LOCUS       sequence-328024-        7577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged SFPQ Y527A protein in pcDNA3.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7577)
  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 7577)
  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-SFPQY527A
FEATURES             Location/Qualifiers
     source          1..7577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1979..1996)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1985..2209
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2255..2683
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2342..2361)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2552..2573
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2692..2712)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2697..3026
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2877..3012
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2939..2958
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3093..3887
                     /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(3147..3166)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3757..3776
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4061..4182
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4098..4117)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4152..4171
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4231..4247)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4231..4247)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4244..4266)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4255..4271
                     /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(4279..4285,4286..4303,4304..4309))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4324..4345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4462..4479)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4633..5221)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4713..4732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5392..6183,6184..6252))
                     /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     6015..6034
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6253..6357)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6432..6451)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6623..7002
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7003..7206
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7157..7177
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7251..7270
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7251..7269
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7330..7353
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 accaccgcag cagccaccgc cgcagcagcc gccaccgcat cagccgccgc cgcatccaca
       61 gccgcatcag cagcagcagc cgccgccacc gccgcaggac tcttccaagc ccgtcgttgc
      121 tcagggaccc ggccccgctc ccggagtagg cagcgcacca ccagcctcca gctcggcccc
      181 gcccgccact ccaccaacct cgggggcccc gccagggtcc gggccaggcc cgactccgac
      241 cccgccgcct gcagtcacct cggcccctcc cggggcgccg ccacccaccc cgccaagcag
      301 cggggtccct accacacctc ctcaggccgg aggcccgccg cctccgcccg cggcagtccc
      361 gggcccgggt ccagggccta agcagggccc aggtccgggt ggtcccaaag gcggcaaaat
      421 gcctggcggg ccgaagccag gtggcggccc gggcctaagt acgcctggcg gccaccccaa
      481 gccgccgcat cgaggcggcg gggagccccg cgggggccgc cagcaccacc cgccctacca
      541 ccagcagcat caccaggggc ccccgcccgg cgggcccggc ggccgcagcg aggagaagat
      601 ctcggactcg gaggggttta aagccaattt gtctctcttg aggaggcctg gagagaaaac
      661 ttacacacag cgatgtcggt tgtttgttgg gaatctacct gctgatatca cggaggatga
      721 gttcaaaaga ctatttgcta aatatggaga accaggagaa gtttttatca acaaaggcaa
      781 aggattcgga tttattaagc ttgaatctag agctttggct gaaattgcca aagccgaact
      841 ggatgataca cccatgagag gtagacagct tcgagttcgc tttgccacac atgctgctgc
      901 cctttctgtt cgtaatcttt caccttatgt ttccaatgaa ctgttggaag aagcctttag
      961 ccaatttggt cctattgaaa gggctgttgt aatagtggat gatcgtggaa gatctacagg
     1021 gaaaggcatt gttgaatttg cttctaagcc agcagcaaga aaggcatttg aacgatgcag
     1081 tgaaggtgtt ttcttactga cgacaactcc tcgtccagtc attgtggaac cacttgaaca
     1141 actagatgat gaagatggtc ttcctgaaaa acttgcccag aagaatccaa tgtatcaaaa
     1201 ggagagagaa acccctcctc gttttgccca gcatggcacg tttgagtacg aatattctca
     1261 gcgatggaag tctttggatg aaatggaaaa acagcaaagg gaacaagttg aaaaaaacat
     1321 gaaagatgca aaagacaaat tggaaagtga aatggaagat gccgctcatg aacatcaggc
     1381 aaatcttttg cgccaagatc tgatgagacg acaggaagaa ttaagacgca tggaagaact
     1441 tcacaatcaa gaaatgcaga aacgtaaaga aatgcaattg aggcaagagg aggaacgacg
     1501 tagaagagag gaagagatga tgattcgtca acgtgagatg gaagaacaaa tgaggcgcca
     1561 aagagaggaa agttacagcc gaatgggcta catggatcca cgggaaagag acatgcgaat
     1621 gggtggcgga ggagcaatga acatgggaga tccctatggt tcaggaggcc agaaatttcc
     1681 acctctagga ggtggtggtg gcataggtta tgaagctaat cctggcgttc caccagcaac
     1741 catgagtggt tccatgatgg gaagtgacat gcgtactgag cgctttgggc agggaggtgc
     1801 ggggcctgtg ggtggacagg gtcctagagg aatggggcct ggaactccag caggatatgg
     1861 tagagggaga gaagagtacg aaggcccaaa caaaaaaccc cgattttaga attctgcaga
     1921 tatccagcac agtggcggcc gctcgagtct agagggcccg tttaaacccg ctgatcagcc
     1981 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2041 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2101 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2161 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     2221 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     2281 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2341 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2401 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2461 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2521 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2581 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2641 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     2701 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2761 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     2821 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     2881 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     2941 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     3001 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     3061 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3121 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3181 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3241 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     3301 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3361 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3421 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3481 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3541 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3601 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3661 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3721 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     3781 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     3841 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     3901 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     3961 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4021 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     4081 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4141 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     4201 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4261 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4321 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4381 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4441 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4501 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4561 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4621 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4681 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4741 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4801 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4861 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4921 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4981 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5041 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5101 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5161 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5221 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5281 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5341 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5401 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5461 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5521 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5581 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5641 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5701 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5761 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5821 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5881 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5941 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6001 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6061 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6121 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6181 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6241 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6301 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6361 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     6421 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     6481 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     6541 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
     6601 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
     6661 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     6721 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     6781 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     6841 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     6901 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     6961 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     7021 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     7081 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     7141 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     7201 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
     7261 cactataggg agacccaagc tggctagcgt ttaaacttaa gcttggtacc gagctcggat
     7321 cccaccatgg actacaaaga cgatgacgac aagggatccg gaatgtctcg ggatcggttc
     7381 cggagtcgtg gcggtggcgg tggtggcttc cacaggcgtg gaggaggcgg cggccgcggc
     7441 ggcctccacg acttccgttc tccgccgccc ggcatgggcc tcaatcagaa tcgcggcccc
     7501 atgggtcctg gcccgggcca gagcggccct aagcctccga tcccgccacc gcctccacac
     7561 caacagcagc aacagcc
//
