LOCUS       sequence-166302-        7252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gq-coupled hM3D DREADD fused with mCherry under the control of GFAP 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7252)
  AUTHORS   Bryan Roth
  TITLE     Roth lab DREADDs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7252)
  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            "Unpublished"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-GFAP-hM3D(Gq)-mCherry
FEATURES             Location/Qualifiers
     source          1..7252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      2638..2647
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2644..3354
                     /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(2789..2807)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3057..3076
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3379..3967
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3432..3452)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3850..3861)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3999..4475
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4198..4217)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4515..4655
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4515..4644
                     /label=AAV2 ITR
     rep_origin      4730..5185
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4817..4836)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5027..5048
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5202..5221)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5321..5343
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5467..5571
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5572..5640,5641..6432)
                     /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"
     rep_origin      6603..7191
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7092..7111
                     /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 gcgtgatcta acatatcctg gtgtggagta ggggacgctg
      181 ctctgacaga ggctcggggg cctgagctgg ctctgtgagc tggggaggag gcagacagcc
      241 aggccttgtc tgcaagcaga cctggcagca ttgggctggc cgccccccag ggcctcctct
      301 tcatgcccag tgaatgactc accttggcac agacacaatg ttcggggtgg gcacagtgcc
      361 tgcttcccgc cgcaccccag cccccctcaa atgccttccg agaagcccat tgagcagggg
      421 gcttgcattg caccccagcc tgacagcctg gcatcttggg ataaaagcag cacagccccc
      481 taggggctgc ccttgctgtg tggcgccacc ggcggtggag aacaaggctc tattcagcct
      541 gtgcccagga aaggggatca ggggatgccc aggcatggac agtgggtggc agggggggag
      601 aggagggctg tctgcttccc agaagtccaa ggacacaaat gggtgagggg agagctctcc
      661 ccatagctgg gctgcggccc aaccccaccc cctcaggcta tgccaggggg tgttgccagg
      721 ggcacccggg catcgccagt ctagcccact ccttcataaa gccctcgcat cccaggagcg
      781 agcagagcca gagcaggttg gagaggagac gcatcacctc cgctgctcgc ggggtctaga
      841 gtcgacgcca ccatgacctt gcacaataac agtacaacct cgcctttgtt tccaaacatc
      901 agctcctcct ggatacacag cccctccgat gcagggctgc ccccgggaac cgtcactcat
      961 ttcggcagct acaatgtttc tcgagcagct ggcaatttct cctctccaga cggtaccacc
     1021 gatgaccctc tgggaggtca taccgtctgg caagtggtct tcatcgcttt cttaacgggc
     1081 atcctggcct tggtgaccat catcggcaac atcctggtaa ttgtgtcatt taaggtcaac
     1141 aagcagctga agacggtcaa caactacttc ctcttaagcc tggcctgtgc cgatctgatt
     1201 atcggggtca tttcaatgaa tctgtttacg acctacatca tcatgaatcg atgggcctta
     1261 gggaacttgg cctgtgacct ctggcttgcc attgactgcg tagccagcaa tgcctctgtt
     1321 atgaatcttc tggtcatcag ctttgacaga tacttttcca tcacgaggcc gctcacgtac
     1381 cgagccaaac gaacaacaaa gagagccggt gtgatgatcg gtctggcttg ggtcatctcc
     1441 tttgtccttt gggctcctgc catcttgttc tggcaatact ttgttggaaa gagaactgtg
     1501 cctccgggag agtgcttcat tcagttcctc agtgagccca ccattacttt tggcacagcc
     1561 atcgctggtt tttatatgcc tgtcaccatt atgactattt tatactggag gatctataag
     1621 gaaactgaaa agcgtaccaa agagcttgct ggcctgcaag cctctgggac agaggcagag
     1681 acagaaaact ttgtccaccc cacgggcagt tctcgaagct gcagcagtta cgaacttcaa
     1741 cagcaaagca tgaaacgctc caacaggagg aagtatggcc gctgccactt ctggttcaca
     1801 accaagagct ggaaacccag ctccgagcag atggaccaag accacagcag cagtgacagt
     1861 tggaacaaca atgatgctgc tgcctccctg gagaactccg cctcctccga cgaggaggac
     1921 attggctccg agacgagagc catctactcc atcgtgctca agcttccggg tcacagcacc
     1981 atcctcaact ccaccaagtt accctcatcg gacaacctgc aggtgcctga ggaggagctg
     2041 gggatggtgg acttggagag gaaagccgac aagctgcagg cccagaagag cgtggacgat
     2101 ggaggcagtt ttccaaaaag cttctccaag cttcccatcc agctagagtc agccgtggac
     2161 acagctaaga cttctgacgt caactcctca gtgggtaaga gcacggccac tctacctctg
     2221 tccttcaagg aagccactct ggccaagagg tttgctctga agaccagaag tcagatcact
     2281 aagcggaaaa ggatgtccct ggtcaaggag aagaaagcgg cccagaccct cagtgcgatc
     2341 ttgcttgcct tcatcatcac ttggacccca tacaacatca tggttctggt gaacaccttt
     2401 tgtgacagct gcatacccaa aaccttttgg aatctgggct actggctgtg ctacatcaac
     2461 agcaccgtga accccgtgtg ctatgctctg tgcaacaaaa cattcagaac cactttcaag
     2521 atgctgctgc tgtgccagtg tgacaaaaaa aagaggcgca agcagcagta ccagcagaga
     2581 cagtcggtca tttttcacaa gcgcgcaccc gagcaggcct tgaaggatcc cccggtcgcc
     2641 accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     2701 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     2761 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     2821 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     2881 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     2941 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     3001 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     3061 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     3121 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     3181 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     3241 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     3301 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaagaattc
     3361 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3421 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3481 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3541 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3601 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3661 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3721 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     3781 tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc
     3841 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3901 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3961 tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct
     4021 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
     4081 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
     4141 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
     4201 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
     4261 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
     4321 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
     4381 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
     4441 tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg
     4501 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     4561 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     4621 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     4681 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     4741 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4801 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4861 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4921 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     4981 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5041 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     5101 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     5161 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     5221 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     5281 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     5341 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     5401 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     5461 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5521 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5581 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5641 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5701 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5761 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5821 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5881 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5941 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6001 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6061 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6121 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6181 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6241 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6301 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6361 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6421 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6481 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6541 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6601 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6661 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6721 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6781 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6841 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6901 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6961 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7021 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7081 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7141 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7201 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
