LOCUS       sequence-459552-        6350 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  enhancer AAV for expression of ChrimsonR-tdTomato under the DLX2.0 
            enhancer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6350)
  AUTHORS   Matveev P, Li AJ, Ye Z, Bowen AJ, Opitz-Araya X, Ting JT, Steinmetz 
            NA
  TITLE     Simultaneous mesoscopic measurement and manipulation of mouse 
            cortical activity
  JOURNAL   bioRxiv 2024.11.01.621418
REFERENCE   2  (bases 1 to 6350)
  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 
            2024.11.01.621418"
COMMENT     Sequence Label: AiP12068 -
            pAAV-DLX2.0-minBG-ChrimsonR-tdTomat...
FEATURES             Location/Qualifiers
     source          1..6350
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      608..617
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             614..1663
                     /codon_start=1
                     /product="red light-drivable channelrhodopsin from 
                     Chlamydomonas noctigama (Klapoetke et al., 2014)"
                     /label=ChrimsonR
                     /note="K176R mutant with accelerated closing kinetics"
                     /translation="MAELISSATRSLFAAGGINPWPNPYHHEDMGCGGMTPTGECFSTE
                     WWCDPSYGLSDAGYGYCFVEATGGYLVVGVEKKQAWLHSRGTPGEKIGAQVCQWIAFSI
                     AIALLTFYGFSAWKATCGWEEVYVCCVEVLFVTLEIFKEFSSPATVYLSTGNHAYCLRY
                     FEWLLSCPVILIRLSNLSGLKNDYSKRTMGLIVSCVGMIVFGMAAGLATDWLKWLLYIV
                     SCIYGGYMYFQAAKCYVEANHSVPKGHCRMVVKLMAYAYFASWGSYPILWAVGPEGLLK
                     LSPYANSIGHSICDIIAKEFWTFLAHHLRIKIHEHILIHGDIRKTTKMEIGGEEVEVEE
                     FVEEEDEDTV"
     primer_bind     complement(3177..3197)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3384..3495
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3613..3753
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3613..3742
                     /label=AAV2 ITR
     rep_origin      3828..4283
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3915..3934)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4125..4146
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4300..4319)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4419..4441
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4479..4497)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4565..4669
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4670..4738,4739..5530)
                     /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(4888..4907)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5701..6289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6190..6209
                     /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 gcgtggtacc ctaaataaag atggcttttt agtattaaaa
      181 gtggaagaaa attacaggta attatctttg acggtaaaaa cgctgtaatc agcgggctac
      241 atgaaaaatt actctaatta tggctgcatt taagagaatg gctaaataaa gatggctttt
      301 tagtattaaa agtggaagaa aattacaggt aattatcttt gacggtaaaa acgctgtaat
      361 cagcgggcta catgaaaaat tactctaatt atggctgcat ttaagagaat ggctaaataa
      421 agatggcttt ttagtattaa aagtggaaga aaattacagg taattatctt tgacggtaaa
      481 aacgctgtaa tcagcgggct acatgaaaaa ttactctaat tatggctgca tttaagagaa
      541 tggagctcgg gctgggcata aaagtcaggg cagagccatc tattgcttac atttgcttct
      601 gggatccgcc accatggctg agctgatcag cagcgccacc agatctctgt ttgccgccgg
      661 aggcatcaac ccttggccta acccctacca ccacgaggac atgggctgtg gaggaatgac
      721 acctacaggc gagtgcttca gcaccgagtg gtggtgtgac ccttcttacg gactgagcga
      781 cgccggatac ggatattgct tcgtggaggc cacaggcggc tacctggtcg tgggagtgga
      841 gaagaagcag gcttggctgc acagcagagg cacaccagga gaaaagatcg gcgcccaggt
      901 ctgccagtgg attgctttca gcatcgccat cgccctgctg acattctacg gcttcagcgc
      961 ctggaaggcc acttgcggtt gggaggaggt ctacgtctgt tgcgtcgagg tgctgttcgt
     1021 gaccctggag atcttcaagg agttcagcag ccccgccaca gtgtacctgt ctaccggcaa
     1081 ccacgcctat tgcctgcgct acttcgagtg gctgctgtct tgccccgtga tcctgatcag
     1141 actgagcaac ctgagcggcc tgaagaacga ctacagcaag cggaccatgg gcctgatcgt
     1201 gtcttgcgtg ggaatgatcg tgttcggcat ggccgcagga ctggctaccg attggctcaa
     1261 gtggctgctg tatatcgtgt cttgcatcta cggcggctac atgtacttcc aggccgccaa
     1321 gtgctacgtg gaagccaacc acagcgtgcc taaaggccat tgccgcatgg tcgtgaagct
     1381 gatggcctac gcttacttcg cctcttgggg cagctaccca atcctctggg cagtgggacc
     1441 agaaggactg ctgaagctga gcccttacgc caacagcatc ggccacagca tctgcgacat
     1501 catcgccaag gagttttgga ccttcctggc ccaccacctg aggatcaaga tccacgagca
     1561 catcctgatc cacggcgaca tccggaagac caccaagatg gagatcggag gcgaggaggt
     1621 ggaagtggaa gagttcgtgg aggaggagga cgaggacaca gtggcggcac cggtagtagc
     1681 agtgagtaag ggcgaggaag tgatcaaaga gttcatgcgg tttaaggtga gaatggaagg
     1741 aagcatgaac ggccacgagt tcgaaattga gggagaagga gagggacggc cctacgaggg
