LOCUS       sequence-462390-        8674 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 8674)
  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 8674)
  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..8674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    join(3237..3810,3811..3813,3814..3822)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3404..3421)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             join(3835..3837,3838..4551)
                     /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(3877..3898)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4138..4157)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4485..4506
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          4596..4728
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4711..4730
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    4737..5325
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4790..4810)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5208..5219)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5352..5369)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5358..5469
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5565..5585)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5565..5583)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5696..5715)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5761..5816
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5815..5834)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6177..6193)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6177..6193)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6190..6212)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6201..6217
                     /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(6225..6231,6232..6249,6250..6255))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6270..6291
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6350..6545
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6396..6531
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6551..6685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6588..6607)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6642..6661
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6753..6770)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6924..7512)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7004..7023)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7683..8474,8475..8543))
                     /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     8306..8325
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8544..8648)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggsgcgc gccrggsggg gsggggsggg gsgrggggsg
      541 gggsggggsg rggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg rgggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcggtaccgc
     1741 gggcccggcg cgccagctgg atatctccgg acccgggcta gcgccaccat gcgctgttct
     1801 ccaggcggcg tgtggctcgc cctggctgct tcccttctgc acgttagcct gcagggtgag
     1861 ttccagcgca aactgtataa ggagcttgtt aagaattata accccctgga gcggccggtc
     1921 gcaaatgatt cccagccact gacagtgtac ttcagcctct ccttgctgca gatcatggac
     1981 gtggatgaaa agaaccaggt gctgaccact aatatttggt tgcagatgtc ctggaccgat
     2041 cactacttgc agtggaatgt gagcgaatac ccaggtgtaa agactgtaag attccctgac
     2101 ggccaaatct ggaaaccaga tatcctgctg tacaacagcg cagacgaaag gtttgatgca
     2161 acatttcaca ccaacgtggg agtcaattct tcaggccact gcctgtacct gccccctgga
     2221 atcttcaagt cctcatgcta tatcgacgtc cgctggtttc ccttcgacgt ccagcactgc
     2281 aaactcaaat tcgggagctg gagctacggc ggatggagcc tggatctgca aatgcaggag
     2341 gctgacatct ctggttacat cccgaatggg gagtgggacc ttgtgggaat ccccggtaaa
     2401 agaagcgagc gattttatga atgctgcaag gaacccttcc ctgacgtaac attcacagtt
     2461 atcatcagaa gaaggccatt gttctacgcc gttagtttgt tgctccccag tatttttctc
     2521 atggtcgtgg acatcgtggg attttgtctc ccacctgata gcggggagag ggtctccttt
     2581 aagattacct tgttgctcgg ctattctgta tttctgatca tcgtgtccga tacccttcct
     2641 gccacaatcg gcactccgct gataggagtg tatttcgtcg tgtgtatggc actcctggtg
     2701 ataagtctgg cggaaactat cttcattgta cggctggtac ataagcagga cctgcaaaga
     2761 cccgtgccag actggttgcg acaccttgtg ctggacagaa ttgcatggat tctgtgtctt
     2821 ggcgagcaac ctatggccca ccggccacct gcaacctttc aagccaacaa gacagacgat
     2881 tgtagtgggt ctgatctgtt gcctgctatg gggaatcact gctcccatgt tgggggacca
     2941 caagatttgg aaaagacccc acgggggcgg ggatcacccc ttcctcctcc ccgagaagcc
     3001 tctctcgctg tccgggggct gctccaggaa ctgtcaagca tccgacattt tctggagaag
     3061 cgggacgaga tgcgcgaagt cgctcgcgac tggctgcgag tgggctacgt ccttgacagg
     3121 ctgctgtttc ggatctactt gctggcggtg ctggcttatt ccattactct ggtgacactc
     3181 tggtccatat ggcactacag ttaggcggcc gcaattcccc ccgccccccc cccccccccc
     3241 tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     3301 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     3361 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     3421 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     3481 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     3541 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     3601 gtgagttgga tagttgtgga aagagtcaaa tggctctcct aagcgtattc aacaaggggc
     3661 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     3721 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     3781 tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggggga tccggtgagc
     3841 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     3901 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     3961 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     4021 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     4081 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     4141 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     4201 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     4261 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     4321 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     4381 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     4441 acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     4501 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta atctagagtc
     4561 gaggtaccga tgaatagcta aggtcgaggc cgcaggtaag tatcaaggtt acaagacagg
     4621 tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat
     4681 aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt gtcgacaatc
     4741 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     4801 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     4861 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     4921 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     4981 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     5041 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     5101 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     5161 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     5221 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     5281 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga attcgagctc
     5341 ggtacgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     5401 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     5461 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     5521 cagcaagggg gaggattggg aagacaatag cccagctttt gttcccttta gtgagggtta
     5581 attgcgcgct tggcgtaatc atggtcatag ctkatttcac tcctcaggtg caggctgcct
     5641 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt
     5701 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
     5761 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     5821 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
     5881 tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc
     5941 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
     6001 gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt
     6061 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
     6121 tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca
     6181 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     6241 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     6301 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt
     6361 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     6421 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     6481 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     6541 gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6601 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6661 caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga
     6721 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6781 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6841 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6901 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6961 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7021 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7081 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7141 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7201 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7261 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7321 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     7381 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7441 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7501 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7561 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7621 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7681 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7741 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7801 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7861 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7921 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7981 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     8041 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     8101 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8161 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8221 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8281 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8341 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8401 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8461 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8521 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8581 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8641 gttccgcgca catttccccg aaaagtgcca cctg
//
