LOCUS       sequence-328016-        8537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged KIF5A R280H protein in pcDNA3.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8537)
  AUTHORS   Fukuda Y, Pazyra-Murphy MF, Silagi ES, Tasdemir-Yilmaz OE, Li Y, 
            Rose L, Yeoh ZC, Vangos NE, Geffken EA, Seo HS, Adelmant G, Bird GH,
            Walensky LD, Marto JA, Dhe-Paganon S, Segal RA
  TITLE     Binding and transport of SFPQ-RNA granules by KIF5A/KLC1 motors 
            promotes axon survival.
  JOURNAL   J Cell Biol. 2021 Jan 4;220(1). pii: 211577. doi: 
            10.1083/jcb.202005051.
  PUBMED    33284322
REFERENCE   2  (bases 1 to 8537)
  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 
            Biol."; date: "2021-01-4"; volume: "220(1). pii: 211577. doi"; 
            pages: " 10.1083/jcb.202005051"
COMMENT     Sequence Label: FLAG-KIF5AR280H
FEATURES             Location/Qualifiers
     source          1..8537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             708..731
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3894..3911)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3900..4124
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4170..4598
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4257..4276)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4467..4488
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4607..4627)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4612..4941
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4792..4927
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4854..4873
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5008..5802
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5062..5081)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5672..5691
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5976..6097
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6013..6032)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6067..6086
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6146..6162)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6146..6162)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6159..6181)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6170..6186
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6194..6200,6201..6218,6219..6224))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6239..6260
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6377..6394)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6548..7136)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6628..6647)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7307..8098,8099..8167))
                     /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     7930..7949
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8168..8272)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8347..8366)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc caccatggac tacaaagacg
      721 atgacgacaa gggatccgga atggcggaga ctaacaacga atgcagcatc aaggtgcttt
      781 gccgatttcg gcccctgaac caggccgaga ttctgcgggg ggacaagttc atccccattt
      841 tccaagggga cgacagcgtc attattgggg gaaagccata tgtctttgac cgcgtcttcc
      901 ccccaaacac cactcaggag caggtttacc acgcctgtgc catgcagatc gtcaaagacg
      961 tccttgctgg ttacaatggc acaatcttcg cttatggaca gacatcctca gggaaaacgc
     1021 ataccatgga ggggaagctg cacgaccctc agctgatggg catcattccc cggatcgctc
     1081 gagacatctt caaccacatc tactccatgg atgagaacct tgaattccac attaaggtat
     1141 cttacttcga gatttacctg gataagatcc gtgacctttt ggatgtgacc aagacgaacc
     1201 tgtccgtgca tgaggacaaa aaccgggtgc cgtttgtcaa gggttgtacc gaacgctttg
     1261 tgtccagccc agaggagatt ctggatgtga tcgatgaggg gaagtccaac cgtcacgtag
     1321 ctgtcaccaa catgaacgag cacagttctc ggagccacag catcttcctc atcaacatca
     1381 agcaggagaa cgtagagacc gagcagaagc tcagcgggaa gctgtacctc gtggatctgg
     1441 ccggaagcga gaaggtcagc aagacagggg cagagggagc cgttctggac gaggcaaaga
     1501 atatcaacaa gtcgctgtcg gccctgggga acgtgatctc tgcactggca gagggcacca
     1561 aaagctacgt gccgtaccac gacagcaaaa tgacgaggat tctccaggac tctctggaag
     1621 ggaactgcag gactaccatg ttcatctgct gctcgccgtc cagctacaat gacgcagaga
     1681 ccaagtccac gctcatgttt gggcagcggg cgaagaccat caagaacact gcctcagtga
     1741 atctggagct gactgctgag cagtggaaga agaagtatga gaaggagaag gagaagacca
     1801 agacccagaa ggagacaatt gcgaagctag aggctgagct tagccggtgg cgcaatggag
     1861 agaatgtgcc tgagactgag cgcctggctg gagaggactc agctctggga gctgagctct
     1921 gcgaggagac ccctgtgaat gacaactcat ccattgtggt acgcatcgca cctgaggaaa
     1981 ggcagaaata tgaggaagag atccgccgtc tctacaagca gcttgatgac aaggatgatg
     2041 agatcaacca gcagagccag ctcattgaga agctgaagca gcagatgctg gaccaggaag
     2101 agctgctcgt gtccactcgg ggagacaacg agaaggtcca gcgggagctt agccacctgc
     2161 agtccgagaa cgatgctgcg aaggacgagg tgaaggaagt gctgcaggcc ctagaggagc
     2221 tggcggtcaa ctacgaccag aagtcccagg aggtggagga gaagagccag cagaaccagc
     2281 tgctggtgga cgagctgtcc cagaaagtgg ccaccatgct gtccctggag tccgagctac
     2341 agcggctcca ggaggtcagt ggacaccagc gaaagcggat cgctgaggtg ctgaatgggc
     2401 tgatgaggga cctgagtgag ttcagtgtca tcgtgggcaa cggcgagatt aagctgcccg
     2461 tggagatcag tggggccatc gaggaggagt tcacggtggc ccggctctac atcagcaaga
     2521 tcaagtcgga ggtgaagtcc gtggttaagc gatgtcggca gctggagaac ctccaggtgg
     2581 agtgtcatcg caagatggag gtgaccggta gggagctgtc atcttgccaa ctgctcatct
     2641 cacagcatga ggccaagatc cgttcactca cggagtacat gcagactgtg gagttgaaga
     2701 aacggcacct ggaagagtcc tacgactccc tgagcgatga gcttgccagg ctccaggcgc
     2761 acgaaactgt acacgaggta gctctgaaag acaaggagcc agacacacag gacgcggagg
     2821 aggtgaagaa ggccctggaa ctacagatgg agaatcaccg tgaggcccat caccggcagc
     2881 tggcccgcct ccgggatgag attaatgaga aacagaaaac cattgatgag ctgaaagacc
