LOCUS       sequence-162264-        6781 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Double floxed Gs-coupled rM3D DREADD fused with mCherry under the 
            control of human synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6781)
  AUTHORS   Bryan Roth
  TITLE     Roth lab DREADDs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6781)
  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            "Unpublished"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-hSyn-DIO-rM3D(Gs)-mCherry
FEATURES             Location/Qualifiers
     source          1..6781
                     /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        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(660..693)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(744..777)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(786..1496)
                     /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(1064..1083)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1333..1351
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1493..1502
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2766..2799
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2850..2883
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2908..3496
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2961..2981)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3379..3390)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3528..4004
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3727..3746)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4044..4184
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4044..4173
                     /label=AAV2 ITR
     rep_origin      4259..4714
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4346..4365)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4556..4577
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4731..4750)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4850..4872
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4910..4928)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4996..5100
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5101..5169,5170..5961)
                     /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(5319..5338)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6132..6720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6621..6640
                     /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gactccggaa
      661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
      721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
      781 gcgccttact tgtacagctc gtccatgccg ccggtggagt ggcggccctc ggcgcgttcg
      841 tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccaactt gatgttgacg
      901 ttgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt gacctcagcg
      961 tcgtagtggc cgccgtcctt cagcttcagc ctctgcttga tctcgccctt cagggcgccg
     1021 tcctcggggt acatccgctc ggaggaggcc tcccagccca tggtcttctt ctgcattacg
     1081 gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc
     1141 tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt catcacgcgc
     1201 tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc
     1261 ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca ggcgaagggc
     1321 agggggccac ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc
     1381 tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat gtgcaccttg
     1441 aagcgcatga actccttgat gatggccatg ttatcctcct cgcccttgct caccatggtg
     1501 gcgaccggtg gatccgccaa ggcctgctcc ggcactcgct tgtgaaaaat gaccgactgt
     1561 ctctgctggt actgctgttt gcgcctcttc cttttgtcac actggcacaa gaggagcgtc
     1621 ttgaaggtgg ttctgaatgt tttgttgcac agggcatagc acacagggtt cacggtgctg
     1681 ttgatatagc acagccagta gcccagattc caataggttt tgggtatgca gctgtcacag
     1741 aaggtgttca ccaggaccat gatgttgtag ggggtccacg tgatgatgaa ggccagcaag
     1801 atggcactga gcgtctgcag agctttgtgt tccctcatgg ccatgacacg ggacgtcttc
     1861 ctcttgctgg cacggccgtt gccgaggatg ggctggtgtt gggggagcgg gggtggctgc
     1921 ggctgctcct ggctgccata gaaccggccc tcgcagcggt cgatcttcct gatctgctcc
     1981 ttggcctccc ggtacacccg ccagtataaa atagtcatga tggtgacagg catgtaaaag
     2041 ccagcgatcg ccgtgccgaa ggtgatggtg ggctcactca gaaactgaat gaaacattct
     2101 cctgggggca cagttctctt ccctacaaag tattgccaga acaagatggc aggagcccat
     2161 aggacaaagg agatgaccca agccagacca atcatcacac cagctcgagc cctggtcatc
     2221 aggctctggt agcggaatgg agaagtgatg gaaaagtacc tgtcaaagct gatgaccagc
     2281 agattcatga cagaggcatt gctggccaca cagtcaatgg agagccagag gtcgcaggct
     2341 aagttcccca gtgcccaacg gttcataatg atgtaggtag tgaacaggtt catggaaatg
     2401 accccgatga tcaggtctgc acaggccagg cttaagagga agtagttgtt gactgtcttc
     2461 agctgtttgt tgaccttgaa ggccacaatg acaaggatgt tgccaatgat ggtcaccaat
     2521 gccaggaagc cagttaagaa ggcaatgaag accacttgcc agatggtgtg acccccgaga
     2581 gggtcgctgg aggtgtcgtt tgaggagaaa ttcccagttt cttgtgaaat gttgtagctg
     2641 cccaactgag tgactgtccc caagggcagc cctgcctccg agggactgtg cacccaggaa
     2701 gagctgatgt tgggaaacaa aggcgaggtt gtactgttac tgtgcaaggt catggtggcg
     2761 ctagcataac ttcgtataaa gtatcctata cgaagttatt tgccttaacc cagaaattat
     2821 cactgttatt ctttagaatg gtgcaaagaa taacttcgta taatgtatgc tatacgaagt
     2881 tatgaattcg atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
     2941 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     3001 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3061 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     3121 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     3181 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     3241 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     3301 aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg
     3361 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     3421 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     3481 tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct
     3541 gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc
     3601 ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta
     3661 tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct
     3721 gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct
     3781 ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg
     3841 catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat
     3901 tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat
     3961 tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa
     4021 ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     4081 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     4141 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     4201 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     4261 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4321 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4381 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4441 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     4501 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4561 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     4621 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4681 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     4741 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     4801 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     4861 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     4921 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     4981 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5041 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5101 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5161 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5221 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5281 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5341 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5401 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5461 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5521 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5581 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5641 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5701 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5761 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5821 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5881 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5941 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6001 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6061 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6121 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6181 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6241 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6301 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6361 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6421 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6481 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6541 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6601 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6661 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6721 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6781 t
//
