LOCUS       sequence-430591-        8021 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent expressoin of ExSYTE (P2A) EGFP with 3'UTR of Rat Arc 
            DTE under synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8021)
  AUTHORS   Park J, Berthoux C, Hoyos-Ramirez E, Shan L, Morimoto-Tomita M, Wang
            Y, Castillo PE, Tomita S
  TITLE     Chemogenetic regulation of the TARP-lipid interaction mimics LTP and
            reversibly modifies behavior.
  JOURNAL   Cell Rep. 2023 Aug 29;42(8):112826. doi: 
            10.1016/j.celrep.2023.112826. Epub 2023 Jul 19.
  PUBMED    37471228
REFERENCE   2  (bases 1 to 8021)
  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: "Cell Rep.";
            date: "2023-08-29"; volume: "42(8):112826. doi"; pages: " 
            10.1016/j.celrep.2023.112826. Epub 2023 Jul 19"
COMMENT     Sequence Label: pAAV-Syn-FLEX-ExSYTE
FEATURES             Location/Qualifiers
     source          1..8021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     131..150
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     protein_bind    complement(398..431)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(482..515)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    522..546
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2844..2900
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(2901..2903,2904..2906,2907..3620)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2901..2903,2904..2906,2907..3617)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2946..2967)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3207..3226)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3554..3575
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4297..4321)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    4330..4363
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4414..4447
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     promoter        complement(4489..4936)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   4966..5095
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(5157..5745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5237..5256)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5916..6707,6708..6776))
                     /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     6539..6558
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6777..6881)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6949..6967
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7005..7027)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7127..7146
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7163..7618)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7300..7321)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7512..7531
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   7693..7833
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7704..7833
                     /label=AAV2 ITR
     polyA_signal    join(7894..8021,1..349)
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
ORIGIN
        1 aaaaaaacaa aaattagctg agcctggtca tgcatgcctg gaatcccaac aactcgggag
       61 gctgaggcag gagaatcgct tgaacccagg aggcggagat tgcagtgagc caagattgtg
      121 ccactgcact ccagcttggt tcccaataga ccccgcaggc cctacaggtt gtcttcccaa
      181 cttgcccctt gctccatacc acccccctcc accccataat attatagaag gacacctagt
      241 cagacaaaat gatgcaactt aattttatta ggacaaggct ggtgggcact ggagtggcaa
      301 cttccagggc caggagaggc actggggagg ggtcacaggg atgccacccg tagatctctc
      361 gagcagcgct gctcgaggca agcttgatat cgaattcata acttcgtata gcatacatta
      421 tacgaagtta ttctttgcac cattctaaag aataacagtg ataatttctg ggttaaggca
      481 aataacttcg tataggatac tttatacgaa gttatgctag cacaagtttg tacaaaaaag
      541 caggctccaa gcttaaaatg gagtcattga aacgctggaa tgaagagagg gggttgtggt
      601 gcgaaaaggg cgttcaggta ctactgacca ccataggcgc ctttgcagct tttggcctca
      661 tgaccatcgc catcagcact gactactggc tctacacaag agctctcatc tgcaacacca
      721 ccaacctcac agcaggtgat gatggaccac cccatcgtgg gggcagtggc tcctcagaga
      781 agaaggaccc tgggggcctc acacattcag gcctctggcg gatatgctgc ctggaagggt
