LOCUS       sequence-225661-        6576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Epilepsy promoter enhanced with Arc elements driving GFP with SV40E 
            polyA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6576)
  AUTHORS   Burke CT, Vitko I, Straub J, Nylund EO, Gawda A, Blair K, Sullivan 
            KA, Ergun L, Ottolini M, Patel MK, Perez-Reyes E
  TITLE     EpiPro, a Novel, Synthetic, Activity-Regulated Promoter That Targets
            Hyperactive Neurons in Epilepsy for Gene Therapy Applications.
  JOURNAL   Int J Mol Sci. 2023 Sep 23;24(19):14467. doi: 10.3390/ijms241914467.
  PUBMED    37833914
REFERENCE   2  (bases 1 to 6576)
  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"; journalName: "Int J Mol 
            Sci."; date: "2023-09-23"; volume: "24(19):14467. doi"; pages: " 
            10.3390/ijms241914467"
COMMENT     Sequence Label: scADGFP
FEATURES             Location/Qualifiers
     source          1..6576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    1029..1058
                     /label=NFAT binding site
                     /bound_moiety="NFAT transcription factor"
                     /note="NFAT binding site from the human interleukin 2 
                     (IL-2) gene (Mattila et al., 1990)"
     protein_bind    1089..1118
                     /label=NFAT binding site
                     /bound_moiety="NFAT transcription factor"
                     /note="NFAT binding site from the human interleukin 2 
                     (IL-2) gene (Mattila et al., 1990)"
     regulatory      1392..1401
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1398..1400,1401..1403,1404..2114)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    2288..2409
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   2423..2563
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2423..2552
                     /label=AAV2 ITR
     primer_bind     complement(2587..2604)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2593..2817
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2863..3291
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2950..2969)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3160..3181
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3300..3320)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3305..3634
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3485..3620
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3547..3566
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3682..3729
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3748..4122
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4252..4373
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4422..4438)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4422..4438)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4435..4457)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4446..4462
                     /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(4470..4476,4477..4494,4495..4500))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4515..4536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4824..5412)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4904..4923)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5583..6374,6375..6443))
                     /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     6206..6225
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6444..6548)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgccatt
      241 aatgcagcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg
      301 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggggtt
      361 atcggacgcg ttaatcgggc gcgccttaat taaaggcggg cgagagcctt cctgcgtggg
      421 gaagctcctt gctgcgtcat ggctcagcta ttctcagtct ctctcctttt atggtgccgg
      481 aagcaagcgg ctgctgctgg agagccttcc tgcgtgggga agctccttgc tgcgtcatgg
      541 ctcagctatt ctcagcctct ctccttttat ggtgccggaa gcaggcaggc tgctgctgga
      601 gagccttcct gcgtggggaa gctccttgct gcgtcatggc tcagctattc tcagtctctc
      661 tccttttatg gtgccggaag caagcggctg ctgctggaga gccttcctgc gtggggaagc
      721 tccttgctgc gtcatggctc agctattctc agcctctctc cttttatggt gccggaagca
      781 ggcaggctgc tgctgggcga ttaaaggcgg gcgggtggtg cagagtctgc atgcgtgagg
      841 agctcctggg cgcgtcacag ccgcgctatt ctcagcgtct ctccttttat ggcgccggaa
      901 gtgagctggg gttgctggca gcctggctgg cactggggag agattgcagg tacttggtgg
      961 aaaactccag cgtgtgattc accctcaggt acttggtgga aaactccagc gtgtgattca
     1021 ccctaattgg aggaaaaact gtttcataca gaaggcgtgg aggaaaattt gtttcataca
     1081 gaaggcgtgg aggaaaaact gtttcataca gaaggcgtaa tgcgatcgcg ctagcatgcg
     1141 tgggcgtgtg agagtgtcta tttcgaggca gaggaggtat catatgacag ctcacgtcaa
     1201 atcccatggc cgtcatactg tgacgtcttt cagacacccc attgacgtca atgggagagg
     1261 gcctgcggct gggccaatga gaaaccgggc tcggcgagcc ggccgcccac gggcctcgct
     1321 ggctgcatat ataagcagag ctcgtttagt gaccacgtgc agccagacca agcttccggt
     1381 caattgcgat cgccaccatg gtgtccaagg gcgaagagct cttcaccgga gtggtgccta
     1441 ttctggtcga actggacgga gacgttaacg gccacaaatt ttctgtctcc ggcgagggag
     1501 aaggcgatgc tacctacggc aaactcaccc tgaagttcat ctgcaccacc ggaaagctcc
     1561 ccgtcccctg gcctacactg gtgacaaccc tcacctacgg agtccagtgc ttcagcagat
     1621 accccgatca tatgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtgc
     1681 aggagaggac catcttcttc aaggacgatg gcaactacaa gaccagggcc gaagtgaagt
     1741 tcgagggcga caccctcgtg aacaggatcg agctgaaagg catcgacttc aaggaggacg
     1801 gcaatattct cggccacaag ctggagtaca actacaactc ccacaacgtc tacatcatgg
     1861 ccgacaaaca gaagaacggc atcaaggtga atttcaaaat caggcacaac atcgaggacg
     1921 gctccgtgca gctcgccgat cactaccagc agaacacccc tatcggagat ggacctgtgc
     1981 tgcttcctga caatcactac ctgtccaccc agagcgctct gagcaaggac cccaacgaga
     2041 agagggacca tatggtgctc ctggagttcg tgaccgcagc cggcatcacc ctgggcatgg
     2101 acgagctgta taagttctgc tacgagaacg aggtctgatg tacaagtaaa gcggcccggg
     2161 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     2221 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2281 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2341 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2401 tgtatcttaa cgcggcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     2461 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     2521 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc agggcatcta gaaacccgct
     2581 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2641 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2701 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2761 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     2821 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2881 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2941 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3001 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3061 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3121 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3181 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3241 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3301 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3361 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3421 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3481 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3541 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3601 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3661 tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta
     3721 taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc
     3781 accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg
     3841 gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc
     3901 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg
     3961 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg
     4021 ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg
     4081 gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc
     4141 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4201 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     4261 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4321 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4381 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4441 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4501 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4561 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4621 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4681 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4741 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4801 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4861 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4921 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4981 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5041 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5101 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5161 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5221 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5281 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5341 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5401 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5461 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5521 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5581 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5641 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5701 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5761 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5821 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5881 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5941 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6001 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6061 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6121 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6181 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6241 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6301 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6361 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6421 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6481 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6541 gttccgcgca catttccccg aaaagtgcca cctgac
//
