LOCUS       sequence-328025-        7082 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 7082)
  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 7082)
  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: KLC1-Myc
FEATURES             Location/Qualifiers
     source          1..7082
                     /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             2337..2366
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2439..2456)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2445..2669
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2715..3143
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2802..2821)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3012..3033
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3152..3172)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3157..3486
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3337..3472
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3399..3418
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3553..4347
                     /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(3607..3626)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4217..4236
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4521..4642
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4558..4577)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4612..4631
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4691..4707)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4691..4707)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4704..4726)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4715..4731
                     /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(4739..4745,4746..4763,4764..4769))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4784..4805
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4922..4939)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5093..5681)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5173..5192)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5852..6643,6644..6712))
                     /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     6475..6494
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6713..6817)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6892..6911)
                     /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 caccatgtat gacaacatgt
      721 ccaccatggt gtacataaag gaagagaagc tggagaagct cacgcaggat gagatcatct
      781 ctaagaccaa gcaagtgatc caggggctgg aagccctgaa gaatgagcac aactccatcc
      841 tgcagagttt gctggagacg ctgaagtgct tgaagaagga tgacgagagc aacctggtgg
      901 aagagaaatc cagcatgatc cgcaagtccc tggagatgct ggagcttggc ctgagcgagg
      961 cgcaggtgat gatggcgctg tccaatcacc tgaatgctgt ggagtccgag aagcaaaagc
     1021 tccgcgctca ggttcgacgg ctgtgccagg agaaccagtg gctgcgggat gagctggcca
     1081 acacgcagca gaagttgcag aagagcgagc agtcggtggc gcagctggag gaggagaaga
     1141 aacacctgga gttcatgaac cagctgaaga agtacgacga cgacatctcc ccctcggagg
     1201 acaaagactc tgattcttcc aaagagccgt tggatgatct cttcccaaat gacgaggacg
     1261 aaccaggaca aggaatccag cagcagcaca gtagtgctgc ggccgccgcc cagcagggcg
     1321 gctacgagat ccctgcaagg ctgcgcacgc tccacaactt ggtgatccag tatgcttcac
     1381 aggggcgtta cgaggtggcg gtgccactct gcaagcaggc cctggaggat ctggagaaga
     1441 cttccggcca cgaccacccc gatgtggcta ccatgctcaa catcttggcc ctggtgtaca
     1501 gggatcagaa caagtataaa gatgcagcta acctcctgaa cgacgccctg gctatccgcg
     1561 agaaaaccct gggcagagat caccccgcgg tggcagcgac tctgaacaac ctagcagtac
     1621 tgtacggtaa gcgagggaag tacaaggagg cggagccgct gtgtaaacga gccctggaga
     1681 tcagggagaa ggttctggga aaggatcatc ctgatgttgc caaacagtta aataacctgg
     1741 ccctgctgtg ccagaaccag ggcaagtacg aggaggtgga gtattattac cagagggccc
     1801 tgggcatcta ccagacgaag ctggggcccg acgatcccaa cgtggccaag accaagaaca
     1861 acctggcctc ctgttatctg aaacaaggga agttcaagca ggcagaaacg ctgtacaagg
     1921 agattctcac ccgcgcacac gagcgggagt ttggatctgt ggacgacgag aacaagccca
     1981 tctggatgca cgctgaagag agagaggagt gcaaaggcaa gcagaaggac gggtcggctt
     2041 ttggagagta tggcggctgg tataaagcct gcaaagtgga cagtcccacc gtcacaacca
     2101 ccttgaaaaa ccttggagca ctttaccgac ggcaggggaa gtttgaagct gcagagacat
     2161 tggaagaagc cgccatgagg tcacgtaagc agggtcttga caatgttcac aaacagagag
     2221 tggctgaagt gctaaatgac cctgagagca tggagaagcg gaggagccgg gagagtctca
     2281 atatggacgt ggtcaagtac gagagtggcc ctgacggagg ggaggaagcc ggaagcgaac
     2341 aaaaactcat ctcagaagag gatctgtaaa attctgcaga tatccagcac agtggcggcc
     2401 gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     2461 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     2521 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     2581 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     2641 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     2701 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2761 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2821 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2881 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     2941 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3001 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3061 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3121 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3181 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3241 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3301 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3361 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3421 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3481 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     3541 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3601 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3661 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3721 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3781 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3841 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3901 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3961 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4021 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4081 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4141 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4201 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4261 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4321 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4381 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4441 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4501 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     4561 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4621 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     4681 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4741 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4801 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4861 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4921 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4981 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5041 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5101 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5161 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5221 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5281 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5341 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5401 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5461 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5521 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5581 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5641 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5701 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5761 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5821 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5881 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5941 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6001 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6061 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6121 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6181 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6241 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6301 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6361 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6421 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6481 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6541 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6601 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6661 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6721 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6781 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6841 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     6901 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     6961 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     7021 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
     7081 tt
//
