LOCUS       sequence-422387-        7165 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression vector for overexpression of Rbfox1 in 
            motor neurons. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7165)
  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 7165)
  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-Rbfox1
FEATURES             Location/Qualifiers
     source          1..7165
                     /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(2659..2679)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(2686..2757)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     primer_bind     complement(2800..2819)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          2827..3843
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(2881..2903)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3844..4120)
                     /label=chicken beta-actin promoter
     enhancer        4122..4501
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4532..4636
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4637..4705,4706..5497)
                     /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(4855..4874)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5668..6256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6157..6176
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6410..6427
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6495..6629
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6519..6538)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6573..6592
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6635..6830)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6649..6784
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    6889..6910
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6925..6930,6931..6948,6949..6955)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6963..6979
                     /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     6987..7003
                     /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 acttaagata tggagcaaaa cggttgtatc cccctcggta
     1501 tatactagcc tgcaatgaag aaagaacgag acccacatca tcagcatggc tcctagtctt
     1561 ggcatcggtc aagggcgcaa aagcattcat ggcaccaacg ccgtaggtgg gggctggagc
     1621 aagtgtgtgg tggtaggggt cggcagcata aactcgtccg taactgtcac tgtaggcagc
     1681 ggcagtggca ggggtgggct gggcatagcg gtacgcagca taaccaccat aaatgtctgc
     1741 accataaaat ccatcctggt aaacaacacc gccataggcc gggattgggg gcgggggcgc
     1801 tgcagccctg aaggtgttgt acacggtgcg accacggcct cgaaggtgag cccctcggta
     1861 tgcagctgca gcagtggcgg ccggatatgg gaagccaggc attgcagaag tatatacaag
     1921 tgaactgggg ccactgtaca tggaagatcc ctcctggttg gcctggcaca acagcaccgt
     1981 gcctgcatag aagtcggggc tgtagaccgc gcccacaact ggatttaatt tccagccatt
     2041 ggtgtagggg ttgacagtct tcttatttgt catcacgcgt gctgtcgcat tattaacctc
     2101 gattttacgg ccctctacca cggtaccgtg caatttctcc ctcgccctgt ccgcatccgc
     2161 actattttcg aaagttacga aaccaaatcc cttggagcct cgctcattaa aaataatttc
     2221 aacatctaat attttaccaa attgaccaaa catttgtcgg aggtctggat cccggaaccg
     2281 gaaagggatg ttggacacat gcagccgctt gggctgggac ttgctttctg tgttttcaga
     2341 aggttgtgtc tggggctggc catcggtcgg ggctgcatca tctgtctgtg tggcggtgcc
     2401 ggagacggtc tgcgcactgg tgtcagcact ctgctccgag tgcgtctgtg taggaggata
     2461 caggtttaat gtgtggtcag ggacagtggt ctggccggtg tactctggcg cgggatgagg
     2521 atgaggggcc gtgtattctg cagggatgcc attctggggt ggtgcgaact gcgctgaggc
     2581 ataaggctga gccattgtgt caggggcggc ggctgcttcc tgattacccc tcagctgctc
     2641 tctttcacaa ttaccggtcg attggaagta gaggttctcg ctagctttat cgtcatcgtc
     2701 tttgtagtcc ttgtcgtcgt cgtccttgta atccttatca tcatcatctt tatagtcgct
     2761 agccatggta ccgaattctt tgccaaaatg atgagacagc acaataacca gcacgttgcc
     2821 caggagctgt aggaaaaaga agaaggcatg aacatggtta gcagaggctc tagagccgcc
     2881 ggtcacacgc cagaagccga accccgccct gccccgtccc ccccgaaggc agccgtcccc
     2941 ccgcggacag ccccgaggct ggagagggag aaggggacgg cggcgcggcg acgcacgaag
     3001 gccctccccg cccatttcct tcctgccggc gccgcaccgc ttcgccccgc gcccgctaga
     3061 gggggtgcgg cggcgcctcc cagatttcgg ctccgcacag atttgggaca aaggaagtcc
     3121 ctgcgccctc tcgcacgatt accataaaag gcaatggctg cggctcgccg cgcctcgaca
     3181 gccgccggcg ctccgggggc cgccgcgccc ctcccccgag ccctccccgg cccgaggcgg
     3241 ccccgccccg cccggcaccc ccacctgccg ccaccccccg cccggcacgg cgagccccgc
     3301 gccacgcccc gtacggagcc ccgcacccga agccgggccg tgctcagcaa ctcggggagg
     3361 ggggtgcagg gggggttgca gcccgaccga cgcgcccaca ccccctgctc acccccccac
     3421 gcacacaccc cgcacgcagc ctttgttccc ctcgcagccc cccccgcacc gcggggcacc
     3481 gcccccggcc gcgctcccct cgcgcacact gcggagcgca caaagccccg cgccgcgccc
     3541 gcagcgctca cagccgccgg gcagcgcgga gccgcacgcg gcgctcccca cgcacacaca
     3601 cacgcacgca ccccccgagc cgctcccccc gcacaaaggg ccctcccgga gcccctcaag
     3661 gctttcacgc agccacagaa aagaaacaag ccgtcattaa accaagcgct aattacagcc
     3721 cggaggagaa gggccgtccc gcccgctcac ctgtgggagt aacgcggtca gtcagagccg
     3781 gggcgggcgg cgcgaggcgg cggcggagcg gggcacgggg cgaaggcagc gcgcagcgac
     3841 tcccgcccgc cgcgcgcttc gctttttata gggccgccgc cgccgccgcc tcgccataaa
     3901 aggaaacttt cggagcgcgc cgctctgatt ggctgccgcc gcacctctcc gcctcgcccc
     3961 gccccgcccc tcgccccgcc ccgccccgcc tggcgcgcgc cccccccccc ccccgccccc
     4021 atcgctgcac aaaataatta aaaaataaat aaatacaaaa ttgggggtgg ggaggggggg
     4081 gagatgggga gagtgaagca gaacgtgggg ctcacctcga ccatggtaat agcgatgact
     4141 aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc
     4201 caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact
     4261 tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa
     4321 gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg
     4381 ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat
     4441 gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt
     4501 cgacccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4561 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4621 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4681 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4741 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4801 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4861 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4921 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4981 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5041 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5101 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5161 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5221 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5281 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5341 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5401 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5461 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5521 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5581 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5641 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5701 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5761 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5821 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5881 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5941 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6001 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6061 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6121 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6181 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6241 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6301 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6361 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     6421 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     6481 tcattaatgc agcggatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     6541 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     6601 taaccattat aagctgcaat aaacaagtta gctttttgca aaagcctagg cctccaaaaa
     6661 agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa
     6721 taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg
     6781 ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc ggatccgctg
     6841 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     6901 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     6961 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
     7021 tgggctgcag gtcgagggat ctccataaga gaagagggac agctatgact gggagtagtc
     7081 aggagaggag gaaaaatctg gctagtaaaa catgtaagga aaattttagg gatgttaaag
     7141 aaaaaaataa cacaaaacaa aatat
//
