7681 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7741 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7801 caacgcggcc tttttacggt tcctggcct
//
