LOCUS       sequence-429015-        7875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses hM4Di-mCherry under the control of distalless homeobox 5 
            and 6 (mDlx) enhancer in Cre-recombinase-dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7875)
  AUTHORS   Rangel Guerrero DK, Balueva K, Barayeu U, Baracskay P, Gridchyn I, 
            Nardin M, Roth CN, Wulff P, Csicsvari J
  TITLE     Hippocampal cholecystokinin-expressing interneurons regulate 
            temporal coding and contextual learning.
  JOURNAL   Neuron. 2024 Apr 10:S0896-6273(24)00197-1. doi: 
            10.1016/j.neuron.2024.03.019.
  PUBMED    38636524
REFERENCE   2  (bases 1 to 7875)
  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: "Neuron. 
            2024 Apr 10:S0896-6273(24)00197-1. doi: 
            10.1016/j.neuron.2024.03.019."
COMMENT     Sequence Label: pAAV-Dlx-FLEX-hM4Di-mCherry
FEATURES             Location/Qualifiers
     source          1..7875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(21..54)
                     /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(125..158)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    230..818
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(283..303)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(701..712)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(845..862)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    851..1075
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(1285..1614)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1299..1434
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1599..1619
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1860..2448)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1940..1959)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2619..3410,3411..3479))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDGGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3242..3261
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3480..3584)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3652..3670
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3708..3730)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3830..3849
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4063..4082
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        4372..4390
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4372..4389
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     repeat_region   4411..4540
                     /label=AAV2 ITR
     intron          5302..5434
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5417..5436
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    5499..5532
                     /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    5595..5628
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(5640..6350)
                     /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(5918..5937)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6187..6205
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      6347..6356
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      7817..7826
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gtgtccggac tagccctagt ataacttcgt ataggatact ttatacgaag ttatcattgg
       61 gattcttcct attttgatcc aagcatcacc atcgaccctc tagtccagat ctcaccatcg
      121 acccataact tcgtatagca tacattatac gaagttatgt ccctcgaaga ggttcgaaac
      181 actgaaaagg gcgaattctg cagatgatcc actcgagaag cttatcgata atcaacctct
      241 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
      301 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
      361 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
      421 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
      481 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
      541 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
      601 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
      661 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
      721 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
      781 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc gactcgctga
      841 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
      901 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
      961 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     1021 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     1081 gaggcggaaa gaaccagctg gggctcgact agagcatggc tacgtagata agtagcatgg
     1141 cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg
     1201 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gcggcctcag
     1261 tgagcgagcg agcgcgcaga gctttttgca aaagcctagg cctccaaaaa agcctcctca
     1321 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
     1381 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
     1441 agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     1501 tgctggggag cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg
     1561 catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca
     1621 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     1681 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     1741 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     1801 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1861 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1921 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1981 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2041 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2101 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2161 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2221 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2281 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2341 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2401 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2461 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2521 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2581 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2641 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     2701 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     2761 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     2821 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     2881 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     2941 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3001 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3061 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3121 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3181 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccgcc gtcaatacgg
