LOCUS       sequence-160319-        6999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gi-DREADD (P2A) nuclear dTomato expression in forebrain GABA-ergic 
            interneurons under the control of the hDlx enhancer element.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6999)
  AUTHORS   Dimidschstein J, Chen Q, Tremblay R, Rogers SL, Saldi GA, Guo L, Xu 
            Q, Liu R, Lu C, Chu J, Avery MC, Rashid MS, Baek M, Jacob AL, Smith 
            GB, Wilson DE, Kosche G, Kruglikov I, Rusielewicz T, Kotak VC, 
            Mowery TM, Anderson SA, Callaway EM, Dasen JS, Fitzpatrick D, 
            Fossati V, Long MA, Noggle S, Reynolds JH, Sanes DH, Rudy B, Feng G,
            Fishell G
  TITLE     A viral strategy for targeting and manipulating interneurons across 
            vertebrate species.
  JOURNAL   Nat Neurosci. 2016 Oct 31. doi: 10.1038/nn.4430.
  PUBMED    27798629
REFERENCE   2  (bases 1 to 6999)
  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: "Nat 
            Neurosci. 2016 Oct 31. doi: 10.1038/nn.4430."
COMMENT     Sequence Label: pAAV-hDlx-GiDREADD-dTomato-Fishell-5
FEATURES             Location/Qualifiers
     source          1..6999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     intron          860..992
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     975..994
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1035..1044
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2502..2558
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2571..2591
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2592..3290
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLYGMDELYK"
     primer_bind     complement(2722..2740)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2990..3009
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3184..3207
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    3345..3933
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3398..3418)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3816..3827)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4102..4183
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4139..4158)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4193..4212
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4262..4402
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4262..4391
                     /label=AAV2 ITR
     rep_origin      4477..4932
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4564..4583)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4774..4795
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4949..4968)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5068..5090
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5128..5146)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5214..5318
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5319..5387,5388..6179)
                     /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(5537..5556)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6350..6938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6839..6858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtttatt cagaatgtta tgcactcaca
      181 gtggtttggc atgcatctgg tgaatttttt ttaacgaaaa attagtgttg gtttcgatgt
      241 atggtagcat tctccctaac gtaatttgaa taattcagca aagccccact accagctgta
      301 cttctgcagc ctcttccatt cttttcagca ttataatttt ggttaatttt caattttagg
      361 tcctacgtct ctgcaatttg tgtatgaata acagaataat ttccctcttt tgtttcgcct
      421 ttcctgttcc tgaatctaaa taaagatggc tttttagtat taaaagtgga agaaaattac
      481 aggtaattat ctttgacggt aaaaacgctg taatcagcgg gctacatgaa aaattactct
      541 aattatggct gcatttaaga gaatggaaaa aaaccttctt gtggataaaa accttaaatt
      601 gtccccaatg tctgcttcaa attggatggc actgcagctg gaggctttgt tcagaattga
      661 tcctggggag ctacgaaccc aaagtttcac agtaggaagc ttgatatcga attcctgcag
      721 cccgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg cttctagcct
      781 gcaggtcgag gagcgcagcc ttccagaagc agagcgcggc gccttaagct gcagaagttg
      841 gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata
      901 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct
      961 tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca attacagctc
     1021 ttaagaaagc tagcgccacc atggccaact tcacacctgt caatggcagc tcgggcaatc
     1081 agtccgtgcg cctggtcacg tcatcatccc acaatcgcta tgagacggtg gaaatggtct
     1141 tcattgccac agtgacaggc tccctgagcc tggtgactgt cgtgggcaac atcctggtga
     1201 tgctgtccat caaggtcaac aggcagctgc agacagtcaa caactacttc ctcttcagcc
     1261 tggcgtgtgc tgatctcatc ataggcgcct tctccatgaa cctctacacc gtgtacatca
     1321 tcaagggcta ctggcccctg ggcgccgtgg tctgcgacct gtggctggcc ctggactgcg
     1381 tggtgagcaa cgcctccgtc atgaaccttc tcatcatcag ctttgaccgc tacttctgcg
     1441 tcaccaagcc tctcacctac cctgcccggc gcaccaccaa gatggcaggc ctcatgattg
     1501 ctgctgcctg ggtactgtcc ttcgtgctct gggcgcctgc catcttgttc tggcagtttg
     1561 tggtgggtaa gcggacggtg cccgacaacc agtgcttcat ccagttcctg tccaacccag
     1621 cagtgacctt tggcacagcc attgctggct tctacctgcc tgtggtcatc atgacggtgc
     1681 tgtacatcca catctccctg gccagtcgca gccgagtcca caagcaccgg cccgagggcc
     1741 cgaaggagaa gaaagccaag acgctggcct tcctcaagag cccactaatg aagcagagcg
     1801 tcaagaagcc cccgcccggg gaggccgccc gggaggagct gcgcaatggc aagctggagg
     1861 aggccccccc gccagcgctg ccaccgccac cgcgccccgt ggctgataag gacacttcca
