LOCUS       sequence-233645-        8664 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chemogenetic activator expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8664)
  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 8664)
  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..8664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    join(3229..3802,3803..3805,3806..3814)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3396..3413)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             join(3827..3829,3830..4543)
                     /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(3869..3890)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4130..4149)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4477..4498
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          4588..4720
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4703..4722
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    4729..5317
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4782..4802)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5200..5211)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5344..5361)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5350..5461
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5688..5707)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5753..5808
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5807..5826)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6167..6183)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6180..6202)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6191..6207
                     /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(6215..6221,6222..6239,6240..6245))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6260..6281
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6340..6535
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6386..6521
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6541..6675
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6578..6597)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6632..6651
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6743..6760)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6914..7502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6994..7013)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7673..8464,8465..8533))
                     /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     8296..8315
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8534..8638)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg ggcgagggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcggtacc gcgggcccgg
     1741 cgcgccagct ggatatctcc ggacccgggc tagcgccacc atgcgctgtt ctccaggcgg
     1801 cgtgtggctc gccctggctg cttcccttct gcacgttagc ctgcagggtg agttccagcg
     1861 caaactgtat aaggagcttg ttaagaatta taaccccctg gagcggccgg tcgcaaatga
     1921 ttcccagcca ctgacagtgt acttcagcct ctccttgctg cagatcatgg acgtggatga
     1981 aaagaaccag gtgctgacca ctaatatttg gttgcagatg tcctggaccg atcactactt
     2041 gcagtggaat gtgagcgaat acccaggtgt aaagactgta agattccctg acggccaaat
     2101 ctggaaacca gatatcctgc tgtacaacag cgcagacgaa aggtttgatg caacatttca
     2161 caccaacgtg ggagtcaatt cttcaggcca ctgcctgtac ctgccccctg gaatcttcaa
     2221 gtcctcatgc tatatcgacg tccgctggtt tcccttcgac gtccagcact gcaaactcaa
     2281 attcgggagc tggagctacg gcggatggag cctggatctg caaatgcagg aggctgacat
     2341 ctctggttac atcccgaatg gggagtggga ccttgtggga atccccggta aaagaagcga
     2401 gcgattttat gaatgctgca aggaaccctt ccctgacgta acattcacag ttatcatcag
     2461 aagaaggcca ttgttctacg ccgttagttt gttgctcccc agtatttttc tcatggtcgt
     2521 ggacatcgtg ggattttgtc tcccacctga tagcggggag agggtctcct ttaagattac
     2581 cttgttgctc ggctattctg tatttctgat catcgtgtcc gatacccttc ctgccacaat
     2641 cggcactccg ctgataggag tgtatttcgt cgtgtgtatg gcactcctgg tgataagtct
     2701 ggcggaaact atcttcattg tacggctggt acataagcag gacctgcaaa gacccgtgcc
     2761 agactggttg cgacaccttg tgctggacag aattgcatgg attctgtgtc ttggcgagca
     2821 acctatggcc caccggccac ctgcaacctt tcaagccaac aagacagacg attgtagtgg
     2881 gtctgatctg ttgcctgcta tggggaatca ctgctcccat gttgggggac cacaagattt
     2941 ggaaaagacc ccacgggggc ggggatcacc ccttcctcct ccccgagaag cctctctcgc
     3001 tgtccggggg ctgctccagg aactgtcaag catccgacat tttctggaga agcaggacga
     3061 gatggacgaa gtcgctgccg actggctgcg agtgggctac gtccttgaca ggctgctgtt
     3121 tcggatctac ttgctggcgg tgctggctta ttccattact ctggtgacac tctggtccat
     3181 atggcactac agttaggcgg ccgcaattcc ccccgccccc cccccccccc cctctccctc
     3241 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     3301 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     3361 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     3421 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     3481 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     3541 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     3601 gatagttgtg gaaagagtca aatggctctc ctaagcgtat tcaacaaggg gctgaaggat
     3661 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     3721 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     3781 ctttgaaaaa cacgatgata atatggccac aaccatgggg gatccggtga gcaagggcga
     3841 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     3901 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     3961 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     4021 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     4081 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     4141 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     4201 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     4261 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     4321 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     4381 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     4441 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     4501 cgccgccggg atcactctcg gcatggacga gctgtacaag taatctagag tcgaggtacc
     4561 gatgaatagc taaggtcgag gccgcaggta agtatcaagg ttacaagaca ggtttaagga
     4621 gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct
     4681 attggtctta ctgacatcca ctttgccttt ctctccacag gtgtcgacaa tcaacctctg
     4741 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     4801 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     4861 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     4921 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     4981 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     5041 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     5101 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     5161 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     5221 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     5281 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattcgagc tcggtacgat
     5341 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     5401 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     5461 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     5521 gggaggattg ggaagacaat agcaggccag cttttgttcc ctttagggtt aattgcgcgc
     5581 ttggcgctaa tcatggtcat agctgtttcc actcctcagg tgcaggctgc ctatcagaag
     5641 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct
     5701 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     5761 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     5821 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
     5881 tatgccatat gctggctgcc atgaacaaag gtggctataa agaggtcatc agtatatgaa
     5941 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     6001 tttttatatt ttgtttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     6061 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     6121 cttatgaaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc
     6181 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6241 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6301 ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac
     6361 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     6421 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     6481 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     6541 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6601 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6661 tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6721 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6781 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6841 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6901 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6961 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7021 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7081 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     7141 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7201 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7261 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7321 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     7381 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7441 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7501 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7561 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7621 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7681 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7741 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7801 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7861 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7921 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7981 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     8041 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     8101 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     8161 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8221 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8281 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8341 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8401 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8461 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8521 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8581 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8641 catttccccg aaaagtgcca cctg
//
