LOCUS       sequence-430610-        8915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ExSYTE (P2A) EGFP with 3'UTR of Rat Arc DTE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8915)
  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 8915)
  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-DTE
FEATURES             Location/Qualifiers
     source          1..8915
                     /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(6087..6111)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    6245..6379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6282..6301)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6336..6355
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6402..6418)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6402..6418)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6415..6437)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6426..6442
                     /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(6450..6456,6457..6474,6475..6480))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(6632..6649)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6803..7391)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6883..6902)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7562..8353,8354..8422))
                     /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     8185..8204
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     8632..8650
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8688..8710)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8810..8829
                     /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 tctagatcgg ctccatgact cagccatgcc aggcggaggg tcccagaggg
     5461 gctgagtcct cagccccagc tgaggcagca gctggagtct tcagagccag gagaatgaca
     5521 ccaggtctca agctgctgag aagtctttcc ggccatgtct ggaaggggta ccaccccagc
     5581 accagcaccg tcccctcctc tcttgaagct gcctgcacag aggttccaag acactttcaa
     5641 ggcagagaaa ataggattac aaagaggagg tgcctggcag agggcagcac ccagctcagc
     5701 ctcagagctg aaggtgaaga caagccagcg tgaaaccccg ggtctgccac gaatgcccgc
     5761 tccgctggcc actcaccagc tgcctgccac aagccactgc agcttgagca gggtctgtgc
     5821 cctctcagca cagagcccag ttcgctgcgt ggcctttggc ccccgccaga accttgcagg
     5881 agccttaagg ttcgggccct agcccagcct gaccttacct gctgtgccct gcctgctggt
     5941 caagtccagt cccaggagac cccatgcctt ggctcctagg ctgttccagg cacttccctg
     6001 acctgccggg tgattgccca gctggaacct catccacacc ccagcaccaa ccacctcgtg
     6061 ttggtaactg ctcgtgtctt ctagacaccc agctttcttg tacaaagtgg tctcgacctg
     6121 caggatatcg aattcattga tcataatcag ccataccaca tttgtagagg ttttacttgc
     6181 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     6241 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6301 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6361 atcttatcat gtctggatcc gtaccgagct cgcgtaatca tgtcatagct gtttcctgtg
     6421 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6481 gcctggggtg cctaatgagt gagctaacta cattaattgc gttgcgctca ctgcccgctt
     6541 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6601 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6661 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6721 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6781 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6841 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6901 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6961 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     7021 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7081 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7141 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7201 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     7261 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7321 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7381 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7441 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7501 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7561 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7621 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7681 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7741 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7801 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7861 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7921 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7981 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8041 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     8101 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8161 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8221 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8281 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8341 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8401 cacggaaatg ttgaatactc atactcttcc tttttcaata agcggccgcg gccatgccgg
     8461 ccactagtct cgagttatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8521 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8581 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     8641 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     8701 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     8761 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     8821 cagattgtac tgagagtgca ccatatgaag acgtcgcctc ctcactactt ctggaatagc
     8881 tcagaggccg aggcggcctc ggcctctgca taaat
//
