LOCUS       sequence-216007-        6578 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 6578)
  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 6578)
  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..6578
                     /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    1031..1060
                     /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    1091..1120
                     /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      1394..1403
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1400..1402,1403..1405,1406..2116)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    2290..2411
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   2425..2565
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2425..2554
                     /label=AAV2 ITR
     primer_bind     complement(2589..2606)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2595..2819
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2865..3293
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2952..2971)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3162..3183
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3302..3322)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3307..3636
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3487..3622
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3549..3568
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3684..3731
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3750..4124
                     /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    4254..4375
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4424..4440)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4424..4440)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4437..4459)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4448..4464
                     /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(4472..4478,4479..4496,4497..4502))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4517..4538
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4826..5414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4906..4925)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5585..6376,6377..6445))
                     /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     6208..6227
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6446..6550)
                     /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 ttaatcgggg gcgcgcctta attaaaggcg ggcgagagcc ttcctgcgtg
      421 gggaagctcc ttgctgcgtc atggctcagc tattctcagt ctctctcctt ttatggtgcc
      481 ggaagcaagc ggctgctgct ggagagcctt cctgcgtggg gaagctcctt gctgcgtcat
      541 ggctcagcta ttctcagcct ctctcctttt atggtgccgg aagcaggcag gctgctgctg
      601 gagagccttc ctgcgtgggg aagctccttg ctgcgtcatg gctcagctat tctcagtctc
      661 tctcctttta tggtgccgga agcaagcggc tgctgctgga gagccttcct gcgtggggaa
      721 gctccttgct gcgtcatggc tcagctattc tcagcctctc tccttttatg gtgccggaag
      781 caggcaggct gctgctgggc gattaaaggc gggcgggtgg tgcagagtct gcatgcgtga
      841 ggagctcctg ggcgcgtcac agccgcgcta ttctcagcgt ctctcctttt atggcgccgg
      901 aagtgagctg gggttgctgg cagcctggct ggcactgggg agagattgca ggtacttggt
      961 ggaaaactcc agcgtgtgat tcaccctcag gtacttggtg gaaaactcca gcgtgtgatt
     1021 caccctaatt ggaggaaaaa ctgtttcata cagaaggcgt ggaggaaaat ttgtttcata
     1081 cagaaggcgt ggaggaaaaa ctgtttcata cagaaggcgt aatgcgatcg cgctagcatg
     1141 cgtgggcgtg tgagagtgtc tatttcgagg cagaggaggt atcatatgac agctcacgtc
     1201 aaatcccatg gccgtcatac tgtgacgtct ttcagacacc ccattgacgt caatgggaga
     1261 gggcctgcgg ctgggccaat gagaaaccgg gctcggcgag ccggccgccc acgggcctcg
     1321 ctggctgcat atataagcag agctcgttta gtgaccacgt gcagccagac caagcttccg
     1381 gtcaattgcg atcgccacca tggtgtccaa gggcgaagag ctcttcaccg gagtggtgcc
     1441 tattctggtc gaactggacg gagacgttaa cggccacaaa ttttctgtct ccggcgaggg
     1501 agaaggcgat gctacctacg gcaaactcac cctgaagttc atctgcacca ccggaaagct
     1561 ccccgtcccc tggcctacac tggtgacaac cctcacctac ggagtccagt gcttcagcag
     1621 ataccccgat catatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1681 gcaggagagg accatcttct tcaaggacga tggcaactac aagaccaggg ccgaagtgaa
     1741 gttcgagggc gacaccctcg tgaacaggat cgagctgaaa ggcatcgact tcaaggagga
     1801 cggcaatatt ctcggccaca agctggagta caactacaac tcccacaacg tctacatcat
     1861 ggccgacaaa cagaagaacg gcatcaaggt gaatttcaaa atcaggcaca acatcgagga
     1921 cggctccgtg cagctcgccg atcactacca gcagaacacc cctatcggag atggacctgt
     1981 gctgcttcct gacaatcact acctgtccac ccagagcgct ctgagcaagg accccaacga
     2041 gaagagggac catatggtgc tcctggagtt cgtgaccgca gccggcatca ccctgggcat
     2101 ggacgagctg tataagttct gctacgagaa cgaggtctga tgtacaagta aagcggcccg
     2161 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
     2221 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     2281 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2341 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2401 aatgtatctt aacgcggcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     2461 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     2521 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcagggcatc tagaaacccg
     2581 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2641 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     2701 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     2761 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     2821 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     2881 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2941 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3001 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3061 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3121 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3181 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3241 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3301 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3361 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3421 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3481 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3541 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     3601 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     3661 attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag
     3721 tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc
     3781 tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc
     3841 gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca
     3901 gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc
     3961 tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg
     4021 ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc
     4081 cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt
     4141 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4201 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4261 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4321 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4381 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     4441 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4501 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4561 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4621 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4681 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4741 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4801 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4861 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4921 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4981 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5041 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5101 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5161 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5221 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5281 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5341 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5401 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5461 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5521 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5581 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5641 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5701 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5761 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5821 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5881 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5941 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6001 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6061 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6121 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6181 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6241 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6301 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6361 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6421 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6481 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6541 gggttccgcg cacatttccc cgaaaagtgc cacctgac
//