     1921 atgagtccag ctcaggcagt gccacccaga acaccaagga acgcccagcc acagagctgt
     1981 ccaccacaga ggccaccacg cccgccatgc ccgcccctcc cctgcagccg cgggccctca
     2041 acccagcctc cagatggtcc aagatccaga ttgtgacgaa gcagacaggc aatgagtgtg
     2101 tgacagccat tgagattgtg cctgccacgc cggctggcat gcgccctgcg gccaacgtgg
     2161 cccgcaagtt cgccagcatc gctcgcaacc aggtgcgcaa gaagcggcag atggcggccc
     2221 gggagcgcaa agtgacacga acgatctttg ccattctgct ggccttcatc ctcacctgga
     2281 cgccctacaa cgtcatggtc ctggtgaaca ccttctgcca gagctgcatc cctgacacgg
     2341 tgtggtccat tggctactgg ctctgctacg tcaacagcac catcaaccct gcctgctatg
     2401 ctctgtgcaa cgccaccttt aaaaagacct tccggcacct gctgctgtgc cagtatcgga
     2461 acatcggcac tgccaggcgg tccggagata acggaagcgg agccactaac ttctccctgt
     2521 tgaaacaagc aggggatgtc gaagagaatc ccgggccatc tagaatggcc cccaagaaga
     2581 agaggaaggt ggtgagcaag ggcgaggagg tcatcaaaga gttcatgcgc ttcaaggtgc
     2641 gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     2701 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg
     2761 cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg aagcaccccg
     2821 ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     2881 tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg caggacggca
     2941 cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc cccgtaatgc
     3001 agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc
     3061 tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac ctggtggagt
     3121 tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac tacgtggaca
     3181 ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgct
     3241 ccgagggccg ccaccacctg ttcctgtacg gcatggacga gctgtacaag taagtcgacg
     3301 gcgcgcccca cccctgcagg gaattcgata tcaagcttat cgataatcaa cctctggatt
     3361 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3421 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3481 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3541 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3601 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     3661 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3721 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct
     3781 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3841 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3901 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat
     3961 ctgtgatagc ggccatcaag ctggccgcga ctctagatca taatcagcca taccacattt
     4021 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     4081 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     4141 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4201 tccaaactca tcaatgtatc agcttatcga taccgcatgc acgtgcggac cgagcggccg
     4261 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     4321 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     4381 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     4441 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     4501 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4561 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4621 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4681 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4741 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4801 cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct
     4861 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4921 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4981 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5041 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5101 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5161 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5221 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5281 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5341 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5401 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5461 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5521 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5581 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5641 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5701 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5761 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5821 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5881 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5941 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6001 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6061 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6121 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6181 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6241 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6301 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6361 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6421 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6481 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6541 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6601 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6661 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6721 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6781 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6841 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6901 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6961 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
