LOCUS       sequence-160320-        5760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre recombinase-dependent GFP 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 5760)
  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 5760)
  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-GFP-Fishell_6
FEATURES             Location/Qualifiers
     source          1..5760
                     /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(join(1174..1887,1888..1890,1891..1893))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1177..1887,1888..1890,1891..1893))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1219..1240)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1568..1587
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1827..1848
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1890..1899
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1919..1952
                     /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    2003..2036
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2106..2694
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2159..2179)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2577..2588)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2863..2944
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2900..2919)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2954..2973
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3023..3163
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3023..3152
                     /label=AAV2 ITR
     rep_origin      3238..3693
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3325..3344)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3535..3556
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3710..3729)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3829..3851
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3889..3907)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3975..4079
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4080..4148,4149..4940)
                     /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(4298..4317)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5111..5699
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5600..5619
                     /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 agtttacttg tacagctcgt ccatgccgag
     1201 agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg
     1261 gtctttgctc agggcggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc
     1321 gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat
     1381 gttgtggcgg atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata
     1441 gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt
     1501 gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc
     1561 ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc
     1621 ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa
     1681 gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc
     1741 ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc
     1801 ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac
     1861 cccggtgaac agctcctcgc ccttgctcac catggtggct actagtggtg gcgctagcat
     1921 aacttcgtat aaagtatcct atacgaagtt atttgcctta acccagaaat tatcactgtt
     1981 attctttaga atggtgcaaa gaataacttc gtataatgta tgctatacga agttatgaat
     2041 tcgatatcaa gcttatcgat ggcgcgcccc acccctgcag ggaattcgat atcaagctta
     2101 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2161 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2221 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2281 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2341 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     2401 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     2461 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     2521 tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     2581 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     2641 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     2701 accgagcgct gctcgagaga tctgtgatag cggccatcaa gctggccgcg actctagatc
     2761 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     2821 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     2881 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2941 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cagcttatcg ataccgcatg
     3001 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     3061 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     3121 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     3181 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     3241 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3301 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3361 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3421 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     3481 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3541 tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag
     3601 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3661 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     3721 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     3781 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     3841 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     3901 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     3961 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4021 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4081 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4141 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4201 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4261 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4321 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4381 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4441 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4501 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4561 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4621 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4681 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4741 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4801 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4861 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4921 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4981 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5041 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5101 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5161 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5221 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     5281 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5341 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5401 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5461 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5521 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5581 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5641 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5701 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
