LOCUS       sequence-267513-        7066 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 7066)
  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 7066)
  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 SYN PSAM4 GlyR IRES EGFP
FEATURES             Location/Qualifiers
     source          1..7066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(427..1015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(507..526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1186..1977,1978..2046))
                     /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     1809..1828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2047..2151)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2177..2632)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2314..2335)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2526..2545
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2704..2833
                     /label=AAV2 ITR
     intron          3426..3558
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3541..3560
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(4916..5489,5490..5492,5493..5501)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5083..5100)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             join(5514..5516,5517..6230)
                     /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(5556..5577)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5817..5836)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6164..6185
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6237..6825
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6290..6310)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6708..6719)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6844..6965
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6855..6874)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6909..6928
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca
       61 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
      121 ccgggcgacc aaagcwctga ggccgcccgg gctttgcccg ggcggcctca gtgagcgagc
      181 gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
      241 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
      301 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
      361 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      421 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      481 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      541 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      601 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      661 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      721 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      781 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      841 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      901 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      961 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1021 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1081 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1141 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1261 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1321 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1381 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1441 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1501 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1621 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1681 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1801 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1861 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2041 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2101 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2161 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     2221 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2281 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2341 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2401 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2461 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     2521 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     2581 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     2641 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     2701 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
     2761 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     2821 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
     2881 tctaggaaga tccgagctcc agtgtgctgg aattcgccct tttcagtatt taaattgacg
     2941 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
     3001 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
     3061 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
     3121 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
     3181 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
     3241 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
     3301 agcggaggag tcgtgtcgtg cctgagagcg cagctgtgct cctgggcacc gcgcagtccg
     3361 cccccgcggc tcctggccag accaccccta ggaccccctg ccccaagtcg cagccgttgg
     3421 atcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt
     3481 cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact
     3541 ttgcctttct ctccacaggt gatgatatca ctagtgctag cgccaccatg cgctgttctc
     3601 caggcggcgt gtggctcgcc ctggctgctt cccttctgca cgttagcctg cagggtgagt
     3661 tccagcgcaa actgtataag gagcttgtta agaattataa ccccctggag cggccggtcg
     3721 caaatgattc ccagccactg acagtgtact tcagcctctc cttgctgcag atcatggacg
     3781 tggatgaaaa gaaccaggtg ctgaccacta atatttggtt gcagatgtcc tggaccgatc
     3841 actacttgca gtggaatgtg agcgaatacc caggtgtaaa gactgtaaga ttccctgacg
     3901 gccaaatctg gaaaccagat atcctgctgt acaacagcgc agacgaaagg tttgatgcaa
     3961 catttcacac caacgtggga gtcaattctt caggccactg cctgtacctg ccccctggaa
     4021 tcttcaagtc ctcatgctat atcgacgtcc gctggtttcc cttcgacgtc cagcactgca
     4081 aactcaaatt cgggagctgg agctacggcg gatggagcct ggatctgcaa atgcaggagg
     4141 ctgacatctc tggttacatc ccgaatgggg agtgggacct tgtgggaatc cccggtaaaa
     4201 gaagcgagcg attttatgaa tgctgcaaag agcccttccc agatgtcacc ttcacagtga
     4261 ccatgcggag acgcatgggt tattatctga tccaaatgta tatcccaagc ttgcttatag
     4321 tgattttgtc atggatctcc ttctggatta atatggacgc cgctccagct agggtcggac
     4381 tgggcatcac cacagtgctg acaatgacta ctcagagctc aggcagccga gccagcttgc
     4441 ccaaggtttc ttacgtgaag gccatcgata tctggatggc tgtctgcctt ctgtttgtct
     4501 tcagcgcact gctggaatac gccgctgtca attttgtgtc tcgacagcat aaagagctgt
     4561 tgcggttcag aagaaaacga cgccaccaca aagaggatga ggcaggagaa ggacgcttca
     4621 actttagcgc ctatggtatg ggacctgctt gcctccaggc taaagacgga atttccgtga
     4681 agggagccaa caatagcaac acaaccaacc caccccctgc tccatctaag agcccggagg
     4741 aaatgcgcaa actctttatt cagagagcga aaaagatcga caaaatctcc cggatcggat
     4801 tccccatggc tttcctgatt ttcaacatgt tttattggat catctacaag attgtgcgaa
     4861 gggaggacgt acacaaccag taagcggccg caattccccc cccccccccc ccccccccct
     4921 ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     4981 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     5041 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     5101 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     5161 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     5221 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     5281 tgagttggat agttgtggaa agagtcaaat ggctctccta agcgtattca acaaggggct
     5341 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     5401 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     5461 ggttttcctt tgaaaaacac gatgataata tggccacaac catgggagat ccggtgagca
     5521 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     5581 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     5641 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     5701 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     5761 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     5821 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     5881 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     5941 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     6001 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     6061 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     6121 cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     6181 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa accggtaatc
     6241 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     6301 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     6361 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     6421 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     6481 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     6541 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     6601 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     6661 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     6721 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     6781 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgccgctt cgagcagaca
     6841 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     6901 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     6961 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg
     7021 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataa
//
