LOCUS       sequence-430594-        8246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ExSYTE (P2A) EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8246)
  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 8246)
  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: pGW1-ExSYTE-P2A-EGFP
FEATURES             Location/Qualifiers
     source          1..8246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     146..168
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     161..177
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        743..1122
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1123..1326
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1277..1297
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1323..1347
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1462..2288
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
     protein_bind    2312..2336
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4634..4690
                     /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(4691..4693,4694..4696,4697..5410)
                     /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(4691..4693,4694..4696,4697..5407)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4736..4757)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4997..5016)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5344..5365
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5418..5442)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    5576..5710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5613..5632)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5667..5686
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5733..5749)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5733..5749)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5746..5768)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5757..5773
                     /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(5781..5787,5788..5805,5806..5811))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5963..5980)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6134..6722)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6214..6233)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6893..7684,7685..7753))
                     /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     7516..7535
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     7963..7981
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8019..8041)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8141..8160
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaaaaaaatt agtcagggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
       61 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
      121 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
      181 cgtgtcaagg acggtgactg cagtgaataa taaaatgtgt gtttgtccga aatacgcgtt
      241 ttgagatttc tgtcgccgac taaattcatg tcgcgcgata gtggtgttta tcgccgatag
      301 agatggcgat attggaaaaa tcgatatttg aaaatatggc atattgaaaa tgtcgccgat
      361 gtgagtttct gtgtaactga tatcgccatt tttccaaaag tgatttttgg gcatacgcga
      421 tatctggcga tagcgcttat atcgtttacg ggggatggcg atagacgact ttggtgactt
      481 gggcgattct gtgtgtcgca aatatcgcag tttcgatata ggtgacagac gatatgaggc
      541 tatatcgccg atagaggcga catcaagctg gcacatggcc aatgcatatc gatctataca
      601 ttgaatcaat attggccatt agccatatta ttcattggtt atatagcata aatcaatatt
      661 ggctattggc cattgcatac gttgtatcca tatcataata tgtacattta tattggctca
      721 tgtccaacat taccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt
      781 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      841 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      901 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      961 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     1021 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     1081 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     1141 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     1201 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     1261 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     1321 agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
     1381 catagaagac accgggaccg atccagcctc cgcggccggg aacggtgcat tggaacgcgg
     1441 attccccgtg ccaagagtga cgtaagtacc gcctatagag tctataggcc cacccccttg
     1501 gcttcttatg catgctatac tgtttttggc ttggggtcta tacacccccg cttcctcatg
     1561 ttataggtga tggtatagct tagcctatag gtgtgggtta ttgaccatta ttgaccactc
     1621 ccctattggt gacgatactt tccattacta atccataaca tggctctttg ccacaactct
     1681 ctttattggc tatatgccaa tacactgtcc ttcagagact gacacggact ctgtattttt
     1741 acaggatggg gtctcattta ttatttacaa attcacatat acaacaccac cgtccccagt
     1801 gcccgcagtt tttattaaac ataacgtggg atctccacgc gaatctcggg tacgtgttcc
     1861 ggacatgggc tcttctccgg tagcggcgga gcttctacat ccgagccctg ctcccatgcc
     1921 tccagcgact catggtcgct cggcagctcc ttgctcctaa cagtggaggc cagacttagg
     1981 cacagcacga tgcccaccac caccagtgtg ccgcacaagg ccgtggcggt agggtatgtg
     2041 tctgaaaatg agctcgggga gcgggcttgc accgctgacg catttggaag acttaaggca
     2101 gcggcagaag aagatgcagg cagctgagtt gttgtgttct gataagagtc agaggtaact
     2161 cccgttgcgg tgctgttaac ggtggagggc agtgtagtct gagcagtact cgttgctgcc
     2221 gcgcgcgcca ccagacataa tagctgacag actaacagac tgttcctttc catgggtctt
     2281 ttctgcagtc accgtccttg acacgaagct tacaagtttg tacaaaaaag caggctccaa
     2341 gcttaaaatg gagtcattga aacgctggaa tgaagagagg gggttgtggt gcgaaaaggg
     2401 cgttcaggta ctactgacca ccataggcgc ctttgcagct tttggcctca tgaccatcgc
     2461 catcagcact gactactggc tctacacaag agctctcatc tgcaacacca ccaacctcac
     2521 agcaggtgat gatggaccac cccatcgtgg gggcagtggc tcctcagaga agaaggaccc
     2581 tgggggcctc acacattcag gcctctggcg gatatgctgc ctggaagggt tgaaaagagg
     2641 tgtctgcgtg aagatcaacc acttcccgga ggacacggac tacgaccacg acagcgcgga
     2701 gtacctgctc cgagtagtcc gggcctccag catctttcct atcctgagcg ccatcctgct
     2761 gctgctcggg ggcgtgtgcg tagctgcctc tcgcgtctac aaatccaaaa ggaacatcat
