LOCUS       sequence-267511-        6705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent viral expression of ChR2(H134R)-mCherry for neuronal 
            excitation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6705)
  AUTHORS   Atasoy D, Aponte Y, Su HH, Sternson SM
  TITLE     A FLEX switch targets Channelrhodopsin-2 to multiple cell types for 
            imaging and long-range circuit mapping.
  JOURNAL   J Neurosci. 2008 Jul 9. 28(28):7025-30.
  PUBMED    18614669
REFERENCE   2  (bases 1 to 6705)
  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: "J Neurosci.
            2008 Jul 9. 28(28):7025-30."
COMMENT     Sequence Label: AAV-FLEX-rev-ChR2(H134R)-mCherry
FEATURES             Location/Qualifiers
     source          1..6705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   51..180
                     /label=AAV2 ITR
     promoter        491..768
                     /label=chicken beta-actin promoter
     primer_bind     1047..1069
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1131..1150
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1185..1218
                     /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    1281..1314
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1344..2054)
                     /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(1424..1447)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1622..1641)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1891..1909
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2051..2060
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2070..2996)
                     /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      2993..3002
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3020..3053)
                     /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(3124..3157)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3199..3787
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3252..3272)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3670..3681)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3830..3951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3841..3860)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3895..3914
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4113..4242
                     /label=AAV2 ITR
     rep_origin      complement(4479..5067)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4559..4578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5238..6029,6030..6098))
                     /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     5861..5880
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6099..6203)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6229..6684)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6366..6387)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6578..6597
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctgcgcgctc
       61 gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg
      121 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      181 tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct aggaagagta
      241 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      301 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      361 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      421 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      481 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
      541 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
      601 gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg
      661 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
      721 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg
      781 cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
      841 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
      901 tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg
      961 ctccgggagg gccctttgtg cggggggagc ggctcggggc tgtccgcggg gggacggctg
     1021 ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag
     1081 agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct
     1141 ggttattgtg ctgtctcatc attttggcaa agaattggat ccccataact tcgtataaag
     1201 tatcctatac gaagttatat caaaatagga agaccaatgc ttcaccatcg acccgaattg
     1261 ccaagcatca ccatcgaccc ataacttcgt ataatgtatg ctatacgaag ttatactagc
     1321 tagagatatc gtcgacggcc gctttacttg tacagctcgt ccatgccgcc ggtggagtgg
     1381 cggccctcgg cgcgctcgta ctgttccacg atggtgtagt cctcgttgtg ggaggtgatg
     1441 tccagcttga tgttgacgtt gtaggcgccg ggcagctgca cgggcttctt ggccttgtag
     1501 gtggtcttga cctcggcgtc gtagtggccg ccgtccttca gcttcagcct ctgcttgatc
     1561 tcgcccttca gggcgccgtc ctcggggtac atccgctcgg aggaggcctc ccagcccatg
     1621 gtcttcttct gcattacggg gccgtcggag gggaagttgg tgccgcgcag cttcaccttg
     1681 tagatgaact cgccgtcctg cagggaggag tcctgggtca cggtcaccac gccgccgtcc
     1741 tcgaagttca tcacgcgctc ccacttgaag ccctcgggga aggacagctt caagtagtcg
     1801 gggatgtcgg cggggtgctt cacgtaggcc ttggagccgt acatgaactg aggggacagg
     1861 atgtcccagg cgaagggcag ggggccgccc ttggtcacct tcagcttggc ggtctgggtg
     1921 ccctcgtagg ggcggccctc gccctcgccc tcgatctcga actcgtggcc gttcacggag
     1981 ccctccatgt gcaccttgaa gcgcatgaac tccttgatga tggccatgtt gtcctcctcg
     2041 cccttgctca ccatggtggc gaccggtgcc acggctccgg cctcggcttc gtcttcgacg
     2101 agagtctcga cctcgatctc cgttccgccg atgttcagtt tggtggtttt gcggatatct
     2161 ccgtgaatca atatgtgctc gtggatcagg acgcgcaggt agtgtcccaa caacccccaa
     2221 caatttttac tcatcagatc aataatcgtg tgacctacgg tggagccata gacgctcagg
     2281 acgccaaaac cttcgggccc caaaatgaag agaattggga acatacccca gctcacgaaa
     2341 aacagccatg ccatgccggt cacgacctgg cggcaccgac cctttggcac agtatgataa
     2401 ccctcgatat atgctttggc ggcgtgaaaa aatgtgttcg cgccatagca caatccaaga
     2461 caaaagaaga tgactttaac atagccggtt gccatggcgc tggtagcccc ccacacgata
     2521 gtcccgatgt ctgagacaag gagtcccatg gttctcctgc tgtagtcgtt gctcaggccg
     2581 gtgaggttgc tcaggcggat aaggatgaca ggacaagtga gcagccactc tgcatagcgc
     2641 agccactgca cccggtgtcc tgtggcaagg tagagcatag agggattctt aaactcaaaa
     2701 aagaactcga gaatcacctt aaccatttca atggcgcaca catagatctc ctcccagccg
     2761 catgtagatt tccaggtttg gtaggcatag aacatcagca gcaaaatgct gaatcctgct
     2821 gcaagccact gcaggacatt tgacgcggtc tgagcgccgt tcgtgccgcg agattcaatc
     2881 catccggcac agtaacattg atcctcaggg accaggacgg acccgttcac caccacagga
     2941 ttagtaacga acaaaagttc gcgtccgacg gcagacaaag cgccgccata gtccatggtg
     3001 gccggtaccg aattctagta taacttcgta taggatactt tatacgaagt tatcattggg
     3061 attcttccta ttttgatcca agcatcacca tcgaccctct agtccagatc tcaccatcga
     3121 cccataactt cgtatagcat acattatacg aagttatgtc cctcgaagag gttcgaattc
     3181 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3241 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3301 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3361 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3421 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3481 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3541 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     3601 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     3661 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     3721 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     3781 tccccgcatc gataccgtcg acccgggcgg ccgcttcgag cagacatgat aagatacatt
     3841 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     3901 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     3961 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
     4021 taaaacctct acaaatgtgg taaaatcgat aaggatcttc ctagagcatg gctacgtaga
     4081 taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac
     4141 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     4201 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcatta atgaatcggc
     4261 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4321 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4381 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4441 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4501 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4561 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4621 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4681 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4741 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4801 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4861 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4921 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4981 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5041 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5101 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5161 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5221 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5281 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5341 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5401 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5461 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5521 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5581 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5641 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5701 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5761 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5821 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5881 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5941 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6001 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6061 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6121 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6181 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     6241 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     6301 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     6361 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     6421 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     6481 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     6541 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     6601 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     6661 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgc
//
