LOCUS       sequence-422406-        6917 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression vector for overexpression of Rbfox3 in 
            motor neurons. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6917)
  AUTHORS   Jacko M, Weyn-Vanhentenryck SM, Smerdon JW, Yan R, Feng H, Williams 
            DJ, Pai J, Xu K, Wichterle H, Zhang C
  TITLE     Rbfox Splicing Factors Promote Neuronal Maturation and Axon Initial 
            Segment Assembly.
  JOURNAL   Neuron. 2018 Feb 21;97(4):853-868.e6. doi: 
            10.1016/j.neuron.2018.01.020. Epub 2018 Feb 1.
  PUBMED    29398366
REFERENCE   2  (bases 1 to 6917)
  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"; doi: 
            "10.1016/j.neuron.2018.01.020"; journalName: "Neuron"; date: 
            "2018-02-21- 21"; volume: "97"; issue: "4"; pages: "853-868.e6"
COMMENT     Sequence Label: pCAGGS-3xFLAG-Rbfox3
FEATURES             Location/Qualifiers
     source          1..6917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     181..200
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    199..254
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     300..319
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     primer_bind     421..437
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     456..476
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        458..476
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    572..683
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     672..689
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    716..1304
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             822..833
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1231..1251
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(1311..1330)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          1313..1445
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             complement(2410..2430)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(2437..2508)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     primer_bind     complement(2551..2570)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          2578..3594
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(2632..2654)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3595..3872)
                     /label=chicken beta-actin promoter
     enhancer        3874..4253
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4284..4388
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4389..4457,4458..5249)
                     /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     complement(4607..4626)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5420..6008
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5909..5928
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6162..6179
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6247..6381
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6271..6290)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6325..6344
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6387..6582)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6401..6536
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    6641..6662
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6677..6682,6683..6700,6701..6707)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6715..6731
                     /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)."
     primer_bind     6739..6755
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aaaaaaaatc taacctcaag tcaaggcttt tctatggaat aaggaatgga cagcaggggg
       61 ctgtttcata tactgatgac ctctttatag ccaacctttg ttcatggcag ccagcatatg
      121 ggcatatgtt gccaaactct aaaccaaata ctcattctga tgttttaaat gatttgccct
      181 cccatatgtc cttccgagtg agagacacaa aaaattccaa cacactattg caatgaaaat
      241 aaatttcctt tattagccag aagtcagatg ctcaaggggc ttcatgatgt ccccataatt
      301 tttggcagag ggaaaaagat ctcagtggta tttgtgagcc agggcattgg ccacaccagc
      361 caccaccttc tgataggcag cctgcacctg aggagtgaat tagcggataa caatttcaca
      421 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
      481 aaaagctggg ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc
      541 cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag
      601 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
      661 tggctggcaa ctagaaggca cagtcgaggc tgatcgtacc gagctcgaat tccaggcggg
      721 gaggcggccc aaagggagat ccgactcgtc tgagggcgaa ggcgaagacg cggaagaggc
      781 cgcagagccg gcagcaggcc gcgggaagga aggtccgctg gattgagggc cgaagggacg
      841 tagcagaagg acgtcccgcg cagaatccag gtggcaacac aggcgagcag ccatggaaag
      901 gacgtcagct tccccgacaa caccacggaa ttgtcagtgc ccaacagccg agcccctgtc
      961 cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caatagggag ggggaaagcg
     1021 aaagtcccgg aaaggagctg acaggtggtg gcaatgcccc aaccagtggg ggttgcgtca
     1081 gcaaacacag tgcacaccac gccacgttgc ctgacaacgg gccacaactc ctcataaaga
     1141 gacagcaacc aggatttata caaggaggag aaaatgaaag ccatacggga agcaatagca
     1201 tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga
     1261 ataccagtca atctttcaca aattttgtaa tccagaggtt gattgtcgac acctgtggag
     1321 agaaaggcaa agtggatgtc agtaagacca ataggtgcct atcagaaacg caagagtctt
     1381 ctctgtctcg acaagcccag tttctattgg tctccttaaa cctgtcttgt aaccttgata
     1441 cttacctgcg gcctcgagtt acttaagtca catggttccg atgctgtagg ttgctgtggg
     1501 gccgatggtg tgatggtaag gatcagcagc ggcatagact ctaccataac tgtcactgta
     1561 ggctgctgcg gtggcggctg ctggctgagc atatctgtaa gctgcatagc ctccataaat
     1621 ctcagcacca taaaatccat cctgatacac gaccgctcca taagttggaa tggggggtgg
     1681 aggtggtgca gctcgaaatg tattatacac agcacggccc cggccccgca ggtgtgcacc
     1741 ccggtaggcg acggcggtgc ccgtggtggg gtaggggaaa ctggtcactg catagaattc
