LOCUS       sequence-232437-        8156 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 8156)
  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 8156)
  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: AAV CAMKII PSAM4 GlyR IRES EGFP
FEATURES             Location/Qualifiers
     source          1..8156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        149..1437
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     intron          1460..1592
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1575..1594
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(2944..3517,3518..3520,3521..3529)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3111..3128)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             join(3542..3544,3545..4258)
                     /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(3584..3605)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3845..3864)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4192..4213
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4283..4871
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4336..4356)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4754..4765)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4903..5379
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5102..5121)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5419..5559
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5419..5548
                     /label=AAV2 ITR
     rep_origin      5634..6089
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5721..5740)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5931..5952
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6106..6125)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6225..6247
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6285..6303)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6371..6475
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6476..6544,6545..7336)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPKTLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6694..6713)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7507..8095
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7996..8015
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtttaaca ttatggcctt aggtcacttc atctccatgg
      181 ggttcttctt ctgattttct agaaaatgag atgggggtgc agagagcttc ctcagtgacc
      241 tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta ctaatcttaa
      301 attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc attagggcag
      361 atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac cacccgccca
      421 gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca gtcctacctt
      481 gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc ttcatggtcc
      541 ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc ctgctgctca
      601 gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt ctgggtctgt
      661 ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg
      721 agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca attcccagag
      781 gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat gaattagctt
      841 cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc tgtctcttca
      901 gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca tcccagaatg
      961 tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat cccatcccgt
     1021 ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg atcacttgtg
     1081 gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct gggggaacag
     1141 ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag gccagggcaa
     1201 agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt accggggcaa
     1261 cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac gaaggctcgc
     1321 gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag tcaagccggt
     1381 tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg gggcagctct
     1441 agagcggtac cggatccagg taagtatcaa ggttacaaga caggtttaag gagaccaata
     1501 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct
     1561 tactgacatc cactttgcct ttctctccac aggtgatgaa ttcgctagcg ccaccatgcg
     1621 ctgttctcca ggcggcgtgt ggctcgccct ggctgcttcc cttctgcacg ttagcctgca
     1681 gggtgagttc cagcgcaaac tgtataagga gcttgttaag aattataacc ccctggagcg
     1741 gccggtcgca aatgattccc agccactgac agtgtacttc agcctctcct tgctgcagat
     1801 catggacgtg gatgaaaaga accaggtgct gaccactaat atttggttgc agatgtcctg
     1861 gaccgatcac tacttgcagt ggaatgtgag cgaataccca ggtgtaaaga ctgtaagatt
     1921 ccctgacggc caaatctgga aaccagatat cctgctgtac aacagcgcag acgaaaggtt
     1981 tgatgcaaca tttcacacca acgtgggagt caattcttca ggccactgcc tgtacctgcc
     2041 ccctggaatc ttcaagtcct catgctatat cgacgtccgc tggtttccct tcgacgtcca
     2101 gcactgcaaa ctcaaattcg ggagctggag ctacggcgga tggagcctgg atctgcaaat
     2161 gcaggaggct gacatctctg gttacatccc gaatggggag tgggaccttg tgggaatccc
     2221 cggtaaaaga agcgagcgat tttatgaatg ctgcaaagag cccttcccag atgtcacctt
     2281 cacagtgacc atgcggagac gcatgggtta ttatctgatc caaatgtata tcccaagctt
     2341 gcttatagtg attttgtcat ggatctcctt ctggattaat atggacgccg ctccagctag
     2401 ggtcggactg ggcatcacca cagtgctgac aatgactact cagagctcag gcagccgagc
     2461 cagcttgccc aaggtttctt acgtgaaggc catcgatatc tggatggctg tctgccttct
     2521 gtttgtcttc agcgcactgc tggaatacgc cgctgtcaat tttgtgtctc gacagcataa
     2581 agagctgttg cggttcagaa gaaaacgacg ccaccacaaa gaggatgagg caggagaagg
     2641 acgcttcaac tttagcgcct atggtatggg acctgcttgc ctccaggcta aagacggaat
     2701 ttccgtgaag ggagccaaca atagcaacac aaccaaccca ccccctgctc catctaagag
     2761 cccggaggaa atgcgcaaac tctttattca gagagcgaaa aagatcgaca aaatctcccg
     2821 gatcggattc cccatggctt tcctgatttt caacatgttt tattggatca tctacaagat
     2881 tgtgcgaagg gaggacgtac acaaccagta agcggccgca attccccccs ccvccccccc
     2941 ccccccctct ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg
     3001 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
     3061 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     3121 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     3181 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     3241 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     3301 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctaag cgtattcaac
     3361 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg
     3421 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac
     3481 ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca tgggagatcc
     3541 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     3601 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     3661 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     3721 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     3781 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     3841 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     3901 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     3961 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     4021 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     4081 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     4141 cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     4201 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaac
     4261 cggtgtcgac aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     4321 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     4381 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4441 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     4501 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4561 aactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4621 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     4681 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     4741 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4801 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4861 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     4921 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     4981 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     5041 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     5101 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     5161 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     5221 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     5281 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     5341 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     5401 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     5461 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     5521 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     5581 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     5641 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5701 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5761 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5821 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     5881 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     5941 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     6001 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     6061 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     6121 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     6181 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     6241 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     6301 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     6361 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     6421 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     6481 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     6541 tgctcaccca aaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     6601 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     6661 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6721 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6781 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6841 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6901 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6961 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     7021 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     7081 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     7141 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     7201 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     7261 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     7321 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     7381 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7441 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7501 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7561 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7621 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     7681 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7741 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7801 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7861 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7921 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7981 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     8041 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     8101 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
