LOCUS       sequence-477833-        7557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent viral expression of oChIEF-tdTomato for optogenetic 
            excitation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7557)
  AUTHORS   Bell KA, Shim H, Chen CK, McQuiston AR
  TITLE     Nicotinic excitatory postsynaptic potentials in hippocampal CA1 
            interneurons are predominantly mediated by nicotinic receptors that 
            contain alpha4 and beta2 subunits.
  JOURNAL   Neuropharmacology. 2011 Aug 25.
  PUBMED    21878344
REFERENCE   2  (bases 1 to 7557)
  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: 
            "Neuropharmacology. 2011 Aug 25."
COMMENT     Sequence Label: AAV-Flex-rev-oChIEF-tdTomato
FEATURES             Location/Qualifiers
     source          1..7557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     primer_bind     997..1019
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1081..1100
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1135..1168
                     /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    1231..1264
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1282..2712)
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     protein_bind    complement(3822..3855)
                     /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(3926..3959)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4001..4589
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4054..4074)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4472..4483)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4632..4753
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4643..4662)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4697..4716
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4915..5044
                     /label=AAV2 ITR
     rep_origin      complement(5281..5869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5361..5380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6040..6831,6832..6900))
                     /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     6663..6682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6901..7005)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7031..7486)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7168..7189)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7380..7399
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagagta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      421 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
      481 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
      541 gggggcgggg gggggggggg ggggcgcccc gggggggggg gggcggggcg gggggggggg
      601 gggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      661 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      721 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
      781 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      841 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
      901 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggc tgtccgcggg
      961 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1021 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1081 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattggat ccccataact
     1141 tcgtataaag tatcctatac gaagttatat caaaatagga agaccaatgc ttcaccatcg
     1201 acccgaattg ccaagcatca ccatcgaccc ataacttcgt ataatgtatg ctatacgaag
     1261 ttatactagg atcccctcga gctacttgta cagctcgtcc atgccgtaca ggaacaggtg
     1321 gtggcggccc tcggagcgct cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt
     1381 gatgtccagc ttggtgtcca cgtagtagta gccgggcagt tgcacgggct tcttggccat
     1441 gtagatggtc ttgaactcca ccaggtagtg gccgccgtcc ttcagcttca gggcctggtg
     1501 gatctcgccc ttcagcacgc cgtcgcgggg gtacaggcgc tcggtggagg cctcccagcc
     1561 catggtcttc ttctgcatta cggggccgtc gggggggaag ttggtgccgc gcatcttcac
     1621 cttgtagatc agcgtgccgt cctgcaggga ggagtcctgg gtcacggtca ccagaccgcc
     1681 gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcttgta
     1741 atcggggatg tcggcggggt gcttcacgta cgccttggag ccgtacatga actgggggga
     1801 caggatgtcc caggcgaagg gcagggggcc gcccttggtc accttcagct tggcggtctg
     1861 ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcat
     1921 ggagccctcc atgcgcacct tgaagcgcat gaactctttg atgacggcca tgttgttgtc
     1981 ctcggaggag gcggtgccgg agctgccgct gccggtgctg ccggtgccat gccccaggaa
     2041 caggtggtgg cggccctcgg agcgctcgta ctgttccacg atggtgtagt cctcgttgtg
     2101 ggaggtgatg tccagcttgg tgtccacgta gtagtagccg ggcagttgca cgggcttctt
     2161 ggccatgtag atggtcttga actccaccag gtagtggccg ccgtccttca gcttcagggc
     2221 ctggtggatc tcgcccttca gcacgccgtc gcgggggtac aggcgctcgg tggaggcctc
     2281 ccagcccatg gtcttcttct gcattacggg gccgtcgggg gggaagttgg tgccgcgcat
     2341 cttcaccttg tagatcagcg tgccgtcctg cagggaggag tcctgggtca cggtcaccag
     2401 accgccgtcc tcgaagttca tcacgcgctc ccacttgaag ccctcgggga aggacagctt
     2461 cttgtaatcg gggatgtcgg cggggtgctt cacgtacgcc ttggagccgt acatgaactg
     2521 gggggacagg atgtcccagg cgaagggcag ggggccgccc ttggtcacct tcagcttggc
     2581 ggtctgggtg ccctcgtagg ggcggccctc gccctcgccc tcgatctcga actcgtggcc
     2641 gttcatggag ccctccatgc gcaccttgaa gcgcatgaac tctttgatga cctcctcgcc
     2701 cttgctcacc atatcgatgc tgctctcgta cttgccggtg cccttgttca cggcgccggc
     2761 ctcggcctcg tcctccacca gggtctccac ctcgatctcg gtgccgccga tgttcagctt
     2821 ggtggtcttt ctgatgtcgc cgtggatcag gatgtgctcg tggatcagca ctctcaggta
     2881 gtggcccagc aggccccagc agttcttgct catcaggtcg atgatggtgt ggcccacggt
     2941 gctgccgtac acgctcagca cgccgaagcc ctcggggccc aggatgaaca ggatggggaa
     3001 catgccccag ctcacgaaga acagccaggc catgccggtc accacctgtc tgcatctgcc
     3061 cttgggcacg gtgtggtagg cctcgatgta caccttggcg gcgttgaaga aggtgtagat
     3121 gccgtagcac aggcccatca ggaagaagat cactctcacg tagcccttgc tcagggcggc
     3181 ggtggtgccc cacacgatgg tgccgatgtc gctcaccagc aggcccatgg ttctcttgtt
     3241 gtagtcgttg gccaggccgg tcaggttgct caggtggatc aggaccacgg ggcaggtcag
     3301 cagccactcg gcgtatctca gccacacggt cttgttgccg ttgctgctgt agatcacggc
     3361 gggctcgtcg aactcgtgga agtactctat gatgaacttg atcatctcga tggtggccac
     3421 gtagatctcc tcccagccgc aggtactctt ccaggtctgg tagccgtaga acatcaggca
     3481 cagggcgctc agggcgaagg tgatccactg caggatgttg gcggccagct tctcggcgtt
     3541 ggtgccgtta ctcttcagcc aggccaggca gaagcactgg ccgttgttgg ggatgcagat
     3601 cacgctgccg ttgttctcca gggtgtagct ggtctggaac agcattctct cgtgggctct
     3661 gtggaacacg tagtcggggc cgtcctgggt ggccacgggc acggtggcgt cgctgccggt
     3721 gctggcgccg gcgctgccgg cggccagggc cacggccagg gccagggcca gcagccaggg
     3781 tcttctgctc accatggtca ccggtggtac cgaattctag tataacttcg tataggatac
     3841 tttatacgaa gttatcattg ggattcttcc tattttgatc caagcatcac catcgaccct
     3901 ctagtccaga tctcaccatc gacccataac ttcgtatagc atacattata cgaagttatg
     3961 tccctcgaag aggttcgaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
     4021 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     4081 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     4141 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     4201 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     4261 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     4321 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     4381 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     4441 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     4501 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     4561 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacccgggc ggccgcttcg
     4621 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     4681 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     4741 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     4801 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct
     4861 tcctagagca tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac
     4921 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     4981 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     5041 gcagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5101 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5161 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5221 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5281 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5341 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5401 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5461 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5521 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5581 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5641 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5701 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5761 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5821 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5881 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5941 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6001 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6061 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6121 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6181 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6241 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6301 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6361 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6421 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6481 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6541 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6601 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6661 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6721 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6781 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6841 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6901 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6961 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7021 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     7081 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     7141 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     7201 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     7261 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     7321 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     7381 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     7441 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     7501 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccag
//
