LOCUS       sequence-166303-        6919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gi-coupled hM4D 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 6919)
  AUTHORS   Bryan Roth
  TITLE     Roth lab DREADDs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6919)
  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            "Unpublished"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-hM4D(Gi)-mCherry
FEATURES             Location/Qualifiers
     source          1..6919
                     /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      847..856
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2305..2314
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2311..3021
                     /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(2456..2474)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2724..2743
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3046..3634
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3099..3119)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3517..3528)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3666..4142
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3865..3884)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4182..4322
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4182..4311
                     /label=AAV2 ITR
     rep_origin      4397..4852
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4484..4503)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4694..4715
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4869..4888)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4988..5010
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5048..5066)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5134..5238
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5239..5307,5308..6099)
                     /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      6270..6858
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6759..6778
                     /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 ccatggccaa cttcacacct gtcaatggca gctcgggcaa tcagtccgtg
      901 cgcctggtca cgtcatcatc ccacaatcgc tatgagacgg tggaaatggt cttcattgcc
      961 acagtgacag gctccctgag cctggtgact gtcgtgggca acatcctggt gatgctgtcc
     1021 atcaaggtca acaggcagct gcagacagtc aacaactact tcctcttcag cctggcgtgt
     1081 gctgatctca tcataggcgc cttctccatg aacctctaca ccgtgtacat catcaagggc
     1141 tactggcccc tgggcgccgt ggtctgcgac ctgtggctgg ccctggactg cgtggtgagc
     1201 aacgcctccg tcatgaacct tctcatcatc agctttgacc gctacttctg cgtcaccaag
     1261 cctctcacct accctgcccg gcgcaccacc aagatggcag gcctcatgat tgctgctgcc
     1321 tgggtactgt ccttcgtgct ctgggcgcct gccatcttgt tctggcagtt tgtggtgggt
     1381 aagcggacgg tgcccgacaa ccagtgcttc atccagttcc tgtccaaccc agcagtgacc
     1441 tttggcacag ccattgctgg cttctacctg cctgtggtca tcatgacggt gctgtacatc
     1501 cacatctccc tggccagtcg cagccgagtc cacaagcacc ggcccgaggg cccgaaggag
     1561 aagaaagcca agacgctggc cttcctcaag agcccactaa tgaagcagag cgtcaagaag
     1621 cccccgcccg gggaggccgc ccgggaggag ctgcgcaatg gcaagctgga ggaggccccc
     1681 ccgccagcgc tgccaccgcc accgcgcccc gtggctgata aggacacttc caatgagtcc
     1741 agctcaggca gtgccaccca gaacaccaag gaacgcccag ccacagagct gtccaccaca
     1801 gaggccacca cgcccgccat gcccgcccct cccctgcagc cgcgggccct caacccagcc
     1861 tccagatggt ccaagatcca gattgtgacg aagcagacag gcaatgagtg tgtgacagcc
     1921 attgagattg tgcctgccac gccggctggc atgcgccctg cggccaacgt ggcccgcaag
     1981 ttcgccagca tcgctcgcaa ccaggtgcgc aagaagcggc agatggcggc ccgggagcgc
     2041 aaagtgacac gaacgatctt tgccattctg ctggccttca tcctcacctg gacgccctac
     2101 aacgtcatgg tcctggtgaa caccttctgc cagagctgca tccctgacac ggtgtggtcc
     2161 attggctact ggctctgcta cgtcaacagc accatcaacc ctgcctgcta tgctctgtgc
     2221 aacgccacct ttaaaaagac cttccggcac ctgctgctgt gccagtatcg gaacatcggc
     2281 actgccaggc gggatccacc ggtcgccacc atggtgagca agggcgagga ggataacatg
     2341 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     2401 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     2461 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     2521 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
     2581 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     2641 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     2701 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     2761 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     2821 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     2881 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
     2941 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     3001 atggacgagc tgtacaagta agaattcgat atcaagctta tcgataatca acctctggat
     3061 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3121 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3181 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     3241 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     3301 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3361 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3421 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc
     3481 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     3541 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     3601 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga
     3661 tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca
     3721 ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg
     3781 tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga
     3841 agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat
     3901 cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg
     3961 agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga
     4021 gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc
     4081 caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc
     4141 ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     4201 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     4261 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     4321 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     4381 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4441 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     4501 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4561 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     4621 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4681 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     4741 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     4801 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     4861 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     4921 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4981 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     5041 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     5101 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5161 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5221 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5281 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5341 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5401 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5461 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5521 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5581 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5641 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5701 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5761 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5821 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5881 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5941 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6001 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6061 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6121 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6181 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6241 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6301 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6361 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6421 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6481 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6541 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6601 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6661 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6721 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6781 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6841 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6901 tggccttttg ctcacatgt
//
