LOCUS       sequence-193612-        6836 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV packaging plasmid that expresses HA-tagged hM4D(Gi) 
            inhibitory DREADD from the EF1a promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6836)
  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 6836)
  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: pOTTC1329 - pAAV EF1a HA-hM4D(Gi)
FEATURES             Location/Qualifiers
     source          1..6836
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   30..159
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        260..1438
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          491..1429
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1386..1406
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1493..1519
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2992..3580
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3045..3065)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3463..3474)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3612..4088
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3811..3830)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4128..4268
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4128..4257
                     /label=AAV2 ITR
     rep_origin      4343..4798
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4430..4449)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4640..4661
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4815..4834)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4934..4956
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4994..5012)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5080..5184
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5185..5253,5254..6045)
                     /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(5403..5422)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6216..6804
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6705..6724
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac
       61 tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
      121 gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgcacg cgtaagcttt
      181 gcaaagatgg ataaagtttt aaacagagag gaatctttgc agctaatgga ccttctaggt
      241 cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac
      301 agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg
      361 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg
      421 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc
      481 cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg
      541 cccttgcgtg ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct
      601 tcgggttgga agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg
      661 tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct
      721 tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc
      781 tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg
      841 tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc
      901 ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg
      961 ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag
     1021 gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc
     1081 agggagctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca
     1141 aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc
     1201 gccgtccagg cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg
     1261 ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc
     1321 agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt
     1381 cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgagg
     1441 taccggatcc agtcgagaag gtaccggatc ctctagagtc gacgccacca tgtacccata
     1501 cgatgttcca gattacgcta tggccaactt cacacctgtc aatggcagct cgggcaatca
     1561 gtccgtgcgc ctggtcacgt catcatccca caatcgctat gagacggtgg aaatggtctt
     1621 cattgccaca gtgacaggct ccctgagcct ggtgactgtc gtgggcaaca tcctggtgat
     1681 gctgtccatc aaggtcaaca ggcagctgca gacagtcaac aactacttcc tcttcagcct
     1741 ggcgtgtgct gatctcatca taggcgcctt ctccatgaac ctctacaccg tgtacatcat
     1801 caagggctac tggcccctgg gcgccgtggt ctgcgacctg tggctggccc tggactgcgt
     1861 ggtgagcaac gcctccgtca tgaaccttct catcatcagc tttgaccgct acttctgcgt
     1921 caccaagcct ctcacctacc ctgcccggcg caccaccaag atggcaggcc tcatgattgc
     1981 tgctgcctgg gtactgtcct tcgtgctctg ggcgcctgcc atcttgttct ggcagtttgt
     2041 ggtgggtaag cggacggtgc ccgacaacca gtgcttcatc cagttcctgt ccaacccagc
     2101 agtgaccttt ggcacagcca ttgctggctt ctacctgcct gtggtcatca tgacggtgct
     2161 gtacatccac atctccctgg ccagtcgcag ccgagtccac aagcaccggc ccgagggccc
     2221 gaaggagaag aaagccaaga cgctggcctt cctcaagagc ccactaatga agcagagcgt
     2281 caagaagccc ccgcccgggg aggccgcccg ggaggagctg cgcaatggca agctggagga
     2341 ggcccccccg ccagcgctgc caccgccacc gcgccccgtg gctgataagg acacttccaa
     2401 tgagtccagc tcaggcagtg ccacccagaa caccaaggaa cgcccagcca cagagctgtc
     2461 caccacagag gccaccacgc ccgccatgcc cgcccctccc ctgcagccgc gggccctcaa
     2521 cccagcctcc agatggtcca agatccagat tgtgacgaag cagacaggca atgagtgtgt
     2581 gacagccatt gagattgtgc ctgccacgcc ggctggcatg cgccctgcgg ccaacgtggc
     2641 ccgcaagttc gccagcatcg ctcgcaacca ggtgcgcaag aagcggcaga tggcggcccg
     2701 ggagcgcaaa gtgacacgaa cgatctttgc cattctgctt gccttcatcc tcacctggac
     2761 gccctacaac gtcatggtcc tggtgaacac cttctgccag agctgcatcc ctgacacggt
     2821 gtggtccatt ggctactggc tctgctacgt caacagcacc atcaaccctg cctgctatgc
     2881 tctgtgcaac gccaccttta aaaagacctt ccggcacctg ctgctgtgcc agtatcggaa
     2941 catcggcact gccaggtagg cggccgcgat atcgccatca agcttatcga taatcaacct
     3001 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     3061 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     3121 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     3181 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     3241 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3301 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     3361 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt
     3421 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3481 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3541 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc
     3601 gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     3661 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     3721 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     3781 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     3841 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
     3901 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     3961 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     4021 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     4081 ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt
     4141 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     4201 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     4261 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4321 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4381 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     4441 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4501 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     4561 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4621 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     4681 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     4741 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     4801 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4861 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4921 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4981 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5041 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5101 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5161 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5221 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5281 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5341 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5401 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5461 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5521 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5581 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5641 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5701 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5761 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5821 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5881 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5941 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6001 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6061 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6121 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6181 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6241 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6301 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6361 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6421 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6481 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6541 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6601 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6661 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6721 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6781 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcct
//
