LOCUS       sequence-328019-        7919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged KIF5A protein (1-820aa) in pcDNA3.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7919)
  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 7919)
  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-KIF5A delta Tail
FEATURES             Location/Qualifiers
     source          1..7919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     249..268
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        249..267
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             328..351
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2896..2913)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2902..3126
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3172..3600
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3259..3278)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3469..3490
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3609..3629)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3614..3943
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3794..3929
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3856..3875
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4010..4804
                     /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(4064..4083)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4674..4693
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4978..5099
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5015..5034)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5069..5088
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5148..5164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5148..5164)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5161..5183)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5172..5188
                     /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(5196..5202,5203..5220,5221..5226))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5241..5262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5379..5396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5550..6138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5630..5649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6309..7100,7101..7169))
                     /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     6932..6951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7170..7274)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7349..7368)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7540..7919
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      241 atcgaaatta atacgactca ctatagggag acccaagctg gctagcgttt aaacttaagc
      301 ttggtaccga gctcggatcc caccatggac tacaaagacg atgacgacaa gggatccgga
      361 atggcggaga ctaacaacga atgcagcatc aaggtgcttt gccgatttcg gcccctgaac
      421 caggccgaga ttctgcgggg ggacaagttc atccccattt tccaagggga cgacagcgtc
      481 attattgggg gaaagccata tgtctttgac cgcgtcttcc ccccaaacac cactcaggag
      541 caggtttacc acgcctgtgc catgcagatc gtcaaagacg tccttgctgg ttacaatggc
      601 acaatcttcg cttatggaca gacatcctca gggaaaacgc ataccatgga ggggaagctg
      661 cacgaccctc agctgatggg catcattccc cggatcgctc gagacatctt caaccacatc
      721 tactccatgg atgagaacct tgaattccac attaaggtat cttacttcga gatttacctg
      781 gataagatcc gtgacctttt ggatgtgacc aagacgaacc tgtccgtgca tgaggacaaa
      841 aaccgggtgc cgtttgtcaa gggttgtacc gaacgctttg tgtccagccc agaggagatt
      901 ctggatgtga tcgatgaggg gaagtccaac cgtcacgtag ctgtcaccaa catgaacgag
      961 cacagttctc ggagccacag catcttcctc atcaacatca agcaggagaa cgtagagacc
     1021 gagcagaagc tcagcgggaa gctgtacctc gtggatctgg ccggaagcga gaaggtcagc
     1081 aagacagggg cagagggagc cgttctggac gaggcaaaga atatcaacaa gtcgctgtcg
     1141 gccctgggga acgtgatctc tgcactggca gagggcacca aaagctatgt gccgtaccgc
     1201 gacagcaaaa tgacgaggat tctccaggac tctctggaag ggaactgcag gactaccatg
     1261 ttcatctgct gctcgccgtc cagctacaat gacgcagaga ccaagtccac gctcatgttt
     1321 gggcagcggg cgaagaccat caagaacact gcctcagtga atctggagct gactgctgag
     1381 cagtggaaga agaagtatga gaaggagaag gagaagacca agacccagaa ggagacaatt
     1441 gcgaagctag aggctgagct tagccggtgg cgcaatggag agaatgtgcc tgagactgag
     1501 cgcctggctg gagaggactc agctctggga gctgagctct gcgaggagac ccctgtgaat
     1561 gacaactcat ccattgtggt acgcatcgca cctgaggaaa ggcagaaata tgaggaagag
     1621 atccgccgtc tctacaagca gcttgatgac aaggatgatg agatcaacca gcagagccag
     1681 ctcattgaga agctgaagca gcagatgctg gaccaggaag agctgctcgt gtccactcgg
     1741 ggagacaacg agaaggtcca gcgggagctt agccacctgc agtccgagaa cgatgctgcg
     1801 aaggacgagg tgaaggaagt gctgcaggcc ctagaggagc tggcggtcaa ctacgaccag
     1861 aagtcccagg aggtggagga gaagagccag cagaaccagc tgctggtgga cgagctgtcc
     1921 cagaaagtgg ccaccatgct gtccctggag tccgagctac agcggctcca ggaggtcagt
     1981 ggacaccagc gaaagcggat cgctgaggtg ctgaatgggc tgatgaggga cctgagtgag
     2041 ttcagtgtca tcgtgggcaa cggcgagatt aagctgcccg tggagatcag tggggccatc
     2101 gaggaggagt tcacggtggc ccggctctac atcagcaaga tcaagtcgga ggtgaagtcc
     2161 gtggttaagc gatgtcggca gctggagaac ctccaggtgg agtgtcatcg caagatggag
     2221 gtgaccggta gggagctgtc atcttgccaa ctgctcatct cacagcatga ggccaagatc
     2281 cgttcactca cggagtacat gcagactgtg gagttgaaga aacggcacct ggaagagtcc
     2341 tacgactccc tgagcgatga gcttgccagg ctccaggcgc acgaaactgt acacgaggta
     2401 gctctgaaag acaaggagcc agacacacag gacgcggagg aggtgaagaa ggccctggaa
     2461 ctacagatgg agaatcaccg tgaggcccat caccggcagc tggcccgcct ccgggatgag
     2521 attaatgaga aacagaaaac cattgatgag ctgaaagacc tgaaccagaa gctccagtta
     2581 gagctggaga agcttcaggc cgactatgag aggctgaaga atgaagagaa tgagaagagc
     2641 gccaagctcc aggagctgac atttctgtat gagcgacatg agcagtccaa gcaggacctc
     2701 aaggggctgg aggagacagt tgcccgtgaa ctccagaccc tccacaacct tcgcaagctg
     2761 ttcgttcaag acgtcacgac tcgagtcaag aaaagtgcag aaatggagcc cgaggacagt
     2821 gggtagaatt ctgcagatat ccagcacagt ggcggccgct cgagtctaga gggcccgttt
     2881 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     2941 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3001 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3061 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     3121 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     3181 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3241 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3301 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3361 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3421 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3481 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3541 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3601 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3661 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3721 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3781 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3841 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     3901 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     3961 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4021 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4081 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4141 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     4201 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4261 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4321 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4381 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4441 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4501 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     4561 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4621 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4681 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4741 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4801 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4861 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4921 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     4981 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5041 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5101 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5161 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5221 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5281 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5341 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5401 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5461 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5521 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5581 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5641 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5701 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5761 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5821 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5881 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5941 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6001 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6061 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6121 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6181 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6241 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6301 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6361 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6421 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6481 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6541 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6601 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6661 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6721 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6781 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6841 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6901 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6961 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7021 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7081 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7141 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7201 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7261 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     7321 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     7381 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     7441 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     7501 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
     7561 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     7621 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     7681 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     7741 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     7801 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     7861 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatg
//