     2821 cctgggcgca gggatcctgt tcgtggcagc aggcctgagc aacatcatcg gggtgattgt
     2881 gtacatatcg gccaacgccg gcgagcctgg ccccaagagg gacgaggaga agaaaaacca
     2941 ctactcgtac ggctggtcct tctacttcgg cgggctgtcc ttcatcctgg ctgaggtgat
     3001 cggagtactg gccgtcaaca tctacatcga gcgcagccgc gaggcacact gccaatcacg
     3061 ctcggacctg ctcaaggccg gcggcggcgc gggcggcagt ggcgggagcg gcccctcgag
     3121 cgagcgcagc cgcgaggcaa tgtctagact ggacaagagc aaagtcataa actctgctct
     3181 ggaattactc aatgaagtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct
     3241 gggagttgag cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc
     3301 cctggcaatc gagatgctgg acaggcatca tacccacttc tgccccctgg aaggcgagtc
     3361 atggcaagac tttctgcgga acaacgccaa gtcattccgc tgtgctctcc tctcacatcg
     3421 cgacggggct aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga
     3481 aaatcagctc gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct
     3541 gtccgccgtg ggccacttta cactgggctg cgtattggag gatcaggagc atcaagtagc
     3601 aaaagaggaa agagagacac ctaccaccga ttctatgccc ccacttctga gacaagcaat
     3661 tgagctgttc gaccatcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat
     3721 atgtggcctg gagaaacagc taaagtgcga aagcggcagc ggcaaatttt ggaagaggct
     3781 gagaaaattc ctccggaaga gcatgtctag actggacaag agcaaagtca taaactctgc
     3841 tctggaatta ctcaatgaag tcggtatcga aggcctgacg acaaggaaac tcgctcaaaa
     3901 gctgggagtt gagcagccta ccctgtactg gcacgtgaag aacaagcggg ccctgctcga
     3961 tgccctggca atcgagatgc tggacaggca tcatacccac ttctgccccc tggaaggcga
     4021 gtcatggcaa gactttctgc ggaacaacgc caagtcattc cgctgtgctc tcctctcaca
     4081 tcgcgacggg gctaaagtgc atctcggcac ccgcccaaca gagaaacagt acgaaaccct
     4141 ggaaaatcag ctcgcgttcc tgtgtcagca aggcttctcc ctggagaacg cactgtacgc
     4201 tctgtccgcc gtgggccact ttacactggg ctgcgtattg gaggatcagg agcatcaagt
     4261 agcaaaagag gaaagagaga cacctaccac cgattctatg cccccacttc tgagacaagc
     4321 aattgagctg ttcgaccatc agggagccga acctgccttc cttttcggcc tggaactaat
     4381 catatgtggc ctggagaaac agctaaagtg cgaaagcggc agcactagtg cgggttttct
     4441 cacgctgcac aacgccttcc ccaaggaagc ggcgtccggc gtcacggtca cagtcaccgg
     4501 accgcccgct gcacccgcgc ccgcgcccgc gccgcccgct cctgcagcgc ccgcgcccgg
     4561 gaccctgtcc aaagaggccg cggcgtccaa caccaacacg ctcaacagga aaaccacgcc
     4621 cgtgggaagc ggagctacta acttcagcct gctgaagcag gctggagacg tggaggagaa
     4681 ccctggacct atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     4741 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     4801 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     4861 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     4921 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     4981 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     5041 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     5101 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     5161 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     5221 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     5281 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     5341 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     5401 gtacaagtag tctagacacc cagctttctt gtacaaagtg gtctcgacct gcaggatatc
     5461 gaattcattg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     5521 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     5581 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5641 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5701 tgtctggatc cgtaccgagc tcgcgtaatc atgtcatagc tgtttcctgt gtgaaattgt
     5761 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5821 gcctaatgag tgagctaact acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5881 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5941 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6001 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6061 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6121 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6181 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6241 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6301 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6361 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6421 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6481 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6541 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6601 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6661 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6721 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6781 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6841 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6901 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6961 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7021 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7081 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7141 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7201 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7261 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7321 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7381 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7441 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7501 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7561 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7621 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7681 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7741 gttgaatact catactcttc ctttttcaat aagcggccgc ggccatgccg gccactagtc
     7801 tcgagttatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7861 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7921 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     7981 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     8041 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     8101 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     8161 ctgagagtgc accatatgaa gacgtcgcct cctcactact tctggaatag ctcagaggcc
     8221 gaggcggcct cggcctctgc ataaat
//
