LOCUS       sequence-80942-e        5239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dominant-negatively inhibits RFX (regulatory factor X) transcription
            factor. Made of mouse RFX1 DNA binding domain (400-527 aa) and 
            C-terminal myc tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5239)
  AUTHORS   Bae BI, Tietjen I, Atabay KD, Evrony GD, Johnson MB, Asare E, Wang 
            PP, Murayama AY, Im K, Lisgo SN, Overman L, Sestan N, Chang BS, 
            Barkovich AJ, Grant PE, Topcu M, Politsky J, Okano H, Piao X, Walsh 
            CA
  TITLE     Evolutionarily dynamic alternative splicing of GPR56 regulates 
            regional cerebral cortical patterning.
  JOURNAL   Science. 2014 Feb 14;343(6172):764-8. doi: 10.1126/science.1244392.
  PUBMED    24531968
REFERENCE   2  (bases 1 to 5239)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2014-02-14- 14"; volume: "343"; 
            issue: "6172"; pages: "764-8"
COMMENT     Sequence Label: pCAG-DN-RFX
FEATURES             Location/Qualifiers
     source          1..5239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1745..1754
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2141..2170
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2263..2282)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2328..2383
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2382..2401)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2742..2758)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2742..2758)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2755..2777)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2766..2782
                     /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(2790..2796,2797..2814,2815..2820))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2835..2856
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2915..3110
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2961..3096
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3116..3250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3153..3172)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3207..3226
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3318..3335)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3489..4077)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3569..3588)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4248..5039,5040..5108))
                     /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     4871..4890
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5109..5213)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcggtacc gatatcgcgg
     1741 ccgcgccacc atggaaatcc aaggcggcta catgctaggc aatgccagcc agtcttactc
     1801 ccacaccacc cgtgcctcac cagccacagt ccagtggctc ctggataatt acgagaccgc
     1861 tgagggcgtg agcctgccac ggagcaccct ctactgccac tacctgctgc actgccaaga
     1921 gcagaagctg gagccagtta atgccgcctc tttcggcaag ctcatccgct cggtattcat
     1981 gggtctgcgc acgcgccgtc tgggcaccag gggcaactct aagtaccact attacggcct
     2041 gcggatcaaa gccagctcac ccctgctgcg gctgatggag gaccagcagc acatggccat
     2101 gcggggccag cccttctccc agaaacagag gctgaagccc gaacaaaaac tcatctcaga
     2161 agaggatctg taactcgagg gccgcactcc tcaggtgcag gctgcctatc agaaggtggt
     2221 ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa
     2281 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
     2341 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     2401 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
     2461 catatgctgg ctgccatgaa caaaggtggc tataaagagg tcatcagtat atgaaacagc
     2521 cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta
     2581 tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac
     2641 tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat
     2701 gaagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg
     2761 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     2821 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     2881 ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag
     2941 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3001 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3061 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt
     3121 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3181 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     3241 tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3301 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3361 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3421 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3481 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3541 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3601 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3661 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     3721 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3781 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3841 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3901 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     3961 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4021 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4081 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4141 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4201 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4261 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4321 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4381 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4441 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4501 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4561 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4621 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4681 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4741 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4801 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4861 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4921 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4981 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5041 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5101 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5161 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5221 ccccgaaaag tgccacctg
//
