LOCUS       sequence-422388-        7108 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression vector for overexpression of Rbfox2 in 
            motor neurons. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7108)
  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 7108)
  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-Rbfox2
FEATURES             Location/Qualifiers
     source          1..7108
                     /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(2602..2622)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(2629..2700)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     primer_bind     complement(2743..2762)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          2770..3786
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(2824..2846)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3787..4063)
                     /label=chicken beta-actin promoter
     enhancer        4065..4444
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4475..4579
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4580..4648,4649..5440)
                     /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(4798..4817)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5611..6199
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6100..6119
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6353..6370
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6438..6572
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6462..6481)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6516..6535
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(6578..6773)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6592..6727
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    6832..6853
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6868..6873,6874..6891,6892..6898)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6906..6922
                     /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     6930..6946
                     /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 gtagggggca aatcggctgt agccacctcg
     1501 gtataaactc gccacagcgc caactccata gctggcggca ggggcgaggg catggtaggg
     1561 gtcagctgtg tacaccctgc cgtaaccgtc gctgtaagcg gctgcagcgg ctgcagcagc
     1621 tgtggctgcg gttgcagtag caggctgtgc atatctgtag gctgcatatc caccatagag
     1681 gtcagcaccg taaaatccgt cctggtaaac cacacctgga taggcgggga tggctgttgg
     1741 aggtaccgct cggactgcac catacactgt ccgccctctg cccctcagat gggctcctcg
     1801 gaaagcagct gccgtggtgg ctgcagttgg gtaagggaag ccaggaatga ttagagggat
     1861 gtaagtgttg atgcctcctc ttcctgacaa gggcacagcc gcctcattgc ctagggacac
     1921 atcagcttga aagctggatg ctgcatataa ctcaggacca tacacagctc caactactgg
     1981 gcttaacttc cagccatttg catatggcgt gaccatcttc ttgttggtca tgacccgtgc
     2041 tgttgcatta ttcacctcga ttttacggcc ctctaccacg gtgccgtgca atttctccct
     2101 ggccctgtct gcatcagcac tattctcgaa agttacgaac ccgaatccct tggagccgcg
     2161 ctcattaaag attatttcca catctaggat tttgccaaac tgcccaaaca tctgccggag
     2221 gtcagggtct cggaagcgga agggaatatt agagacatgt agtcgcttgg gcgtagattt
     2281 actctctgaa ttttcactac tttgtgtctg tgactgttgt ccgtctgtct gtgctccacc
     2341 ttctgtcgtg agagatccat tctggttgct gggtgaattg ctactctgtt ctccatgagg
     2401 ctgcgtactc ccgtagaggg tcaggttatg ttcactggtc tggccggcat agtcctgagt
     2461 gtgtggcact ccatattctg tgggaattcc attttgtgga ggtggtggga acgggatggt
     2521 agtaaaaggc tgaaccattg cgtccggagt tgttgttggc tcctggttac cctgagttac
     2581 cattttcttt ttctcaccgg tcgattggaa gtagaggttc tcgctagctt tatcgtcatc
     2641 gtctttgtag tccttgtcgt cgtcgtcctt gtaatcctta tcatcatcat ctttatagtc
     2701 gctagccatg gtaccgaatt ctttgccaaa atgatgagac agcacaataa ccagcacgtt
     2761 gcccaggagc tgtaggaaaa agaagaaggc atgaacatgg ttagcagagg ctctagagcc
     2821 gccggtcaca cgccagaagc cgaaccccgc cctgccccgt cccccccgaa ggcagccgtc
     2881 cccccgcgga cagccccgag gctggagagg gagaagggga cggcggcgcg gcgacgcacg
     2941 aaggccctcc ccgcccattt ccttcctgcc ggcgccgcac cgcttcgccc cgcgcccgct
     3001 agagggggtg cggcggcgcc tcccagattt cggctccgca cagatttggg acaaaggaag
     3061 tccctgcgcc ctctcgcacg attaccataa aaggcaatgg ctgcggctcg ccgcgcctcg
     3121 acagccgccg gcgctccggg ggccgccgcg cccctccccc gagccctccc cggcccgagg
     3181 cggccccgcc ccgcccggca cccccacctg ccgccacccc ccgcccggca cggcgagccc
     3241 cgcgccacgc cccgtacgga gccccgcacc cgaagccggg ccgtgctcag caactcgggg
     3301 aggggggtgc agggggggtt gcagcccgac cgacgcgccc acaccccctg ctcacccccc
     3361 cacgcacaca ccccgcacgc agcctttgtt cccctcgcag ccccccccgc accgcggggc
     3421 accgcccccg gccgcgctcc cctcgcgcac actgcggagc gcacaaagcc ccgcgccgcg
     3481 cccgcagcgc tcacagccgc cgggcagcgc ggagccgcac gcggcgctcc ccacgcacac
     3541 acacacgcac gcaccccccg agccgctccc cccgcacaaa gggccctccc ggagcccctc
     3601 aaggctttca cgcagccaca gaaaagaaac aagccgtcat taaaccaagc gctaattaca
     3661 gcccggagga gaagggccgt cccgcccgct cacctgtggg agtaacgcgg tcagtcagag
     3721 ccggggcggg cggcgcgagg cggcggcgga gcggggcacg gggcgaaggc agcgcgcagc
     3781 gactcccgcc cgccgcgcgc ttcgcttttt atagggccgc cgccgccgcc gcctcgccat
     3841 aaaaggaaac tttcggagcg cgccgctctg attggctgcc gccgcacctc tccgcctcgc
     3901 cccgccccgc ccctcgcccc gccccgcccc gcctggcgcg cgcccccccc cccccccgcc
     3961 cccatcgctg cacaaaataa ttaaaaaata aataaataca aaattggggg tggggagggg
     4021 ggggagatgg ggagagtgaa gcagaacgtg gggctcacct cgaccatggt aatagcgatg
     4081 actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa
     4141 tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac
     4201 acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg
     4261 aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg
     4321 tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata
     4381 tatgggctat gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa
     4441 tgtcgaccca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4501 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4561 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4621 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4681 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4741 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4801 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4861 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4921 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4981 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5041 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5101 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5161 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5221 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5281 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5341 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5401 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5461 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5521 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5581 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5641 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5701 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5761 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5821 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5881 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5941 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6001 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6061 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6121 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6181 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6241 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6301 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     6361 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     6421 gattcattaa tgcagcggat ccagacatga taagatacat tgatgagttt ggacaaacca
     6481 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     6541 ttgtaaccat tataagctgc aataaacaag ttagcttttt gcaaaagcct aggcctccaa
     6601 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
     6661 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
     6721 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcggatccg
     6781 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     6841 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     6901 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     6961 gcttgggctg caggtcgagg gatctccata agagaagagg gacagctatg actgggagta
     7021 gtcaggagag gaggaaaaat ctggctagta aaacatgtaa ggaaaatttt agggatgtta
     7081 aagaaaaaaa taacacaaaa caaaatat
//