      841 tgaaaagagg tgtctgcgtg aagatcaacc acttcccgga ggacacggac tacgaccacg
      901 acagcgcgga gtacctgctc cgagtagtcc gggcctccag catctttcct atcctgagcg
      961 ccatcctgct gctgctcggg ggcgtgtgcg tagctgcctc tcgcgtctac aaatccaaaa
     1021 ggaacatcat cctgggcgca gggatcctgt tcgtggcagc aggcctgagc aacatcatcg
     1081 gggtgattgt gtacatatcg gccaacgccg gcgagcctgg ccccaagagg gacgaggaga
     1141 agaaaaacca ctactcgtac ggctggtcct tctacttcgg cgggctgtcc ttcatcctgg
     1201 ctgaggtgat cggagtactg gccgtcaaca tctacatcga gcgcagccgc gaggcacact
     1261 gccaatcacg ctcggacctg ctcaaggccg gcggcggcgc gggcggcagt ggcgggagcg
     1321 gcccctcgag cgagcgcagc cgcgaggcaa tgtctagact ggacaagagc aaagtcataa
     1381 actctgctct ggaattactc aatgaagtcg gtatcgaagg cctgacgaca aggaaactcg
     1441 ctcaaaagct gggagttgag cagcctaccc tgtactggca cgtgaagaac aagcgggccc
     1501 tgctcgatgc cctggcaatc gagatgctgg acaggcatca tacccacttc tgccccctgg
     1561 aaggcgagtc atggcaagac tttctgcgga acaacgccaa gtcattccgc tgtgctctcc
     1621 tctcacatcg cgacggggct aaagtgcatc tcggcacccg cccaacagag aaacagtacg
     1681 aaaccctgga aaatcagctc gcgttcctgt gtcagcaagg cttctccctg gagaacgcac
     1741 tgtacgctct gtccgccgtg ggccacttta cactgggctg cgtattggag gatcaggagc
     1801 atcaagtagc aaaagaggaa agagagacac ctaccaccga ttctatgccc ccacttctga
     1861 gacaagcaat tgagctgttc gaccatcagg gagccgaacc tgccttcctt ttcggcctgg
     1921 aactaatcat atgtggcctg gagaaacagc taaagtgcga aagcggcagc ggcaaatttt
     1981 ggaagaggct gagaaaattc ctccggaaga gcatgtctag actggacaag agcaaagtca
     2041 taaactctgc tctggaatta ctcaatgaag tcggtatcga aggcctgacg acaaggaaac
     2101 tcgctcaaaa gctgggagtt gagcagccta ccctgtactg gcacgtgaag aacaagcggg
     2161 ccctgctcga tgccctggca atcgagatgc tggacaggca tcatacccac ttctgccccc
     2221 tggaaggcga gtcatggcaa gactttctgc ggaacaacgc caagtcattc cgctgtgctc
     2281 tcctctcaca tcgcgacggg gctaaagtgc atctcggcac ccgcccaaca gagaaacagt
     2341 acgaaaccct ggaaaatcag ctcgcgttcc tgtgtcagca aggcttctcc ctggagaacg
     2401 cactgtacgc tctgtccgcc gtgggccact ttacactggg ctgcgtattg gaggatcagg
     2461 agcatcaagt agcaaaagag gaaagagaga cacctaccac cgattctatg cccccacttc
     2521 tgagacaagc aattgagctg ttcgaccatc agggagccga acctgccttc cttttcggcc
     2581 tggaactaat catatgtggc ctggagaaac agctaaagtg cgaaagcggc agcactagtg
     2641 cgggttttct cacgctgcac aacgccttcc ccaaggaagc ggcgtccggc gtcacggtca
     2701 cagtcaccgg accgcccgct gcacccgcgc ccgcgcccgc gccgcccgct cctgcagcgc
     2761 ccgcgcccgg gaccctgtcc aaagaggccg cggcgtccaa caccaacacg ctcaacagga
     2821 aaaccacgcc cgtgggaagc ggagctacta acttcagcct gctgaagcag gctggagacg
     2881 tggaggagaa ccctggacct atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2941 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     3001 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     3061 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     3121 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3181 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     3241 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3301 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     3361 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     3421 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     3481 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     3541 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     3601 tggacgagct gtacaagtag tctagatcgg ctccatgact cagccatgcc aggcggaggg
     3661 tcccagaggg gctgagtcct cagccccagc tgaggcagca gctggagtct tcagagccag
     3721 gagaatgaca ccaggtctca agctgctgag aagtctttcc ggccatgtct ggaaggggta
     3781 ccaccccagc accagcaccg tcccctcctc tcttgaagct gcctgcacag aggttccaag
     3841 acactttcaa ggcagagaaa ataggattac aaagaggagg tgcctggcag agggcagcac
     3901 ccagctcagc ctcagagctg aaggtgaaga caagccagcg tgaaaccccg ggtctgccac
     3961 gaatgcccgc tccgctggcc actcaccagc tgcctgccac aagccactgc agcttgagca
     4021 gggtctgtgc cctctcagca cagagcccag ttcgctgcgt ggcctttggc ccccgccaga
     4081 accttgcagg agccttaagg ttcgggccct agcccagcct gaccttacct gctgtgccct
     4141 gcctgctggt caagtccagt cccaggagac cccatgcctt ggctcctagg ctgttccagg
     4201 cacttccctg acctgccggg tgattgccca gctggaacct catccacacc ccagcaccaa
