LOCUS       sequence-242471-        6080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GCaMP6f expression in forebrain GABA-ergic interneurons under the 
            control of the mDlx enhancer element.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6080)
  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 6080)
  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-mDlx-GCaMP6f-Fishell-2
FEATURES             Location/Qualifiers
     source          1..6080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          849..981
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     964..983
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1027..1036
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1033..1041,1042..1059,1060..1062,1063..1095,1096..1098,
                     1099..1122,1123..1149,1150..1206,1207..1212,1213..1482,
                     1483..1500,1501..1935,1936..1941,1942..2385)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      1129..1138
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1419..1440
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1546..1567)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1807..1826)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2426..3014
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2479..2499)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2897..2908)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3183..3264
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3220..3239)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3274..3293
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3343..3483
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3343..3472
                     /label=AAV2 ITR
     rep_origin      3558..4013
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3645..3664)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3855..3876
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4030..4049)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4149..4171
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4209..4227)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4295..4399
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4400..4468,4469..5260)
                     /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(4618..4637)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5431..6019
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5920..5939
                     /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 gcgtttaatt aagaattcta tacactcaca gtggtttggc
      181 atatatttgg tgaaattttt taaggaaaaa ttagtgttgg tttcgatata tggtagcttt
      241 ttctctaaca taatttgaat aattcagcaa agccctacta ccagctgtac ttctgcagcc
      301 tcttccattc tttccagcat tataattttg gttaattttc aattttaggt cctacgtctc
      361 tgcaatttgt gtatgaataa cagaataatt tccctctttt gtttcgcctt tcctgttcct
      421 gaatctaaat aaagatggct ttttagtatt aaaagtggaa gaaaattaca ggtaattatc
      481 tttgacggta aaaacgctgt aatcagcggg ctacatgaaa aattactcta attatggctg
      541 catttaagag aatggaaaaa aaccttcttg tggataaaaa ccttaaattg tccccaatgt
      601 ctgcttcaaa ttggatggca ctgcagctgg aggctttgtt cagaattgat cctggggagc
      661 tacgaaccca aagtttcaca gtaggaagct tgatatcgaa ttcctgcagc ccgggctggg
      721 cataaaagtc agggcagagc catctattgc ttacatttgc ttctagcctg caggtcgagg
      781 agcgcagcct tccagaagca gagcgcggcg ccttaagctg cagaagttgg tcgtgaggca
      841 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      901 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      961 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taagactagt
     1021 accggtgcca ccatgggttc tcatcatcat catcatcatg gtatggctag catgactggt
     1081 ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatctcgc caccatggtc
     1141 gactcatcac gtcgtaagtg gaataagaca ggtcacgcag tcagagctat aggtcggctg
     1201 agctcactcg agaacgtcta tatcaaggcc gacaagcaga agaacggcat caaggcgaac
     1261 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgcctacca ctaccagcag
     1321 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcgtgcag
     1381 tccaaacttt cgaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1441 accgccgccg ggatcactct cggcatggac gagctgtaca agggcggtac cggagggagc
     1501 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1561 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gtgagggcga tgccacctac
     1621 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1681 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     1741 cagcacgact tcttcaagtc cgccatgccc gaaggctaca tccaggagcg caccatcttc
     1801 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1861 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1921 aagctggagt acaacctgcc ggaccaactg actgaagagc agatcgcaga atttaaagag
     1981 gaattctccc tatttgacaa ggacggggat gggacaataa caaccaagga gctggggacg
     2041 gtgatgcggt ctctggggca gaaccccaca gaagcagagc tgcaggacat gatcaatgaa
     2101 gtagatgccg acggtgacgg cacaatcgac ttccctgagt tcctgacaat gatggcaaga
     2161 aaaatgaaat acagggacac ggaagaagaa attagagaag cgttcggtgt gtttgataag
     2221 gatggcaatg gctacatcag tgcagcagag cttcgccacg tgatgacaaa ccttggagag
     2281 aagttaacag atgaagaggt tgatgaaatg atcagggaag cagacatcga tggggatggt
     2341 caggtaaact acgaagagtt tgtacaaatg atgacagcga agtgaggcgc gcccctgcag
     2401 ggaattcgat atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     2461 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2521 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2581 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     2641 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     2701 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     2761 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     2821 atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc
     2881 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     2941 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     3001 ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctgtgatag cggccatcaa
     3061 gctggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     3121 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3181 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3241 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3301 cagcttatcg ataccgcatg cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     3361 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     3421 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     3481 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     3541 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3601 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     3661 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3721 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     3781 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3841 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     3901 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     3961 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     4021 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     4081 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     4141 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     4201 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     4261 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4321 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4381 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4441 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4501 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4561 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4621 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4681 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4741 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4801 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4861 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4921 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4981 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5041 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5101 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5161 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5221 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5281 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5341 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5401 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5461 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5521 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5581 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5641 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5701 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5761 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5821 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5881 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5941 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6001 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6061 ctggcctttt gctcacatgt
//
