LOCUS       sequence-163144-        5754 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre recombinase-dependent 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 5754)
  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 5754)
  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-Flex-dTomato-Fishell_7
FEATURES             Location/Qualifiers
     source          1..5754
                     /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"
     protein_bind    complement(1042..1075)
                     /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(1126..1159)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1183..1887)
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MAVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQT
                     AKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDG
                     GLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEI
                     HQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGR
                     HHLFLYGMDELYK"
     primer_bind     complement(1266..1289)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1464..1483)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1733..1751
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1884..1893
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1913..1946
                     /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    1997..2030
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2100..2688
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2153..2173)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2571..2582)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2857..2938
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2894..2913)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2948..2967
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3017..3157
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3017..3146
                     /label=AAV2 ITR
     rep_origin      3232..3687
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3319..3338)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3529..3550
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3704..3723)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3823..3845
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3883..3901)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3969..4073
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4074..4142,4143..4934)
                     /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(4292..4311)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5105..5693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5594..5613
                     /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 ttaagaaagg tcgactccgg aataacttcg tataggatac tttatacgaa gttatgcaga
     1081 atggtagctg gattgtagct gctattagca atatgaaacc tcttaataac ttcgtatagc
     1141 atacattata cgaagttatg gcgcgccact agtttacttt tacttgtaca gctcgtccat
     1201 gccgtacagg aacaggtggt ggcggccctc ggagcgctcg tactgttcca cgatggtgta
     1261 gtcctcgttg tgggaggtga tgtccagctt ggtgtccacg tagtagtagc cgggcagttg
     1321 cacgggcttc ttggccatgt agatggtctt gaactccacc aggtagtggc cgccgtcctt
     1381 cagcttcagg gcctggtgga tctcgccctt cagcacgccg tcgcgggggt acaggcgctc
     1441 ggtggaggcc tcccagccca tggtcttctt ctgcattacg gggccgtcgg gggggaagtt
     1501 ggtgccgcgc atcttcacct tgtagatcag cgtgccgtcc tgcagggagg agtcctgggt
     1561 cacggtcacc agaccgccgt cctcgaagtt catcacgcgc tcccacttga agccctcggg
     1621 gaaggacagc ttcttgtaat cggggatgtc ggcggggtgc ttcacgtacg ccttggagcc
     1681 gtacatgaac tggggggaca ggatgtccca ggcgaagggc agggggccgc ccttggtcac
     1741 cttcagcttg gcggtctggg tgccctcgta ggggcggccc tcgccctcgc cctcgatctc
     1801 gaactcgtgg ccgttcatgg agccctccat gcgcaccttg aagcgcatga actctttgat
     1861 gacctcctcg cccttgctca cggccatggt ggctactagt ggtggcgcta gcataacttc
     1921 gtataaagta tcctatacga agttatttgc cttaacccag aaattatcac tgttattctt
     1981 tagaatggtg caaagaataa cttcgtataa tgtatgctat acgaagttat gaattcgata
     2041 tcaagcttat cgatggcgcg ccccacccct gcagggaatt cgatatcaag cttatcgata
     2101 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2161 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2221 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     2281 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     2341 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     2401 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     2461 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     2521 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     2581 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     2641 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag
     2701 cgctgctcga gagatctgtg atagcggcca tcaagctggc cgcgactcta gatcataatc
     2761 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2821 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2881 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2941 tctagttgtg gtttgtccaa actcatcaat gtatcagctt atcgataccg catgcacgtg
     3001 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3061 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3121 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     3181 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     3241 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3301 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3361 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3421 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     3481 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3541 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
     3601 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3661 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     3721 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3781 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3841 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     3901 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3961 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4021 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4081 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4141 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4201 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4261 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4321 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4381 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4441 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4501 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4561 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4621 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4681 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4741 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4801 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4861 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4921 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4981 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5041 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5101 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5161 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5221 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     5281 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5341 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5401 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5461 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5521 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5581 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5641 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5701 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
