LOCUS       sequence-233818-        5520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes human full-length REST/NRSF C402S to be expressed in a 
            baculovirus/insect cell expression system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5520)
  AUTHORS   Inui K, Zhao Z, Yuan J, Jayaprakash S, Le LTM, Drakulic S, Sander B,
            Golas MM
  TITLE     Stepwise assembly of functional C-terminal REST/NRSF transcriptional
            repressor complexes as a drug target.
  JOURNAL   Protein Sci. 2017 May;26(5):997-1011. doi: 10.1002/pro.3142. Epub 
            2017 Mar 12.
  PUBMED    28218430
REFERENCE   2  (bases 1 to 5520)
  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: "10.1002/pro.3142"; 
            journalName: "Protein Sci"; date: "2017-05"; volume: "26"; issue: 
            "5"; pages: "997-1011"
COMMENT     Sequence Label: pIDS-RESTmut full-length
FEATURES             Location/Qualifiers
     source          1..5520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..125
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             2741..2758
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3504..3523)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3548..3596
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    3798..3831
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      3833..4189
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             4654..5445
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 cgatactagt atacggacct ttaattcaac ccaacacaat atattatagt taaataagaa
       61 ttattatcaa atcatttgta tattaattaa aatactatac tgtaaattac attttattta
      121 caatcactcg acgaagactt gatcacccgg gatctcgaga tggccaccca ggtaatgggg
      181 cagtcttctg gaggaggagg gctgtttacc agcagtggca acattggaat ggccctgcct
      241 aacgacatgt atgacttgca tgacctttcc aaagctgaac tggccgcacc tcagcttatt
      301 atgctggcaa atgtggcctt aactggggaa gtaaatggca gctgctgtga ttacctggtc
      361 ggtgaagaaa gacagatggc agaactgatg ccggttgggg ataacaactt ttcagatagt
      421 gaagaaggag aaggacttga agagtctgct gatataaaag gtgaacctca tggactggaa
      481 aacatggaac tgagaagttt ggaactcagc gtcgtagaac ctcagcctgt atttgaggca
      541 tcaggtgctc cagatattta cagttcaaat aaagatcttc cccctgaaac acctggagcg
      601 gaggacaaag gcaagagctc gaagaccaaa ccctttcgct gtaagccatg ccaatatgaa
      661 gcagaatctg aagaacagtt tgtgcatcac atcagagttc acagtgctaa gaaatttttt
      721 gtggaagaga gtgcagagaa gcaggcaaaa gccagggaat ctggctcttc cactgcagaa
      781 gagggagatt tctccaaggg ccccattcgc tgtgaccgct gcggctacaa tactaatcga
      841 tatgatcact atacagcaca cctgaaacac cacaccagag ctggggataa tgagcgagtc
      901 tacaagtgta tcatttgcac atacacaaca gtgagcgagt atcactggag gaaacattta
      961 agaaaccatt ttccaaggaa agtatacaca tgtggaaaat gcaactattt ttcagacaga
     1021 aaaaacaatt atgttcagca tgttagaact catacaggag aacgcccata taaatgtgaa
     1081 ctttgtcctt actcaagttc tcagaagact catctaacta gacatatgcg tactcattca
     1141 ggtgagaagc catttaaatg tgatcagtgc agttatgtgg cctctaatca acatgaagta
     1201 acccgccatg caagacaggt tcacaatggg cctaaacctc ttaattgccc acactgtgat
     1261 tacaaaacag cagatagaag caacttcaaa aaacatgtag agctacatgt gaacccacgg
     1321 cagttcaatt gccctgtatg tgactatgca gcttccaaga agagcaatct acagtatcac
     1381 ttcaaatcta agcatcctac ttgtcctaat aaaacaatgg atgtctcaaa agtgaaacta
     1441 aagaaaacca aaaaacgaga ggctgacttg cctgataata ttaccaatga aaaaacagaa
     1501 atagaacaaa caaaaataaa aggggatgtg gctggaaaga aaaatgaaaa gtccgtcaaa
     1561 gcagagaaaa gagatgtctc aaaagagaaa aagccttcta ataatgtgtc agtgatccag
     1621 gtgactacca gaactcgaaa atcagtaaca gaggtgaaag agatggatgt gcatacagga
     1681 agcaattcag aaaaattcag taaaactaag aaaagcaaaa ggaagctgga agttgacagc
     1741 cattctttac atggtcctgt gaatgatgag gaatcttcaa caaaaaagaa aaagaaggta
     1801 gaaagcaaat ccaaaaataa tagtcaggaa gtgccaaagg gtgacagcaa agtggaggag
     1861 aataaaaagc aaaatacttg catgaaaaaa agtacaaaga agaaaactct gaaaaataaa
     1921 tcaagtaaga aaagcagtaa gcctcctcag aaggaacctg ttgagaaggg atctgctcag
     1981 atggaccctc ctcagatggg gcctgctccc acagaggcgg ttcagaaggg gcccattcag
     2041 gtggagccgc cacctcccat ggagcatgct cagatggagg gtgcccagat acggcctgct