     2941 tgaaccagaa gctccagtta gagctggaga agcttcaggc cgactatgag aggctgaaga
     3001 atgaagagaa tgagaagagc gccaagctcc aggagctgac atttctgtat gagcgacatg
     3061 agcagtccaa gcaggacctc aaggggctgg aggagacagt tgcccgtgaa ctccagaccc
     3121 tccacaacct tcgcaagctg ttcgttcaag acgtcacgac tcgagtcaag aaaagtgcag
     3181 aaatggagcc cgaggacagt ggggggattc attcccaaaa gcagaagatc tcctttcttg
     3241 agaacaacct ggaacagctt acaaaggttc acaaacagct ggtacgtgac aatgcagatc
     3301 tgcgttgtga gcttcctaaa ttggaaaaac gacttcgggc tacggctgag agagttaagg
     3361 ccctggaggg tgcactgaag gaggccaagg agggtgctat gaaggacaag cgtagatacc
     3421 agcaggaggt ggaccgcatc aaggaagccg tgcggtacaa gagctccggc aagcggggcc
     3481 attctgccca gatcgctaag cctgtgaggc ctggccacta tcctgcctcc tcacccacca
     3541 acccctacgg cacccggagc cccgagtgta tcagctacac caacaacctc ctccagaact
     3601 accagaacct gcacctgcag gctgcgccta gctccacttc agatatgtac tttgccagta
     3661 gcggagccac atctgttgcc cccttggctt cctaccagaa ggccaacatg gacaatggaa
     3721 atgccacaga tatcaacgac aacaggagtg acctgccgtg tggctatgag gctgaggacc
     3781 aggccaagct tttccctctc caccaagaga cagcagccag ttagaattct gcagatatcc
     3841 agcacagtgg cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac
     3901 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3961 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     4021 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     4081 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     4141 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     4201 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4261 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4321 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4381 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4441 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4501 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4561 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     4621 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4681 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4741 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4801 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4861 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4921 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4981 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     5041 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     5101 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     5161 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     5221 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5281 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5341 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5401 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5461 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5521 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5581 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5641 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5701 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5761 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5821 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5881 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5941 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     6001 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6061 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     6121 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6181 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6241 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6301 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6361 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6421 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6481 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6541 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6601 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6661 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6721 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6781 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6841 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6901 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6961 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     7021 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7081 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7141 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7201 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7261 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7321 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7381 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7441 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7501 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7561 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7621 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7681 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7741 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7801 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7861 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7921 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7981 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     8041 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8101 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8161 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8221 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8281 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     8341 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     8401 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     8461 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     8521 ggccagatat acgcgtt
//
