LOCUS       sequence-475760-        7798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ChETA-tdTomato in cells lacking Cre (Cre-Off, flanked by 
            alternative lox FAS sites), for optogenetic excitation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7798)
  AUTHORS   Saunders A, Johnson CA, Sabatini BL
  TITLE     Novel recombinant adeno-associated viruses for Cre activated and 
            inactivated transgene expression in neurons.
  JOURNAL   Front Neural Circuits. 2012 Jul 27;6:47. doi: 
            10.3389/fncir.2012.00047. eCollection 2012.
  PUBMED    22866029
REFERENCE   2  (bases 1 to 7798)
  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: "Front 
            Neural Circuits."; date: "2012-07-27"; volume: "6:47. doi"; pages: "
            10.3389/fncir.2012.00047. eCollection 2012"
COMMENT     Sequence Label: pAAV-Ef1a-FAS-ChETA-TdTomato-WPRE-pA
FEATURES             Location/Qualifiers
     source          1..7798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     525..544
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(763..867)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     935..953
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(991..1013)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1113..1132
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1149..1604)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1286..1307)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1498..1517
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1679..1819
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1690..1819
                     /label=AAV2 ITR
     polyA_signal    1859..2335
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     2117..2136
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    2367..2955
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2473..2484
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2882..2902
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3020..4450)
                     /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"
     regulatory      4447..4456
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4466..5392)
                     /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
                     PSMLYLATGHRVQWLRYATWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      5389..5398
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5471..6649)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5480..6418
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5503..5523)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     repeat_region   6750..6879
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(6941..7529)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7021..7040)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7700..7798,1..693,694..762))
                     /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"
ORIGIN
        1 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
       61 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
      121 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
      181 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
      241 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
      301 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
      361 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
      421 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
      481 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
      541 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
      601 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
      661 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
      721 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
      781 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
      841 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
      901 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
      961 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     1021 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     1081 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     1141 gcaccataaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     1201 cagctcattt tttaaccaat agaccgaaat cggcaaaatc ccttataaat caaaagaata
     1261 gcccgagata gagttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     1321 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     1381 atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     1441 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     1501 gaagaaagcg aaaggagcgg gcgctaaggc gctggcaagt gtagcggtca cgctgcgcgt
     1561 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg gttgctttga
     1621 cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgcccc
     1681 tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg
     1741 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac
     1801 tccatcacta ggggttcctg cggccgctcg gtccgcacgt ggttacctac aaaatcagaa
     1861 ggacagggaa gggagcagtg gttcacgcct gtaatcccag caatttggga ggccaaggtg