     1801 cacccagaca gccaagctga aagtgacaaa gggcgggcct ctgccattcg cttgggacat
     1861 cctgagccca cagtttatgt acggctccaa ggcctatgtg aaacatccag ctgacattcc
     1921 cgattataag aaactgagct tccccgaggg gtttaagtgg gaaagagtga tgaacttcga
     1981 ggacggaggc ctggtgactg tgacccagga cagctccctg caggatggga ccctgatcta
     2041 caaggtgaaa atgagaggga caaattttcc ccctgatgga cctgtgatgc agaagaaaac
     2101 tatgggatgg gaggcctcca ccgaaaggct gtatccacgc gacggggtgc tgaaaggaga
     2161 aatccaccag gctctgaagc tgaaagatgg gggacattac ctggtggagt tcaagacaat
     2221 ctacatggcc aagaaacctg tgcagctgcc aggctactat tacgtggaca caaaactgga
     2281 tatcacttca cacaacgagg actacactat tgtggagcag tatgaacgga gcgaggggag
     2341 acaccatctg ttcctgggcc atgggactgg aagtaccggc tcagggtcta gtggaaccgc
     2401 ctcaagcgag gataacaata tggctgtgat caaagagttc atgaggttta aggtgcgcat
     2461 ggagggcagc atgaatgggc acgaatttga gattgaagga gagggcgaag ggaggcctta
     2521 cgagggcaca cagactgcca agctgaaagt gaccaaggga ggaccactgc ctttcgcttg
     2581 ggatatcctg tctcctcagt ttatgtacgg aagtaaggcc tatgtcaagc atcccgctga
     2641 cattcctgat tacaagaaac tgtctttccc agagggcttt aagtgggaga gagtgatgaa
     2701 ttttgaagat ggaggcctgg tgaccgtgac acaggactcc tctctgcagg atggcactct
     2761 gatctacaaa gtcaaaatgc gcggcaccaa ttttccaccc gatgggcccg tgatgcagaa
     2821 gaaaacaatg gggtgggagg ccagcactga acggctgtat cctagagacg gagtgctgaa
     2881 gggcgaaatc caccaggccc tgaagctgaa agacggcggc cactacctgg tggagttcaa
     2941 aaccatctac atggccaaga aaccagtgca gctgcccggc tattactatg tggacaccaa
     3001 gctggatatc acatcccaca atgaagacta caccattgtg gaacagtatg agaggtctga
     3061 aggacgccac catctgtttc tgtacggcat ggatgagctg tataagtaag aattcgatat
     3121 cataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3181 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3241 cgtatggctt tcattttctc ctccttgtat aaatcctggt tagttcttgc cacggcggaa
     3301 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3361 tccgtggctc gagagatctt cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3421 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3481 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3541 gcaggacagc aagggggagg attgggaaga caatagcagg catgagatct cacgtgcgga
     3601 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     3661 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     3721 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     3781 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     3841 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3901 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3961 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4021 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     4081 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4141 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     4201 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4261 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     4321 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     4381 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     4441 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     4501 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4561 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4621 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4681 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4741 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4801 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4861 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4921 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4981 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5041 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5101 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5161 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5221 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5281 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5341 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5401 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5461 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5521 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5581 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5641 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5701 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5761 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5821 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5881 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5941 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6001 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6061 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6121 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6181 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6241 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6301 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