     1801 aggcccatag actgttccta ccacagggtt tagcttccag ccattggcat atgggttccc
     1861 aggcttctta ttggtcatga cccgggctgt ggcattattg acctcaattt tccgtccctc
     1921 tacgatcgtc ccattcagct tctcccgggc tcggtcagca tctgagctag tttcaaaagt
     1981 tacaaaccca aaacccttgg agccccgctc gttaaaaatg atctccacgt ctaaaatttt
     2041 cccgaattgc ccgaacattt gccgcaggtc ggggtccctg aaccggaagg ggatgttgga
     2101 gacatgtagt cgtttgggct gctgcttctc cgtggggtcg gaagggtgga gttgctggtt
     2161 gtctgtctgt gctgcttcat ctgccggcac cgtctgggtc ccagcaatgg gctgtgtgct
     2221 ggcctcagtg cctggctgct caggatgagt ctgtgcgggt gtgtagaggg tcatgccgtg
     2281 ctcgggggga acaggggtct ggccggagta gtcctgggtg ggatgaggcg gaggtggggc
     2341 atattcagct gggattccat tctgaggcgg aggagggtac tgggcagggg ggtagggctg
     2401 ggcaccggtc gattggaagt agaggttctc gctagcttta tcgtcatcgt ctttgtagtc
     2461 cttgtcgtcg tcgtccttgt aatccttatc atcatcatct ttatagtcgc tagccatggt
     2521 accgaattct ttgccaaaat gatgagacag cacaataacc agcacgttgc ccaggagctg
     2581 taggaaaaag aagaaggcat gaacatggtt agcagaggct ctagagccgc cggtcacacg
     2641 ccagaagccg aaccccgccc tgccccgtcc cccccgaagg cagccgtccc cccgcggaca
     2701 gccccgaggc tggagaggga gaaggggacg gcggcgcggc gacgcacgaa ggccctcccc
     2761 gcccatttcc ttcctgccgg cgccgcaccg cttcgccccg cgcccgctag agggggtgcg
     2821 gcggcgcctc ccagatttcg gctccgcaca gatttgggac aaaggaagtc cctgcgccct
     2881 ctcgcacgat taccataaaa ggcaatggct gcggctcgcc gcgcctcgac agccgccggc
     2941 gctccggggg ccgccgcgcc cctcccccga gccctccccg gcccgaggcg gccccgcccc
     3001 gcccggcacc cccacctgcc gccacccccc gcccggcacg gcgagccccg cgccacgccc
     3061 cgtacggagc cccgcacccg aagccgggcc gtgctcagca actcggggag gggggtgcag
     3121 ggggggttgc agcccgaccg acgcgcccac accccctgct caccccccca cgcacacacc
     3181 ccgcacgcag cctttgttcc cctcgcagcc ccccccgcac cgcggggcac cgcccccggc
     3241 cgcgctcccc tcgcgcacac tgcggagcgc acaaagcccc gcgccgcgcc cgcagcgctc
     3301 acagccgccg ggcagcgcgg agccgcacgc ggcgctcccc acgcacacac acacgcacgc
     3361 accccccgag ccgctccccc cgcacaaagg gccctcccgg agcccctcaa ggctttcacg
     3421 cagccacaga aaagaaacaa gccgtcatta aaccaagcgc taattacagc ccggaggaga
     3481 agggccgtcc cgcccgctca cctgtgggag taacgcggtc agtcagagcc ggggcgggcg
     3541 gcgcgaggcg gcggcggagc ggggcacggg gcgaaggcag cgcgcagcga ctcccgcccg
     3601 ccgcgcgctt cgctttttat agggccgccg ccgccgccgc ctcgccataa aaggaaactt
     3661 tcggagcgcg ccgctctgat tggctgccgc cgcacctctc cgcctcgccc cgccccgccc
     3721 ctcgccccgc cccgccccgc ctggcgcgcg cccccccccc ccccccgccc ccatcgctgc
     3781 acaaaataat taaaaaataa ataaatacaa aattgggggt ggggaggggg gggagatggg
     3841 gagagtgaag cagaacgtgg ggctcacctc gaccatggta atagcgatga ctaatacgta
     3901 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     3961 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     4021 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     4081 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     4141 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     4201 aactaatgac cccgtaattg attactatta ataactagtc aataatcaat gtcgacccag
     4261 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4321 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4381 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4441 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4501 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4561 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4621 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4681 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4741 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4801 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4861 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4921 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4981 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5041 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5101 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5161 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5221 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5281 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5341 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5401 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5461 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5521 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5581 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5641 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5701 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5761 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5821 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5881 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5941 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6001 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6061 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6121 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6181 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     6241 gcagcggatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     6301 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     6361 ataagctgca ataaacaagt tagctttttg caaaagccta ggcctccaaa aaagcctcct
     6421 cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa
     6481 attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc
     6541 ggagttaggg gcgggactat ggttgctgac taattgagat gcggatccgc tggcacgaca
     6601 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     6661 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     6721 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cttgggctgc
     6781 aggtcgaggg atctccataa gagaagaggg acagctatga ctgggagtag tcaggagagg
     6841 aggaaaaatc tggctagtaa aacatgtaag gaaaatttta gggatgttaa agaaaaaaat
     6901 aacacaaaac aaaatat
//
