LOCUS       sequence-459522-        8552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chemogenetic inhibitor expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8552)
  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 8552)
  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 GlyR IRES EGFP
FEATURES             Location/Qualifiers
     source          1..8552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     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    3121..3687
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3138..3689
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3282..3299)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             join(3713..3715,3716..4429)
                     /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(3755..3776)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4016..4035)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4363..4384
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          4474..4606
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4589..4608
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    4615..5203
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4668..4688)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5086..5097)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5230..5247)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5236..5347
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5443..5463)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5443..5461)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5574..5593)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5639..5694
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5693..5712)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6055..6071)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6055..6071)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6068..6090)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6079..6095
                     /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(6103..6109,6110..6127,6128..6133))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6148..6169
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6228..6423
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6274..6409
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6429..6563
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6466..6485)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6520..6539
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6631..6648)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6802..7390)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6882..6901)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7561..8352,8353..8421))
                     /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     8184..8203
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8422..8526)
                     /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 gggsggggcg aggcggagag 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 agggctcggg 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 atgctgcaaa gagcccttcc cagatgtcac cttcacagtg
     2461 accatgcgga gacgcatggg ttattatctg atccaaatgt atatcccaag cttgcttata
     2521 gtgattttgt catggatctc cttctggatt aatatggacg ccgctccagc tagggtcgga
     2581 ctgggcatca ccacagtgct gacaatgact actcagagct caggcagccg agccagcttg
     2641 cccaaggttt cttacgtgaa ggccatcgat atctggatgg ctgtctgcct tctgtttgtc
     2701 ttcagcgcac tgctggaata cgccgctgtc aattttgtgt ctcgacagca taaagagctg
     2761 ttgcggttca gaagaaaacg acgccaccac aaagaggatg aggcaggaga aggacgcttc
     2821 aactttagcg cctatggtat gggacctgct tgcctccagg ctaaagacgg aatttccgtg
     2881 aagggagcca acaatagcaa cacaaccaac ccaccccctg ctccatctaa gagcccggag
     2941 gaaatgcgca aactctttat tcagagagcg aaaaagatcg acaaaatctc ccggatcgga
     3001 ttccccatgg ctttcctgat tttcaacatg ttttattgga tcatctacaa gattgtgcga
     3061 agggaggacg tacacaacca gtaagcggcc gcaattcccc ccgccccccc ccccccnntc
     3121 tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     3181 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     3241 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     3301 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     3361 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     3421 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     3481 gagttggata gttgtggaaa gagtcaaatg gctctcctaa gcgtattcaa caaggggctg
     3541 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
     3601 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     3661 gttttccttt gaaaaacacg atgataatat ggccacaacc atgggggatc cggtgagcaa
     3721 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     3781 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     3841 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     3901 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     3961 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     4021 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     4081 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     4141 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     4201 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     4261 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     4321 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     4381 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat ctagagtcga
     4441 ggtaccgatg aatagctaag gtcgaggccg caggtaagta tcaaggttac aagacaggtt
     4501 taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag
     4561 gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt cgacaatcaa
     4621 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4681 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4741 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4801 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4861 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4921 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4981 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     5041 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     5101 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     5161 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat tcgagctcgg
     5221 tacgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     5281 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     5341 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     5401 gcaaggggga ggattgggaa gacaatagcc cagcttttgt tccctttagt gagggttaat
     5461 tgcgcgcttg kcgkwrwmat kgtyatmssy kwwtycwstc ctcaggtgca ggctgcctat
     5521 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
     5581 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     5641 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     5701 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     5761 gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag
     5821 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
     5881 tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc
     5941 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
     6001 cctcttctct tatggagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata
     6061 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6121 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6181 ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag
     6241 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     6301 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     6361 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     6421 aaaaagctaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6481 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6541 atgtatctta tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg
     6601 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6661 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6721 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6781 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6841 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6901 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6961 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     7021 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7081 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7141 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7201 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     7261 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7321 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7381 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7441 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7501 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7561 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7621 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7681 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7741 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7801 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7861 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7921 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7981 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8041 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8101 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8161 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8221 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8281 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8341 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8401 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8461 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8521 tccgcgcaca tttccccgaa aagtgccacc tg
//