     2101 cctgacgagc ctgttcagat ggaggtggtt caggaggggc ctgctcagaa ggagctgctg
     2161 cctcccgtgg agcctgctca gatggtgggt gcccaaattg tacttgctca catggagctg
     2221 cctcctccca tggagactgc tcagacggag gttgcccaaa tggggcctgc tcccatggaa
     2281 cctgctcaga tggaggttgc ccaggtagaa tctgctccca tgcaggtggt ccagaaggag
     2341 cctgttcaga tggagctgtc tcctcccatg gaggtggtcc agaaggagcc tgttcagata
     2401 gagctgtctc ctcccatgga ggtggtccag aaggaacctg ttaagataga gctgtctcct
     2461 cccatagagg tggtccagaa ggagcctgtt cagatggagt tgtctcctcc catgggggtg
     2521 gttcagaagg agcctgctca gagggagcca cctcctccca gagagcctcc ccttcacatg
     2581 gagccaattt ccaaaaagcc tcctctccga aaagataaaa aggaaaagtc taacatgcag
     2641 agtgaaaggg cacggaagga gcaagtcctt attgaagttg gcttagtgcc tgttaaagat
     2701 agctggcttc taaaggaaag tgtaagcaca gaggatctct caccaccatc accaccactg
     2761 ccaaaggaaa atttaagaga agaggcatca ggagaccaaa aattactcaa cacaggtgaa
     2821 ggaaataaag aagcccctct tcagaaagta ggagcagaag aggcagatga gagcctacct
     2881 ggtcttgctg ctaatatcaa cgaatctacc catatttcat cctctggaca aaacttgaat
     2941 acgccagagg gtgaaacttt aaatggtaaa catcagactg acagtatagt ttgtgaaatg
     3001 aaaatggaca ctgatcagaa cacaagagag aatctcactg gtataaattc aacagttgaa
     3061 gaaccagttt caccaatgct tcccccttca gcagtagaag aacgtgaagc agtgtccaaa
     3121 actgcactgg catcacctcc tgctacaatg gcagcaaatg agtctcagga aattgatgaa
     3181 gatgaaggca tccacagcca tgaaggaagt gacctaagtg acaacatgtc agagggtagt
     3241 gatgattctg gattgcatgg ggctcggcca gttccacaag aatctagcag aaaaaatgca
     3301 aaggaagcgt tggcagtcaa agcggctaag ggagattttg tttgtatctt ctgtgatcgt
     3361 tctttcagaa agggaaaaga ttacagcaaa cacctcaatc gccatttggt taatgtgtac
     3421 tatcttgaag aagcagctca agggcaggag taagctagca gctgatgcat agcatgcggt
     3481 accgggagat gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     3541 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     3601 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattggga
     3661 ccaatacgcc cgcgtttctt ccttttcccc accccaaccc ccaagttcgg gtgaaggccc
     3721 agggctcgca gccaacgtcg gggcggcaag ccctgccata gccactacgg gtacgtttaa
     3781 acccatgtgc ctggcagata acttcgtata atgtatgcta tacgaagtta tggtacgtac
     3841 taagctctca tgtttcacgt actaagctct catgtttaac gtactaagct ctcatgttta
     3901 acgaactaaa ccctcatggc taacgtacta agctctcatg gctaacgtac taagctctca
     3961 tgtttcacgt actaagctct catgtttgaa caataaaatt aatataaatc agcaacttaa
     4021 atagcctcta aggttttaag ttttataaga aaaaaaagaa tatataaggc ttttaaagct
     4081 tttaaggttt aacggttgtg gacaacaagc cagggatgta acgcactgag aagcccttag
     4141 agcctctcaa agcaattttg agtgacacag gaacacttaa cggctgacat gggaattagc
     4201 ttcacgctgc cgcaagcact cagggcgcaa gggctgctaa aggaagcgga acacgtagaa
     4261 agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctgggagg cagaataaat
     4321 gatcatatcg tcaattatta cctccacggg gagagcctga gcaaactggc ctcaggcatt
     4381 tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaagta gcgtatgcgc
     4441 tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt
     4501 ttcatggctt gttatgactg tttttttggg gtacagtcta tgcctcgggc atccaagcag
     4561 caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag
     4621 cagggcagtc gccctaaaac aaagttaaac atcatgaggg aagcggtgat cgccgaagta
     4681 tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg
     4741 gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat
     4801 ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac
     4861 cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc
     4921 attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt
     4981 ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt
     5041 gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg
     5101 gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc
     5161 ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg
     5221 ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc
     5281 gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag
     5341 gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt
     5401 gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataatgtct aacaattcgt
     5461 tcaagccgac ggatctatgt cgggtgcgga gaaagaggta atgaaatggc acctaggtat
//
