LOCUS       sequence-362697-        7857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human KIF5A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7857)
  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 7857)
  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: pSN837
FEATURES             Location/Qualifiers
     source          1..7857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        136..439
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        440..643
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     594..614
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      682..691
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             688..1383
                     /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(821..839)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1089..1108
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4720..4841
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4757..4776)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4811..4830
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4848..5303)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4985..5006)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5197..5216
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5330..5434
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5436..5793
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5436..5627
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5459..5479)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5644..5779
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5828..6622
                     /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             5828..6622
                     /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(5882..5901)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6492..6511
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6810..6829)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6854..6901
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7230..7818
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7719..7738
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
       61 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc
      121 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      181 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      421 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      481 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      541 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      601 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      661 gatccgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggcagt gatcaaggag
      721 ttcatgcggt tcaaggtgca catggagggc tccatgaacg gccacgagtt cgagatcgag
      781 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
      841 ggtggccccc tgcccttctc ctgggacatc ctgtcccctc agttcatgta cggctccagg
      901 gccttcacca agcaccccgc cgacatcccc gactactata agcagtcctt ccccgagggc
      961 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg ccgtgaccgt gacccaggac
     1021 acctccctgg aggacggcac cctgatctac aaggtgaagc tccgcggcac caacttccct
     1081 cctgacggcc ccgtaatgca gaagaagaca atgggctggg aagcgtccac cgagcggttg
     1141 taccccgagg acggcgtgct gaagggcgac attaagatgg ccctgcgcct gaaggacggc
     1201 ggccgctacc tggcggactt caagaccacc tacaaggcca agaagcccgt gcagatgccc
     1261 ggcgcctaca acgtcgaccg caagttggac atcacctccc acaacgagga ctacaccgtg
     1321 gtggaacagt acgaacgctc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     1381 aagtccggac tcagatctcg agctcaagct tcgaattcca tggcggagac caacaacgaa
     1441 tgtagcatca aggtgctctg ccgattccgg cccctgaacc aggctgagat tctgcgggga
     1501 gacaagttca tccccatttt ccaaggggac gacagcgtcg ttattggggg gaagccatat
     1561 gtttttgacc gtgtattccc cccaaacacg actcaagagc aagtttatca tgcatgtgcc
     1621 atgcagattg tcaaagatgt ccttgctggc tacaatggca ccatttttgc ttatggacag
     1681 acatcctcag ggaaaacaca taccatggag ggaaagctgc acgaccctca gctgatggga
     1741 atcattcctc gaattgcccg agacatcttc aaccacatct actccatgga tgagaacctt
     1801 gagttccaca tcaaggtttc ttactttgaa atttacctgg acaaaattcg tgaccttctg
     1861 gatgtgacca agacaaatct gtccgtgcac gaggacaaga accgggtgcc atttgtcaag
     1921 ggttgtactg aacgctttgt gtccagcccg gaggagattc tggatgtgat tgatgaaggg
     1981 aaatcaaatc gtcatgtggc tgtcaccaac atgaatgaac acagctctcg gagccacagc
     2041 atcttcctca tcaacatcaa gcaggagaac atggaaacgg agcagaagct cagtgggaag
     2101 ctgtatctgg tggacctggc agggagtgag aaggtcagca agactggagc agagggagcc
     2161 gtgctggacg aggcaaagaa tatcaacaag tcactgtcag ctctgggcaa tgtgatctcc
     2221 gcactggctg agggcactaa aagctatgtt ccatatcgtg acagcaaaat gacaaggatt
     2281 ctccaggact ctctcggggg aaactgccgg acgactatgt tcatctgttg ctcaccatcc
     2341 agttataatg atgcagagac caagtccacc ctgatgtttg ggcagcgggc aaagaccatt
     2401 aagaacactg cctcagtaaa tttggagttg actgctgagc agtggaagaa gaaatatgag
     2461 aaggagaagg agaagacaaa ggcccagaag gagacgattg cgaagctgga ggctgagctg
     2521 agccggtggc gcaatggaga gaatgtgcct gagacagagc gcctggctgg ggaggaggca
     2581 gccctgggag ccgagctctg tgaggagacc cctgtgaatg acaactcatc catcgtggtg
     2641 cgcatcgcgc ccgaggagcg gcagaaatac gaggaggaga tccgccgtct ctataagcag
     2701 cttgacgaca aggatgatga aatcaaccaa caaagccaac tcatagagaa gctcaagcag
     2761 caaatgctgg accaggaaga gctgctggtg tccacccgag gagacaacga gaaggtccag
     2821 cgggagctga gccacctgca atcagagaac gatgccgcta aggatgaggt gaaggaagtg