     4261 ccacctcgtg ttggtaactg ctcgtgtctt ctagacaccc agctttcttg tacaaagtgg
     4321 tggcgcgcca taacttcgta taatgtatgc tatacgaagt tattaagagg tttcatattg
     4381 ctaatagcag ctacaatcca gctaccattc tgcataactt cgtataaagt atcctatacg
     4441 aagttattcc ggagtcgact ctagagatga tccccgggga attctcgact gcgctctcag
     4501 gcacgacacg actcctccgc tgcccaccgc agactgaggc agcgctgagt cgccggcgcc
     4561 gcagcgcaga tggtcgcgcc cgtgcccccc tatctcgcgc ctcgcgtggt gcggtccggc
     4621 tgggccggcg gcggcgcgga cgcgaccaag gtggccggga aggggagttt gcgggggacc
     4681 ggcgagtgac gtcagcgcgc cttcagtgct gaggcggcgg tggcgcgcgc cgccaggcgg
     4741 gggcgaaggc actgtccgcg gtgctgaagc tggcagtgcg cacgcgcctc gccgcatcct
     4801 gtttcccctc cccctctctg ataggggatg cgcaatttgg ggaatggggg ttgggtgctt
     4861 gtccagtggg tcggggtcgg tcgtcaggta ggcaccccca ccccgcctca tcctggtcct
     4921 aaaacccact tgcactcata cgcagggccc tctgcaggcg gccgcaggaa cccctagtga
     4981 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     5041 tcgcccgacg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggacatg
     5101 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5161 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5221 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5281 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5341 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5401 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5461 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5521 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5581 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5641 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5701 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5761 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5821 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5881 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5941 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6001 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6061 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6121 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6181 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6241 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6301 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6361 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6421 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6481 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6541 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6601 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6661 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6721 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6781 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6841 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6901 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     6961 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     7021 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     7081 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     7141 attgtactga gagtgcacca taaaattgta aacgttaata ttttgttaaa attcgcgtta
     7201 aatttttgtt aaatcagctc attttttaac caatagaccg aaatcggcaa aatcccttat
     7261 aaatcaaaag aatagcccga gatagagttg agtgttgttc cagtttggaa caagagtcca
     7321 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     7381 ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg taaagcacta
     7441 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     7501 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg
     7561 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac
     7621 tatggttgct ttgacgtatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     7681 gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa
     7741 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
     7801 agggagtggc caactccatc actaggggtt cctgcgtccg ctcggtgggt tcctgcgtcc
     7861 gctcggtccg cacgtggtta cctacaaaat cagaaggaca gggaagggag cagtggttca
     7921 cgcctgtaat cccagcaatt tgggaggcca aggtgggtag atcacctgag attaggagtt
     7981 ggagaccagc ctggccaata tggtgaaacc ccgtctctac c
//
