LOCUS       sequence-226628-        8544 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 8544)
  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 8544)
  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..8544
                     /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(3109..3682,3683..3685,3686..3694)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3276..3293)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             join(3707..3709,3710..4423)
                     /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(3749..3770)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4010..4029)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4357..4378
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          4468..4600
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4583..4602
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    4609..5197
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4662..4682)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5080..5091)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5224..5241)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5230..5341
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5568..5587)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5633..5688
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5687..5706)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6047..6063)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6060..6082)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6071..6087
                     /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(6095..6101,6102..6119,6120..6125))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6140..6161
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6220..6415
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6266..6401
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6421..6555
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6458..6477)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6512..6531
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6623..6640)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6794..7382)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6874..6893)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7553..8344,8345..8413))
                     /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     8176..8195
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8414..8518)
                     /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 aagagccctt cccagatgtc accttcacag tgaccatgcg
     2461 gagacgcatg ggttattatc tgatccaaat gtatatccca agcttgctta tagtgatttt
     2521 gtcatggatc tccttctgga ttaatatgga cgccgctcca gctagggtcg gactgggcat
     2581 caccacagtg ctgacaatga ctactcagag ctcaggcagc cgagccagct tgcccaaggt
     2641 ttcttacgtg aaggccatcg atatctggat ggctgtctgc cttctgtttg tcttcagcgc
     2701 actgctggaa tacgccgctg tcaattttgt gtctcgacag cataaagagc tgttgcggtt
     2761 cagaagaaaa cgacgccacc acaaagagga tgaggcagga gaaggacgct tcaactttag
     2821 cgcctatggt atgggacctg cttgcctcca ggctaaagac ggaatttccg tgaagggagc
     2881 caacaatagc aacacaacca acccaccccc tgctccatct aagagcccgg aggaaatgcg
     2941 caaactcttt attcagagag cgaaaaagat cgacaaaatc tcccggatcg gattccccat
     3001 ggctttcctg attttcaaca tgttttattg gatcatctac aagattgtgc gaagggagga
     3061 cgtacacaac cagtaagcgg ccgcaattcc ccccgccccc cccccccccc cctctccctc
     3121 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     3181 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     3241 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     3301 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     3361 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     3421 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     3481 gatagttgtg gaaagagtca aatggctctc ctaagcgtat tcaacaaggg gctgaaggat
     3541 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     3601 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     3661 ctttgaaaaa cacgatgata atatggccac aaccatgggg gatccggtga gcaagggcga
     3721 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     3781 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     3841 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     3901 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     3961 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     4021 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     4081 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     4141 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     4201 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     4261 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     4321 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     4381 cgccgccggg atcactctcg gcatggacga gctgtacaag taatctagag tcgaggtacc
     4441 gatgaatagc taaggtcgag gccgcaggta agtatcaagg ttacaagaca ggtttaagga
     4501 gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct
     4561 attggtctta ctgacatcca ctttgccttt ctctccacag gtgtcgacaa tcaacctctg
     4621 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     4681 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     4741 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     4801 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     4861 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     4921 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     4981 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     5041 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     5101 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     5161 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattcgagc tcggtacgat
     5221 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     5281 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     5341 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     5401 gggaggattg ggaagacaat agcaggccag cttttgttcc ctttagggtt aattgcgcgc
     5461 ttggcgctaa tcatggtcat agctgtttcc actcctcagg tgcaggctgc ctatcagaag
     5521 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct
     5581 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     5641 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     5701 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
     5761 tatgccatat gctggctgcc atgaacaaag gtggctataa agaggtcatc agtatatgaa
     5821 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     5881 tttttatatt ttgtttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     5941 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     6001 cttatgaaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc
     6061 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6121 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6181 ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac
     6241 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     6301 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     6361 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     6421 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6481 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6541 tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6601 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6661 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6721 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6781 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6841 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6901 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6961 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     7021 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7081 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7141 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7201 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     7261 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7321 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7381 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7441 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7501 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7561 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7621 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7681 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7741 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7801 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7861 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7921 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7981 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     8041 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8101 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8161 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8221 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8281 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8341 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8401 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8461 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8521 catttccccg aaaagtgcca cctg
//