     2881 ctgcaggccc tggaggagct ggctgtgaac tatgaccaga agtcccagga ggtggaggag
     2941 aagagccagc agaaccagct tctggtggat gagctgtctc agaaggtggc caccatgctg
     3001 tccctggagt ctgagttgca gcggctacag gaggtcagtg gacaccagcg aaaacgaatt
     3061 gctgaggtgc tgaacgggct gatgaaggat ctgagcgagt tcagtgtcat tgtgggcaac
     3121 ggggagatta agctgccagt ggagatcagt ggggccatcg aggaggagtt cactgtggcc
     3181 cgactctaca tcagcaaaat caaatcagaa gtcaagtctg tggtcaagcg gtgccggcag
     3241 ctggagaacc tccaggtgga gtgtcaccgc aagatggaag tgaccgggcg ggagctctca
     3301 tcctgccagc tcctcatctc tcagcatgag gccaagatcc gctcgcttac ggaatacatg
     3361 cagagcgtgg agctaaagaa gcggcacctg gaagagtcct atgactcctt gagcgatgag
     3421 ctggccaagc tccaggccca ggaaactgtg catgaagtgg ccctgaagga caaggagcct
     3481 gacactcagg atgcagatga agtgaagaag gctctggagc tgcagatgga gagtcaccgg
     3541 gaggcccatc accggcagct ggcccggctc cgggacgaga tcaacgagaa gcagaagacc
     3601 attgatgagc tcaaagacct aaatcagaag ctccagttag agctagagaa gcttcaggct
     3661 gactacgaga agctgaagag cgaagaacac gagaagagca ccaagctgca ggagctgaca
     3721 tttctgtacg agcgacatga gcagtccaag caggacctca agggtctgga ggagacagtt
     3781 gcccgggaac tccagaccct ccacaacctt cgcaagctgt tcgttcaaga cgtcacgact
     3841 cgagtcaaga aaagtgcaga aatggagccc gaagacagtg gggggattca ctcccaaaag
     3901 cagaagattt cctttcttga gaacaacctg gaacagctta caaaggttca caaacagctg
     3961 gtacgtgaca atgcagatct gcgttgtgag cttcctaaat tggaaaaacg acttagggct
     4021 acggctgaga gagttaaggc cctggagggt gcactgaagg aggccaagga gggcgccatg
     4081 aaggacaagc gccggtacca gcaggaggtg gaccgcatca aggaggccgt tcgctacaag
     4141 agctcgggca aacggggcca ttctgcccag attgccaaac ccgtccggcc tggccactac
     4201 ccagcatcct cacccaccaa cccctatggc acccggagcc ctgagtgcat cagttacacc
     4261 aacagcctct tccagaacta ccagaatctc tacctgcagg ccacacccag ctccacctca
     4321 gatatgtact ttgcaaactc ctgtaccagc agtggagcca catcttctgg cggccccttg
     4381 gcttcctacc agaaggccaa catggacaat ggaaatgcca cagatatcaa tgacaatagg
     4441 agtgacctgc cgtgtggcta tgaggctgag gaccaggcca agcttttccc tctccaccaa
     4501 gagacagcag ccagctaatc tcccacaccc acggctgcat acctgcactt tcagtttcta
     4561 agagggactg aggcctcttc tcagcatgct ggatccaccg gatctagata actgatcata
     4621 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     4681 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     4741 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4801 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     4861 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     4921 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     4981 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     5041 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     5101 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     5161 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     5221 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     5281 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5341 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5401 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     5461 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5521 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     5581 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5641 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5701 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     5761 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     5821 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     5881 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     5941 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     6001 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     6061 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     6121 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     6181 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     6241 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     6301 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     6361 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     6421 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     6481 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     6541 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     6601 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     6661 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6721 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     6781 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     6841 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     6901 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     6961 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     7021 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     7081 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7141 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7201 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7261 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7321 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     7381 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7441 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7501 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7561 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7621 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7681 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7741 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7801 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttt
//
