LOCUS       sequence-444006-        6557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV vector expressing ChR2-mCherry that is dependent on both Cre 
            and the hDlx enhancer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6557)
  AUTHORS   Wu SJ, Dai M, Yang SP, McCann C, Qiu Y, Marrero GJ, Stogsdill JA, Di
            Bella DJ, Xu Q, Farhi SL, Macosko EZ, Chen F, Fishell G
  TITLE     Pyramidal neurons proportionately alter the identity and survival of
            specific cortical interneuron subtypes
  JOURNAL   bioRxiv 2024.07.20.604399
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: "bioRxiv 
            2024.07.20.604399"
COMMENT     Sequence Label: pAAV_hDlx_DIO_ChR2_mCherry
FEATURES             Location/Qualifiers
     source          1..6557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(253..272)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     522..540
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(695..1621)
                     /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      1618..1627
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1653..1686
                     /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    1737..1770
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1779..2367
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1832..1852)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2250..2261)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2375..2599
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2601..2741
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2601..2730
                     /label=AAV2 ITR
     rep_origin      2816..3271
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2903..2922)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3113..3134
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3288..3307)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3407..3429
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3467..3485)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3553..3657
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3658..3726,3727..4518)
                     /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(3876..3895)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4689..5277
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5178..5197
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5339..5468
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          6211..6343
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     6326..6345
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    complement(6392..6425)
                     /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(6476..6509)
                     /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(6532..6557,1..682))
                     /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"
ORIGIN
        1 cggtggagtg gcggccctcg gcgcgttcgt actgttccac gatggtgtag tcctcgttgt
       61 gggaggtgat gtccaacttg atgttgacgt tgtaggcgcc gggcagctgc acgggcttct
      121 tggccttgta ggtggtcttg acctcagcgt cgtagtggcc gccgtccttc agcttcagcc
      181 tctgcttgat ctcgcccttc agggcgccgt cctcggggta catccgctcg gaggaggcct
      241 cccagcccat ggtcttcttc tgcattacgg ggccgtcgga ggggaagttg gtgccgcgca
      301 gcttcacctt gtagatgaac tcgccgtcct gcagggagga gtcctgggtc acggtcacca
      361 cgccgccgtc ctcgaagttc atcacgcgct cccacttgaa gccctcgggg aaggacagct
      421 tcaagtagtc ggggatgtcg gcggggtgct tcacgtaggc cttggagccg tacatgaact
      481 gaggggacag gatgtcccag gcgaagggca gggggccacc cttggtcacc ttcagcttgg
      541 cggtctgggt gccctcgtag gggcggccct cgccctcgcc ctcgatctcg aactcgtggc
      601 cgttcacgga gccctccatg tgcaccttga agcgcatgaa ctccttgatg atggccatgt
      661 tatcctcctc gcccttgctc acggcggccg ctggcacggc tccggcctcg gcttcgtctt
      721 cgacgagagt ctcgacctcg atctccgttc cgccgatgtt cagtttggtg gttttgcgga
      781 tatctccgtg aatcaatatg tgctcgtgga tcaggacgcg caggtagtgt cccaacaacc
      841 cccaacaatt tttactcatc agatcaataa tcgtgtgacc tacggtggag ccatagacgc
      901 tcaggacgcc aaaaccttcg ggccccaaaa tgaagagaat tgggaacata ccccagctca
      961 cgaaaaacag ccatgccatg ccggtcacga cctggcggca ccgacccttt ggcacagtat
     1021 gataaccctc gatatatgct ttggcggcgt gaaaaaatgt gttcgcgcca tagcacaatc
     1081 caagacaaaa gaagatgact ttaacatagc cggttgccat ggcgctggta gccccccaca
     1141 cgatagtccc gatgtctgag acaaggagtc ccatggttct cctgctgtag tcgttgctca
     1201 ggccggtgag gttgctcagg cggataagga tgacaggaca agtgagcagc cactctgcat
     1261 agcgcagcca ctgcacccgg tgtcctgtgg caaggtagag catagaggga ttcttaaact
     1321 caaaaaagaa ctcgagaatc accttaacca tttcaatggc gcacacatag atctcctccc
     1381 agccgcatgt agatttccag gtttggtagg catagaacat cagcagcaaa atgctgaatc
     1441 ctgctgcaag ccactgcagg acatttgacg cggtctgagc gccgttcgtg ccgcgagatt
     1501 caatccatcc ggcacagtaa cattgatcct cagggaccag gacggacccg ttcaccacca
     1561 caggattagt aacgaacaaa agttcgcgtc cgacggcaga caaagcgccg ccatagtcca
     1621 tggtggcaag ctttactagt ggtggcgcta gcataacttc gtataaagta tcctatacga
     1681 agttatttgc cttaacccag aaattatcac tgttattctt tagaatggtg caaagaataa
