LOCUS       sequence-240882-        7623 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 7623)
  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 7623)
  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 LC IRES GFP
FEATURES             Location/Qualifiers
     source          1..7623
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     39..61
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     123..142
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    join(1674..2247,2248..2250,2251..2259)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1841..1858)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             join(2272..2274,2275..2988)
                     /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(2314..2335)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2575..2594)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2922..2943
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          3033..3165
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3148..3167
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3174..3762
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3227..3247)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3645..3656)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3789..3806)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3795..3906
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4002..4022)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4002..4020)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4133..4152)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4198..4253
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4252..4271)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4614..4630)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4614..4630)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4627..4649)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4638..4654
                     /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(4662..4668,4669..4686,4687..4692))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4707..4728
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4787..4982
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4833..4968
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4988..5122
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5025..5044)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5079..5098
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5190..5207)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5361..5949)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5441..5460)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6120..6911,6912..6980))
                     /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     6743..6762
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6981..7085)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7115..7494
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
       61 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
      121 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattcg gtaccgcggg
      181 cccggcgcgc cagctggata tctccggacc cgggctagcg ccaccatgcg ctgttctcca
      241 ggcggcgtgt ggctcgccct ggctgcttcc cttctgcacg ttagcctgca gggtgagttc
      301 cagcgcaaac tgtataagga gcttgttaag aattataacc ccctggagcg gccggtcgca
      361 aatgattccc agccactgac agtgtacttc agcctctcct tgctgcagat catggacgtg
      421 gatgaaaaga accaggtgct gaccactaat atttggttgc agatgtcctg gaccgatcac
      481 tacttgcagt ggaatgtgag cgaataccca ggtgtaaaga ctgtaagatt ccctgacggc
      541 caaatctgga aaccagatat cctgctgtac aacagcgcag acgaaaggtt tgatgcaaca
      601 tttcacacca acgtgggagt caattcttca ggccactgcc tgtacctgcc ccctggaatc
      661 ttcaagtcct catgctatat cgacgtccgc tggtttccct tcgacgtcca gcactgcaaa
      721 ctcaaattcg ggagctggag ctacggcgga tggagcctgg atctgcaaat gcaggaggct
      781 gacatctctg gttacatccc gaatggggag tgggaccttg tgggaatccc cggtaaaaga
      841 agcgagcgat tttatgaatg ctgcaaggaa cccttccctg acgtaacatt cacagttatc
      901 atcagaagaa ggccattgtt ctacgccgtt agtttgttgc tccccagtat ttttctcatg
      961 gtcgtggaca tcgtgggatt ttgtctccca cctgatagcg gggagagggt ctcctttaag
     1021 attaccttgt tgctcggcta ttctgtattt ctgatcatcg tgtccgatac ccttcctgcc
     1081 acaatcggca ctccgctgat aggagtgtat ttcgtcgtgt gtatggcact cctggtgata
     1141 agtctggcgg aaactatctt cattgtacgg ctggtacata agcaggacct gcaaagaccc
     1201 gtgccagact ggttgcgaca ccttgtgctg gacagaattg catggattct gtgtcttggc
     1261 gagcaaccta tggcccaccg gccacctgca acctttcaag ccaacaagac agacgattgt
     1321 agtgggtctg atctgttgcc tgctatgggg aatcactgct cccatgttgg gggaccacaa
     1381 gatttggaaa agaccccacg ggggcgggga tcaccccttc ctcctccccg agaagcctct
     1441 ctcgctgtcc gggggctgct ccaggaactg tcaagcatcc gacattttct ggagaagcgg
     1501 gacgagatgc gcgaagtcgc tcgcgactgg ctgcgagtgg gctacgtcct tgacaggctg
     1561 ctgtttcgga tctacttgct ggcggtgctg gcttattcca ttactctggt gacactctgg
     1621 tccatatggc actacagtta ggcggccgca attccccccg cccccccccc ccccccctct
     1681 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     1741 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     1801 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     1861 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     1921 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     1981 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     2041 agttggatag ttgtggaaag agtcaaatgg ctctcctaag cgtattcaac aaggggctga
     2101 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
     2161 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     2221 ttttcctttg aaaaacacga tgataatatg gccacaacca tgggggatcc ggtgagcaag
     2281 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2341 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     2401 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2461 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     2521 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2581 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2641 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2701 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     2761 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     2821 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     2881 cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2941 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaatc tagagtcgag
     3001 gtaccgatga atagctaagg tcgaggccgc aggtaagtat caaggttaca agacaggttt
     3061 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
     3121 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc gacaatcaac
     3181 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     3241 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     3301 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     3361 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     3421 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     3481 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     3541 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     3601 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     3661 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     3721 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt cgagctcggt
     3781 acgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3841 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3901 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3961 caagggggag gattgggaag acaatagccc agcttttgtt ccctttagtg agggttaatt
     4021 gcgcgcttgg cgtaatcatg gtcatagctk atttcactcc tcaggtgcag gctgcctatc
     4081 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     4141 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     4201 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     4261 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     4321 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     4381 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     4441 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     4501 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     4561 ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag
     4621 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4681 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4741 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt
     4801 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4861 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4921 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4981 aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5041 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5101 tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc
     5161 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5221 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5281 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5341 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5401 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5461 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5521 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5581 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5641 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5701 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5761 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5821 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5881 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5941 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6001 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6061 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6121 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6181 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6241 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6301 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6361 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6421 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6481 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6541 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6601 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6661 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6721 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6781 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6841 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6901 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6961 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7021 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7081 ccgcgcacat ttccccgaaa agtgccacct ggtcgacatt gattattgac tagttattaa
     7141 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     7201 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     7261 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     7321 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     7381 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     7441 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag
     7501 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg
     7561 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg gggggcgcgc
     7621 gcc
//