     3241 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3301 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3361 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3421 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3481 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3541 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3601 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     3661 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     3721 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     3781 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     3841 cagattgtac tgagagtgca ccattcgacg ctctccctta tgcgactcct gcattaggaa
     3901 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa
     3961 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     4021 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     4081 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     4141 gatctggcta gcgatgaccc tgctgattgg ttcgctgacc atttccgggt gcgggacggc
     4201 gttaccagaa actcagaagg ttcgtccaac caaaccgact ctgacggcag tttacgagag
     4261 agatgatagg gtctgcttca gtaagccaga tgctacacaa ttaggcttgt acatattgtc
     4321 gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg
     4381 acactataga atacacggaa ttaattctag ctgcgcgctc gctcgctcac tgaggccgcc
     4441 cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
     4501 cgcagagagg gagtggccaa ctccatcact aggggttcct tgtagttaat gattaacccg
     4561 ccatgctact tatctacgta gccatgctct agcgcgttta attaagaatt ctatacactc
     4621 acagtggttt ggcatatatt tggtgaaatt ttttaaggaa aaattagtgt tggtttcgat
     4681 atatggtagc tttttctcta acataatttg aataattcag caaagcccta ctaccagctg
     4741 tacttctgca gcctcttcca ttctttccag cattataatt ttggttaatt ttcaatttta
     4801 ggtcctacgt ctctgcaatt tgtgtatgaa taacagaata atttccctct tttgtttcgc
     4861 ctttcctgtt cctgaatcta aataaagatg gctttttagt attaaaagtg gaagaaaatt
     4921 acaggtaatt atctttgacg gtaaaaacgc tgtaatcagc gggctacatg aaaaattact
     4981 ctaattatgg ctgcatttaa gagaatggaa aaaaaccttc ttgtggataa aaaccttaaa
     5041 ttgtccccaa tgtctgcttc aaattggatg gcactgcagc tggaggcttt gttcagaatt
     5101 gatcctgggg agctacgaac ccaaagtttc acagtaggaa gcttgatatc gaattcctgc
     5161 agcccgggct gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctagc
     5221 ctgcaggtcg aggagcgcag ccttccagaa gcagagcgcg gcgccttaag ctgcagaagt
     5281 tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta aggagaccaa
     5341 tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt
     5401 cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccagtt caattacagc
     5461 tcttaagaaa ctagaaagta taggaacttc ggatccccat aacttcgtat aaagtatcct
     5521 atacgaagtt atatcaaaat aggaagacca atgcttcacc atcgacccga attgccaagc
     5581 atcaccatcg acccataact tcgtataatg tatgctatac gaagttatga attcggccgc
     5641 tacttgtaca gctcgtccat gccgccggtg gagtggcggc cctcggcgcg ttcgtactgt
     5701 tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca acttgatgtt gacgttgtag
     5761 gcgccgggca gctgcacggg cttcttggcc ttgtaggtgg tcttgacctc agcgtcgtag
     5821 tggccgccgt ccttcagctt cagcctctgc ttgatctcgc ccttcagggc gccgtcctcg
     5881 gggtacatcc gctcggagga ggcctcccag cccatggtct tcttctgcat tacggggccg
     5941 tcggagggga agttggtgcc gcgcagcttc accttgtaga tgaactcgcc gtcctgcagg
     6001 gaggagtcct gggtcacggt caccacgccg ccgtcctcga agttcatcac gcgctcccac
     6061 ttgaagccct cggggaagga cagcttcaag tagtcgggga tgtcggcggg gtgcttcacg
     6121 taggccttgg agccgtacat gaactgaggg gacaggatgt cccaggcgaa gggcaggggg
     6181 ccacccttgg tcaccttcag cttggcggtc tgggtgccct cgtaggggcg gccctcgccc
     6241 tcgccctcga tctcgaactc gtggccgttc acggagccct ccatgtgcac cttgaagcgc
     6301 atgaactcct tgatgatggc catgttatcc tcctcgccct tgctcaccat ggtggcggag
     6361 ctggtggcga ccggtggatc ccgcctggca gtgccgatgt tccgatactg gcacagcagc
     6421 aggtgccgga aggtcttttt aaaggtggcg ttgcacagag catagcaggc agggttgatg
     6481 gtgctgttga cgtagcagag ccagtagcca atggaccaca ccgtgtcagg gatgcagctc
     6541 tggcagaagg tgttcaccag gaccatgacg ttgtagggcg tccaggtgag gatgaaggcc
     6601 agcagaatgg caaagatcgt tcgtgtcact ttgcgctccc gggccgccat ctgccgcttc
     6661 ttgcgcacct ggttgcgagc gatgctggcg aacttgcggg ccacgttggc cgcagggcgc
     6721 atgccagccg gcgtggcagg cacaatctca atggctgtca cacactcatt gcctgtctgc
     6781 ttcgtcacaa tctggatctt ggaccatctg gaggctgggt tgagggcccg cggctgcagg
     6841 ggaggggcgg gcatggcggg cgtggtggcc tctgtggtgg acagctctgt ggctgggcgt
     6901 tccttggtgt tctgggtggc actgcctgag ctggactcat tggaagtgtc cttatcagcc
     6961 acggggcgcg gtggcggtgg cagcgctggc gggggggcct cctccagctt gccattgcgc
     7021 agctcctccc gggcggcctc cccgggcggg ggcttcttga cgctctgctt cattagtggg
     7081 ctcttgagga aggccagcgt cttggctttc ttctccttcg ggccctcggg ccggtgcttg
     7141 tggactcggc tgcgactggc cagggagatg tggatgtaca gcaccgtcat gatgaccaca
     7201 ggcaggtaga agccagcaat ggctgtgcca aaggtcactg ctgggttgga caggaactgg
     7261 atgaagcact ggttgtcggg caccgtccgc ttacccacca caaactgcca gaacaagatg
     7321 gcaggcgccc agagcacgaa ggacagtacc caggcagcag caatcatgag gcctgccatc
     7381 ttggtggtgc gccgggcagg gtaggtgaga ggcttggtga cgcagaagta gcggtcaaag
     7441 ctgatgatga gaaggttcat gacggaggcg ttgctcacca cgcagtccag ggccagccac
     7501 aggtcgcaga ccacggcgcc caggggccag tagcccttga tgatgtacac ggtgtagagg
     7561 ttcatggaga aggcgcctat gatgagatca gcacacgcca ggctgaagag gaagtagttg
     7621 ttgactgtct gcagctgcct gttgaccttg atggacagca tcaccaggat gttgcccacg
     7681 acagtcacca ggctcaggga gcctgtcact gtggcaatga agaccatttc caccgtctca
     7741 tagcgattgt gggatgatga cgtgaccagg cgcacggact gattgcccga gctgccattg
     7801 acaggtgtga agttggccat ggtggcgcta gtgcggccgc ctcgagagat ctgatatcgc
     7861 ggccgcaccg gtcac
//