     1741 cttcgtataa tgtatgctat acgaagttat ggcgcgccaa tcaacctctg gattacaaaa
     1801 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     1861 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     1921 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     1981 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     2041 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     2101 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     2161 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc
     2221 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     2281 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     2341 ggatctccct ttgggccgcc tccccgcctg gatcctgtgc cttctagttg ccagccatct
     2401 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2461 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2521 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     2581 gatgcggtgg gctctatggc aggaacccct agtgatggag ttggccactc cctctctgcg
     2641 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     2701 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     2761 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     2821 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2881 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2941 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3001 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     3061 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3121 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     3181 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3241 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     3301 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3361 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3421 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     3481 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     3541 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3601 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3661 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3721 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3781 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3841 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3901 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3961 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4021 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4081 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4141 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4201 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4261 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4321 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4381 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4441 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4501 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4561 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4621 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4681 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4741 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4801 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4861 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4921 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4981 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5041 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5101 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5161 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5221 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5281 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc
     5341 tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg
     5401 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     5461 gggttcctgc tagactgcag agggcccgcg gccgcacgcg tttattcaga atgttatgca
     5521 ctcacagtgg tttggcatgc atctggtgaa ttttttttaa cgaaaaatta gtgttggttt
     5581 cgatgtatgg tagcattctc cctaacgtaa tttgaataat tcagcaaagc cccactacca
     5641 gctgtacttc tgcagcctct tccattcttt tcagcattat aattttggtt aattttcaat
     5701 tttaggtcct acgtctctgc aatttgtgta tgaataacag aataatttcc ctcttttgtt
     5761 tcgcctttcc tgttcctgaa tctaaataaa gatggctttt tagtattaaa agtggaagaa
     5821 aattacaggt aattatcttt gacggtaaaa acgctgtaat cagcgggcta catgaaaaat
     5881 tactctaatt atggctgcat ttaagagaat ggaaaaaaac cttcttgtgg ataaaaacct
     5941 taaattgtcc ccaatgtctg cttcaaattg gatggcactg cagctggagg ctttgttcag
     6001 aattgatcct ggggagctac gaacccaaag tttcacagta ggaagctaga tatcgtttaa
     6061 acttcctgca gcccgggctg ggcataaaag tcagggcaga gccatctatt gcttacattt
     6121 gcttctagcc tgcaagtcga ggagcgcagc cttccagaag cagagcgcgg cgccttaagc
     6181 tgcagaagtt ggtcgtgagg cactgggcac gtaagtatca aggttacaag acaggtttaa
     6241 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
     6301 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc
     6361 aattacagct cttaagaaag ctagctccgg aataacttcg tataggatac tttatacgaa
     6421 gttatgcaga atggtagctg gattgtagct gctattagca atatgaaacc tcttaataac
     6481 ttcgtatagc atacattata cgaagttata acgcgccact agtgaattct tttacttgta
     6541 cagctcgtcc atgccgc
//
