LOCUS       sequence-337180-        6557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for Cre-dependent transgene expression of ChR2-mCherry in
            cortical interneurons under the control of the Dlx enhancer..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6557)
  AUTHORS   Liu X, Dimidschstein J, Fishell G, Carter AG
  TITLE     Hippocampal inputs engage CCK+ interneurons to mediate 
            endocannabinoid-modulated feed-forward inhibition in the prefrontal 
            cortex.
  JOURNAL   Elife. 2020 Oct 9;9. pii: 55267. doi: 10.7554/eLife.55267.
  PUBMED    33034285
REFERENCE   2  (bases 1 to 6557)
  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: "Elife."; 
            date: "2020-10-9"; volume: "9. pii: 55267. doi"; pages: " 
            10.7554/eLife.55267"
COMMENT     Sequence Label: pAAV-VTKD2-ChR2-mCherry
FEATURES             Location/Qualifiers
     source          1..6557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          851..983
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     966..985
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    complement(1032..1065)
                     /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(1116..1149)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1172..1879)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1450..1469)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1719..1737
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(1892..2818)
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      2815..2824
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2850..2883
                     /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    2934..2967
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2976..3564
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3029..3049)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3447..3458)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3572..3796
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3798..3938
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3798..3927
                     /label=AAV2 ITR
     rep_origin      4013..4468
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4100..4119)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4310..4331
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4485..4504)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4604..4626
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4664..4682)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4750..4854
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4855..4923,4924..5715)
                     /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(5073..5092)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5886..6474
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6375..6394
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6536..6557,1..108)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg
       61 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc tagactgcag
      121 agggcccgcg gccgcacgcg tttattcaga atgttatgca ctcacagtgg tttggcatgc
      181 atctggtgaa ttttttttaa cgaaaaatta gtgttggttt cgatgtatgg tagcattctc
      241 cctaacgtaa tttgaataat tcagcaaagc cccactacca gctgtacttc tgcagcctct
      301 tccattcttt tcagcattat aattttggtt aattttcaat tttaggtcct acgtctctgc
      361 aatttgtgta tgaataacag aataatttcc ctcttttgtt tcgcctttcc tgttcctgaa
      421 tctaaataaa gatggctttt tagtattaaa agtggaagaa aattacaggt aattatcttt
      481 gacggtaaaa acgctgtaat cagcgggcta catgaaaaat tactctaatt atggctgcat
      541 ttaagagaat ggaaaaaaac cttcttgtgg ataaaaacct taaattgtcc ccaatgtctg
      601 cttcaaattg gatggcactg cagctggagg ctttgttcag aattgatcct ggggagctac
      661 gaacccaaag tttcacagta ggaagctaga tatcgtttaa acttcctgca gcccgggctg
      721 ggcataaaag tcagggcaga gccatctatt gcttacattt gcttctagcc tgcaagtcga
      781 ggagcgcagc cttccagaag cagagcgcgg cgccttaagc tgcagaagtt ggtcgtgagg
      841 cactgggcac gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg
      901 cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat
      961 ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct cttaagaaag
     1021 ctagctccgg aataacttcg tataggatac tttatacgaa gttatgcaga atggtagctg
     1081 gattgtagct gctattagca atatgaaacc tcttaataac ttcgtatagc atacattata
     1141 cgaagttata acgcgccact agtgaattct tttacttgta cagctcgtcc atgccgccgg
     1201 tggagtggcg gccctcggcg cgttcgtact gttccacgat ggtgtagtcc tcgttgtggg
     1261 aggtgatgtc caacttgatg ttgacgttgt aggcgccggg cagctgcacg ggcttcttgg
     1321 ccttgtaggt ggtcttgacc tcagcgtcgt agtggccgcc gtccttcagc ttcagcctct
     1381 gcttgatctc gcccttcagg gcgccgtcct cggggtacat ccgctcggag gaggcctccc
     1441 agcccatggt cttcttctgc attacggggc cgtcggaggg gaagttggtg ccgcgcagct
     1501 tcaccttgta gatgaactcg ccgtcctgca gggaggagtc ctgggtcacg gtcaccacgc
     1561 cgccgtcctc gaagttcatc acgcgctccc acttgaagcc ctcggggaag gacagcttca
     1621 agtagtcggg gatgtcggcg gggtgcttca cgtaggcctt ggagccgtac atgaactgag
     1681 gggacaggat gtcccaggcg aagggcaggg ggccaccctt ggtcaccttc agcttggcgg
