LOCUS       sequence-193611-        7166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV packaging plasmid that expresses HA-tagged hM3D(Gq) 
            excitatory DREADD from the EF1a promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7166)
  AUTHORS   Gomez JL, Bonaventura J, Lesniak W, Mathews WB, Sysa-Shah P, 
            Rodriguez LA, Ellis RJ, Richie CT, Harvey BK, Dannals RF, Pomper MG,
            Bonci A, Michaelides M
  TITLE     Chemogenetics revealed: DREADD occupancy and activation via 
            converted clozapine.
  JOURNAL   Science. 2017 Aug 4;357(6350):503-507. doi: 10.1126/science.aan2475.
  PUBMED    28774929
REFERENCE   2  (bases 1 to 7166)
  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"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2017-08-4- 4"; volume: "357"; issue: 
            "6350"; pages: "503-507"
COMMENT     Sequence Label: pOTTC1328 - pAAV EF1a HA-hM3D(Gq)
FEATURES             Location/Qualifiers
     source          1..7166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   20..149
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        250..1428
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          481..1419
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1376..1396
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1483..1509
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3312..3900
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3365..3385)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3783..3794)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3932..4408
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4131..4150)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4448..4588
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4448..4577
                     /label=AAV2 ITR
     rep_origin      4663..5118
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4750..4769)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4960..4981
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5135..5154)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5254..5276
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5400..5504
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5505..5573,5574..6365)
                     /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     complement(5723..5742)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6536..7124
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7025..7044
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
       61 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      121 agtggccaac tccatcacta ggggttcctg cggccgcacg cgtaagcttt gcaaagatgg
      181 ataaagtttt aaacagagag gaatctttgc agctaatgga ccttctaggt cttgaaagga
      241 gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
      301 aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac
      361 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
      421 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
      481 gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg
      541 ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga
      601 agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt
      661 gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt
      721 ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt
      781 tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt
      841 tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg
      901 ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct
      961 ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg
     1021 tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca
     1081 aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg
     1141 gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg
     1201 cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt
     1261 tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac
     1321 ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag
     1381 cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgagg taccggatcc
     1441 agtcgagaag gtaccggatc ctctagagtc gacgccacca tgtacccata cgatgttcca
     1501 gattacgcta ccttgcacaa taacagtaca acctcgcctt tgtttccaaa catcagctcc
     1561 tcctggatac acagcccctc cgatgcaggg ctgcccccgg gaaccgtcac tcatttcggc
     1621 agctacaatg tttctcgagc agctggcaat ttctcctctc cagacggtac caccgatgac
     1681 cctctgggag gtcataccgt ctggcaagtg gtcttcatcg ctttcttaac gggcatcctg
     1741 gccttggtga ccatcatcgg caacatcctg gtaattgtgt catttaaggt caacaagcag
     1801 ctgaagacgg tcaacaacta cttcctctta agcctggcct gtgccgatct gattatcggg
     1861 gtcatttcaa tgaatctgtt tacgacctac atcatcatga atcgatgggc cttagggaac
     1921 ttggcctgtg acctctggct tgccattgac tgcgtagcca gcaatgcctc tgttatgaat
     1981 cttctggtca tcagctttga cagatacttt tccatcacga ggccgctcac gtaccgagcc
     2041 aaacgaacaa caaagagagc cggtgtgatg atcggtctgg cttgggtcat ctcctttgtc
     2101 ctttgggctc ctgccatctt gttctggcaa tactttgttg gaaagagaac tgtgcctccg
     2161 ggagagtgct tcattcagtt cctcagtgag cccaccatta cttttggcac agccatcgct
     2221 ggtttttata tgcctgtcac cattatgact attttatact ggaggatcta taaggaaact
     2281 gaaaagcgta ccaaagagct tgctggcctg caagcctctg ggacagaggc agagacagaa
     2341 aactttgtcc accccacggg cagttctcga agctgcagca gttacgaact tcaacagcaa
     2401 agcatgaaac gctccaacag gaggaagtat ggccgctgcc acttctggtt cacaaccaag
     2461 agctggaaac ccagctccga gcagatggac caagaccaca gcagcagtga cagttggaac
     2521 aacaatgatg ctgctgcctc cctggagaac tccgcctcct ccgacgagga ggacattggc
     2581 tccgagacga gagccatcta ctccatcgtg ctcaagcttc cgggtcacag caccatcctc
     2641 aactccacca agttaccctc atcggacaac ctgcaggtgc ctgaggagga gctggggatg
     2701 gtggacttgg agaggaaagc cgacaagctg caggcccaga agagcgtgga cgatggaggc
     2761 agttttccaa aaagcttctc caagcttccc atccagctag agtcagccgt ggacacagct
     2821 aagacttctg acgtcaactc ctcagtgggt aagagcacgg ccactctacc tctgtccttc
     2881 aaggaagcca ctctggccaa gaggtttgct ctgaagacca gaagtcagat cactaagcgg
     2941 aaaaggatgt ccctggtcaa ggagaagaaa gcggcccaga ccctcagtgc gatcttgctt
     3001 gccttcatca tcacttggac cccatacaac atcatggttc tggtgaacac cttttgtgac
     3061 agctgcatac ccaaaacctt ttggaatctg ggctactggc tgtgctacat caacagcacc
     3121 gtgaaccccg tgtgctatgc tctgtgcaac aaaacattca gaaccacttt caagatgctg
     3181 ctgctgtgcc agtgtgacaa aaaaaagagg cgcaagcagc agtaccagca gagacagtcg
     3241 gtcatttttc acaagcgcgc acccgagcag gccttgtagg cggccgcgat atcgccatca
     3301 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     3361 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     3421 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     3481 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     3541 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     3601 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     3661 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     3721 cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     3781 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     3841 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     3901 atcgataccg agcgctgctc gagagatcta cgggtggcat ccctgtgacc cctccccagt
     3961 gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt
     4021 aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt
     4081 ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga
     4141 accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca
     4201 agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct
     4261 cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc
     4321 caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc
     4381 gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt gcggaccgag
     4441 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     4501 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     4561 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     4621 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     4681 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     4741 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4801 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4861 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4921 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4981 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     5041 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5101 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     5161 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     5221 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     5281 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     5341 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     5401 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5461 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5521 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5581 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5641 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5701 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5761 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5821 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5881 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5941 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     6001 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     6061 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     6121 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6181 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6241 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6301 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6361 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6421 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6481 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6541 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6601 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6661 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6721 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6781 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6841 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6901 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6961 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7021 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     7081 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7141 cctttttacg gttcctggcc ttttgc
//
