LOCUS       sequence-370683-        7542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gi-coupled hM4D DREADD fused with mCherry under the control of 
            CaMKIIa promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7542)
  AUTHORS   Bryan Roth
  TITLE     Roth lab DREADDs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7542)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-CaMKIIa-hM4D(Gi)-mCherry
FEATURES             Location/Qualifiers
     source          1..7542
                     /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"
     regulatory      1470..1479
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2928..2937
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2934..3644
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3079..3097)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3347..3366
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3669..4257
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3722..3742)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4140..4151)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4289..4765
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4488..4507)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4805..4945
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4805..4934
                     /label=AAV2 ITR
     rep_origin      5020..5475
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5107..5126)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5317..5338
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5492..5511)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5611..5633
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5671..5689)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5757..5861
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5862..5930,5931..6722)
                     /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(6080..6099)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6893..7481
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7382..7401
                     /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 cggatcctct agagtcgacg ccaccatggc caacttcaca cctgtcaatg
     1501 gcagctcggg caatcagtcc gtgcgcctgg tcacgtcatc atcccacaat cgctatgaga
     1561 cggtggaaat ggtcttcatt gccacagtga caggctccct gagcctggtg actgtcgtgg
     1621 gcaacatcct ggtgatgctg tccatcaagg tcaacaggca gctgcagaca gtcaacaact
     1681 acttcctctt cagcctggcg tgtgctgatc tcatcatagg cgccttctcc atgaacctct
     1741 acaccgtgta catcatcaag ggctactggc ccctgggcgc cgtggtctgc gacctgtggc
     1801 tggccctgga ctgcgtggtg agcaacgcct ccgtcatgaa ccttctcatc atcagctttg
     1861 accgctactt ctgcgtcacc aagcctctca cctaccctgc ccggcgcacc accaagatgg
     1921 caggcctcat gattgctgct gcctgggtac tgtccttcgt gctctgggcg cctgccatct
     1981 tgttctggca gtttgtggtg ggtaagcgga cggtgcccga caaccagtgc ttcatccagt
     2041 tcctgtccaa cccagcagtg acctttggca cagccattgc tggcttctac ctgcctgtgg
     2101 tcatcatgac ggtgctgtac atccacatct ccctggccag tcgcagccga gtccacaagc
     2161 accggcccga gggcccgaag gagaagaaag ccaagacgct ggccttcctc aagagcccac
     2221 taatgaagca gagcgtcaag aagcccccgc ccggggaggc cgcccgggag gagctgcgca
     2281 atggcaagct ggaggaggcc cccccgccag cgctgccacc gccaccgcgc cccgtggctg
     2341 ataaggacac ttccaatgag tccagctcag gcagtgccac ccagaacacc aaggaacgcc
     2401 cagccacaga gctgtccacc acagaggcca ccacgcccgc catgcccgcc cctcccctgc
     2461 agccgcgggc cctcaaccca gcctccagat ggtccaagat ccagattgtg acgaagcaga
     2521 caggcaatga gtgtgtgaca gccattgaga ttgtgcctgc cacgccggct ggcatgcgcc
     2581 ctgcggccaa cgtggcccgc aagttcgcca gcatcgctcg caaccaggtg cgcaagaagc
     2641 ggcagatggc ggcccgggag cgcaaagtga cacgaacgat ctttgccatt ctgctggcct
     2701 tcatcctcac ctggacgccc tacaacgtca tggtcctggt gaacaccttc tgccagagct
     2761 gcatccctga cacggtgtgg tccattggct actggctctg ctacgtcaac agcaccatca
     2821 accctgcctg ctatgctctg tgcaacgcca cctttaaaaa gaccttccgg cacctgctgc
     2881 tgtgccagta tcggaacatc ggcactgcca ggcgggatcc accggtcgcc accatggtga
     2941 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     3001 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     3061 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     3121 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     3181 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     3241 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     3301 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     3361 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     3421 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     3481 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     3541 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     3601 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaagaattc gatatcaagc
     3661 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     3721 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     3781 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     3841 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     3901 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccggaact ttcgctttcc
     3961 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     4021 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     4081 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     4141 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     4201 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     4261 gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc
     4321 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     4381 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     4441 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     4501 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     4561 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     4621 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     4681 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     4741 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg
     4801 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     4861 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     4921 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     4981 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     5041 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     5101 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     5161 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5221 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5281 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5341 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     5401 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5461 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5521 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     5581 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     5641 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5701 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5761 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5821 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5881 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5941 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     6001 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     6061 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     6121 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     6181 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6241 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6301 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6361 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6421 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6481 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6541 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6601 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6661 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6721 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6781 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6841 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6901 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6961 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     7021 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7081 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7141 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7201 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7261 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7321 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7381 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7441 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7501 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