     1741 tctgggtgcc ctcgtagggg cggccctcgc cctcgccctc gatctcgaac tcgtggccgt
     1801 tcacggagcc ctccatgtgc accttgaagc gcatgaactc cttgatgatg gccatgttat
     1861 cctcctcgcc cttgctcacg gcggccgctg gcacggctcc ggcctcggct tcgtcttcga
     1921 cgagagtctc gacctcgatc tccgttccgc cgatgttcag tttggtggtt ttgcggatat
     1981 ctccgtgaat caatatgtgc tcgtggatca ggacgcgcag gtagtgtccc aacaaccccc
     2041 aacaattttt actcatcaga tcaataatcg tgtgacctac ggtggagcca tagacgctca
     2101 ggacgccaaa accttcgggc cccaaaatga agagaattgg gaacataccc cagctcacga
     2161 aaaacagcca tgccatgccg gtcacgacct ggcggcaccg accctttggc acagtatgat
     2221 aaccctcgat atatgctttg gcggcgtgaa aaaatgtgtt cgcgccatag cacaatccaa
     2281 gacaaaagaa gatgacttta acatagccgg ttgccatggc gctggtagcc ccccacacga
     2341 tagtcccgat gtctgagaca aggagtccca tggttctcct gctgtagtcg ttgctcaggc
     2401 cggtgaggtt gctcaggcgg ataaggatga caggacaagt gagcagccac tctgcatagc
     2461 gcagccactg cacccggtgt cctgtggcaa ggtagagcat agagggattc ttaaactcaa
     2521 aaaagaactc gagaatcacc ttaaccattt caatggcgca cacatagatc tcctcccagc
     2581 cgcatgtaga tttccaggtt tggtaggcat agaacatcag cagcaaaatg ctgaatcctg
     2641 ctgcaagcca ctgcaggaca tttgacgcgg tctgagcgcc gttcgtgccg cgagattcaa
     2701 tccatccggc acagtaacat tgatcctcag ggaccaggac ggacccgttc accaccacag
     2761 gattagtaac gaacaaaagt tcgcgtccga cggcagacaa agcgccgcca tagtccatgg
     2821 tggcaagctt tactagtggt ggcgctagca taacttcgta taaagtatcc tatacgaagt
     2881 tatttgcctt aacccagaaa ttatcactgt tattctttag aatggtgcaa agaataactt
     2941 cgtataatgt atgctatacg aagttatggc gcgccaatca acctctggat tacaaaattt
     3001 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3061 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3121 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3181 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3241 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3301 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3361 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     3421 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3481 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3541 tctccctttg ggccgcctcc ccgcctggat cctgtgcctt ctagttgcca gccatctgtt
     3601 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3661 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3721 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3781 gcggtgggct ctatggcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     3841 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     3901 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     3961 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     4021 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4081 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4141 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4201 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     4261 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4321 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     4381 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4441 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     4501 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4561 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4621 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     4681 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4741 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4801 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4861 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4921 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4981 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5041 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5101 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5161 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5221 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5281 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5341 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5401 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5461 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5521 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5581 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5641 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5701 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5761 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5821 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5881 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5941 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6001 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6061 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6121 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6181 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6241 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6301 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6361 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6421 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6481 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcctgc
     6541 aggcagctgc gcgctcg
//
