LOCUS       sequence-237563-        7364 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Chemogenetic activator expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7364)
  AUTHORS   Magnus CJ, Lee PH, Bonaventura J, Zemla R, Gomez JL, Ramirez MH, Hu 
            X, Galvan A, Basu J, Michaelides M, Sternson SM
  TITLE     Ultrapotent chemogenetics for research and potential clinical 
            applications.
  JOURNAL   Science. 2019 Mar 14. pii: science.aav5282. doi: 
            10.1126/science.aav5282.
  PUBMED    30872534
REFERENCE   2  (bases 1 to 7364)
  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: "Science. 
            2019 Mar 14. pii: science.aav5282. doi: 10.1126/science.aav5282."
COMMENT     Sequence Label: pCAG PSAM4-5HT3 HC IRES EGFP
FEATURES             Location/Qualifiers
     source          1..7364
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     292..314
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     376..395
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    join(1927..2500,2501..2503,2504..2512)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2094..2111)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             join(2525..2527,2528..3241)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2567..2588)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2828..2847)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3175..3196
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          3286..3418
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3401..3420
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3427..4015
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3480..3500)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3898..3909)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4042..4059)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4048..4159
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4255..4275)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4255..4273)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4294..4310)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4386..4405)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4451..4506
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4505..4524)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4867..4883)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4880..4902)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4891..4907
                     /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(4915..4921,4922..4939,4940..4945))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4960..4981
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5040..5235
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5086..5221
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5241..5375
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5278..5297)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5332..5351
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5443..5460)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5614..6202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5694..5713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6373..7164,7165..7233))
                     /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     6996..7015
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7234..7338)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt
       61 gcgagagggc gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc
      121 gccgccgcac cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa
      181 tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc
      241 ggggctgtcc gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct
      301 tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt
      361 tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat
      421 tcggtaccgc gggcccggcg cgccagctgg atatctccgg acccgggcta gcgccaccat
      481 gcgctgttct ccaggcggcg tgtggctcgc cctggctgct tcccttctgc acgttagcct
      541 gcagggtgag ttccagcgca aactgtataa ggagcttgtt aagaattata accccctgga
      601 gcggccggtc gcaaatgatt cccagccact gacagtgtac ttcagcctct ccttgctgca
      661 gatcatggac gtggatgaaa agaaccaggt gctgaccact aatatttggt tgcagatgtc
      721 ctggaccgat cactacttgc agtggaatgt gagcgaatac ccaggtgtaa agactgtaag
      781 attccctgac ggccaaatct ggaaaccaga tatcctgctg tacaacagcg cagacgaaag
      841 gtttgatgca acatttcaca ccaacgtggg agtcaattct tcaggccact gcctgtacct
      901 gccccctgga atcttcaagt cctcatgcta tatcgacgtc cgctggtttc ccttcgacgt
      961 ccagcactgc aaactcaaat tcgggagctg gagctacggc ggatggagcc tggatctgca
     1021 aatgcaggag gctgacatct ctggttacat cccgaatggg gagtgggacc ttgtgggaat
     1081 ccccggtaaa agaagcgagc gattttatga atgctgcaag gaacccttcc ctgacgtaac
     1141 attcacagtt atcatcagaa gaaggccatt gttctacgcc gttagtttgt tgctccccag
     1201 tatttttctc atggtcgtgg acatcgtggg attttgtctc ccacctgata gcggggagag
     1261 ggtctccttt aagattacct tgttgctcgg ctattctgta tttctgatca tcgtgtccga
     1321 tacccttcct gccacaatcg gcactccgct gataggagtg tatttcgtcg tgtgtatggc
     1381 actcctggtg ataagtctgg cggaaactat cttcattgta cggctggtac ataagcagga
     1441 cctgcaaaga cccgtgccag actggttgcg acaccttgtg ctggacagaa ttgcatggat
     1501 tctgtgtctt ggcgagcaac ctatggccca ccggccacct gcaacctttc aagccaacaa
     1561 gacagacgat tgtagtgggt ctgatctgtt gcctgctatg gggaatcact gctcccatgt
     1621 tgggggacca caagatttgg aaaagacccc acgggggcgg ggatcacccc ttcctcctcc
     1681 ccgagaagcc tctctcgctg tccgggggct gctccaggaa ctgtcaagca tccgacattt
     1741 tctggagaag caggacgaga tggacgaagt cgctgccgac tggctgcgag tgggctacgt
     1801 ccttgacagg ctgctgtttc ggatctactt gctggcggtg ctggcttatt ccattactct
     1861 ggtgacactc tggtccatat ggcactacag ttaggcggcc gcaattcccc ccgccccccc
     1921 cccccccccc tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg
     1981 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag
     2041 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
     2101 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
     2161 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
     2221 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
     2281 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct aagcgtattc
     2341 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     2401 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     2461 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggggga
     2521 tccggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     2581 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     2641 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2701 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     2761 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2821 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2881 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2941 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     3001 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     3061 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     3121 ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     3181 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     3241 atctagagtc gaggtaccga tgaatagcta aggtcgaggc cgcaggtaag tatcaaggtt
     3301 acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg
     3361 cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt
     3421 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     3481 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     3541 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     3601 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     3661 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     3721 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     3781 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     3841 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     3901 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     3961 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga
     4021 attcgagctc ggtacgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     4081 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     4141 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     4201 tggggcagga cagcaagggg gaggattggg aagacaatag cccagctttt gttcccttta
     4261 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tcctcaggtg
     4321 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac
     4381 tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     4441 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     4501 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     4561 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     4621 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     4681 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     4741 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     4801 gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta
     4861 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4921 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4981 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg
     5041 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5101 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5161 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5221 ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca
     5281 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5341 ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg
     5401 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5461 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5521 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5581 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5641 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5701 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5761 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5821 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5881 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5941 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6001 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6061 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6121 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6181 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6241 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6301 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6361 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6421 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6481 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6541 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6601 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6661 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6721 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6781 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6841 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6901 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6961 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7021 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7081 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7141 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7201 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7261 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7321 acaaataggg gttccgcgca catttccccg aaaagtgcca cctg
//