     1921 ggtagatcac ctgagattag gagttggaga ccagcctggc caatatggtg aaaccccgtc
     1981 tctaccaaaa aaacaaaaat tagctgagcc tggtcatgca tgcctggaat cccaacaact
     2041 cgggaggctg aggcaggaga atcgcttgaa cccaggaggc ggagattgca gtgagccaag
     2101 attgtgccac tgcactccag cttggttccc aatagacccc gcaggcccta caggttgtct
     2161 tcccaacttg ccccttgctc cataccaccc ccctccaccc cataatatta tagaaggaca
     2221 cctagtcaga caaaatgatg caacttaatt ttattaggac aaggctggtg ggcactggag
     2281 tggcaacttc cagggccagg agaggcactg gggaggggtc acagggatgc cacccgtaga
     2341 tctctcgagc agcgctcggt atcgatgcgg ggaggcggcc caaagggaga tccgactcgt
     2401 ctgagggcga aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg
     2461 aaggtccgct ggattgaggg ccgaagggac gtagcagaag gacgtcccgc gcagaatcca
     2521 ggtggcaaca taggcgagca gccaaggaaa ggacgatgat ttccccgaca acaccacgga
     2581 attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag
     2641 ttccgccgtg gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt
     2701 ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg
     2761 cctgacaacg ggccacaact cctcataaag agacagcaac caggatttat acaaggagga
     2821 gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc
     2881 acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta
     2941 atccagaggt tgattatcga taagcttgat atcgaattca caacttcgta tagaaaggta
     3001 tatacgaagt tgtgctagct tacttgtaca gctcgtccat gccgtacagg aacaggtggt
     3061 ggcggccctc ggagcgctcg tactgttcca cgatggtgta gtcctcgttg tgggaggtga
     3121 tgtccagctt ggtgtccacg tagtagtagc cgggcagttg cacgggcttc ttggccatgt
     3181 agatggtctt gaactccacc aggtagtggc cgccgtcctt cagcttcagg gcctggtgga
     3241 tctcgccctt cagcacgccg tcgcgggggt acaggcgctc ggtggaggcc tcccagccca
     3301 tggtcttctt ctgcattacg gggccgtcgg gggggaagtt ggtgccgcgc atcttcacct
     3361 tgtagatcag cgtgccgtcc tgcagggagg agtcctgggt cacggtcacc agaccgccgt
     3421 cctcgaagtt catcacgcgc tcccacttga agccctcggg gaaggacagc ttcttgtaat
     3481 cggggatgtc ggcggggtgc ttcacgtacg ccttggagcc gtacatgaac tggggggaca
     3541 ggatgtccca ggcgaagggc agggggccgc ccttggtcac cttcagcttg gcggtctggg
     3601 tgccctcgta ggggcggccc tcgccctcgc cctcgatctc gaactcgtgg ccgttcatgg
     3661 agccctccat gcgcaccttg aagcgcatga actctttgat gacggccatg ttgttgtcct
     3721 cggaggaggc ggtgccggag ctgccgctgc cggtgctgcc ggtgccatgc cccaggaaca
     3781 ggtggtggcg gccctcggag cgctcgtact gttccacgat ggtgtagtcc tcgttgtggg
     3841 aggtgatgtc cagcttggtg tccacgtagt agtagccggg cagttgcacg ggcttcttgg
     3901 ccatgtagat ggtcttgaac tccaccaggt agtggccgcc gtccttcagc ttcagggcct
     3961 ggtggatctc gcccttcagc acgccgtcgc gggggtacag gcgctcggtg gaggcctccc
     4021 agcccatggt cttcttctgc attacggggc cgtcgggggg gaagttggtg ccgcgcatct
     4081 tcaccttgta gatcagcgtg ccgtcctgca gggaggagtc ctgggtcacg gtcaccagac
     4141 cgccgtcctc gaagttcatc acgcgctccc acttgaagcc ctcggggaag gacagcttct
     4201 tgtaatcggg gatgtcggcg gggtgcttca cgtacgcctt ggagccgtac atgaactggg
     4261 gggacaggat gtcccaggcg aagggcaggg ggccgccctt ggtcaccttc agcttggcgg
     4321 tctgggtgcc ctcgtagggg cggccctcgc cctcgccctc gatctcgaac tcgtggccgt
     4381 tcatggagcc ctccatgcgc accttgaagc gcatgaactc tttgatgacc tcctcgccct
     4441 tgctcaccat ggtggcggcc gctggcacgg ctccggcctc ggcttcgtct tcgacgagag
     4501 tctcgacctc gatctccgtt ccgccgatgt tcagtttggt ggttttgcgg atatctccgt
     4561 gaatcaatat gtgctcgtgg atcaggacgc gcaggtagtg tcccaacaac ccccaacaat
     4621 ttttactcat cagatcaata atcgtgtgac ctacggtgga gccatagacg ctcaggacgc
     4681 caaaaccttc gggccccaaa atgaagagaa ttgggaacat accccagctc acgaaaaaca
     4741 gccatgccat gccggtcacg acctggcggc accgaccctt tggcacagta tgataaccct
     4801 cgatatatgc tttggcggcg tgaaaaaatg tgttcgcgcc atagcacaat ccaagacaaa
     4861 agaagatgac tttaacatag ccggttgcca tggcgctggt agccccccac acgatagtcc
     4921 cgatgtctga gacaaggagt cccatggttc tcctgctgta gtcgttgctc aggccggtga
     4981 ggttgctcag gcggataagg atgacaggac aagtgagcag ccaggttgca tagcgcagcc
     5041 actgcacccg gtgtcctgtg gcaaggtaga gcatagaggg attcttaaac tcaaaaaaga
     5101 actcgagaat caccttaacc atttcaatgg cgcacacata gatctcctcc cagccgcatg
     5161 tagatttcca ggtttggtag gcatagaaca tcagcagcaa aatgctgaat cctgctgcaa
     5221 gccactgcag gacatttgac gcggtctgag cgccgttcgt gccgcgagat tcaatccatc
     5281 cggcacagta acattgatcc tcagggacca ggacggaccc gttcaccacc acaggattag
     5341 taacgaacaa aagttcgcgt ccgacggcag acaaagcgcc gccatagtcc atggtggcgg
     5401 cgcgccacaa cttcgtatag aaaggtatat acgaagttgt tccggagtcg actctagagg
     5461 atccggtacc tcacgacacc tgaaatggaa gaaaaaaact ttgaaccact gtctgaggct
     5521 tgagaatgaa ccaagatcca aactcaaaaa gggcaaattc caaggagaat tacatcaagt
     5581 gccaagctgg cctaacttca gtctccaccc actcagtgtg gggaaactcc atcgcataaa
     5641 acccctcccc ccaacctaaa gacgacgtac tccaaaagct cgagaactaa tcgaggtgcc
     5701 tggacggcgc ccggtactcc gtggagtcac atgaagcgac ggctgaggac ggaaaggccc
     5761 ttttcctttg tgtgggtgac tcacccgccc gctctcccga gcgccgcgtc ctccattttg
     5821 agctccctgc agcagggccg ggaagcggcc atctttccgc tcacgcaact ggtgccgacc
     5881 gggccagcct tgccgcccag ggcggggcga tacacggcgg cgcgaggcca ggcaccagag
     5941 caggccggcc agcttgagac tacccccgtc cgattctcgg tggccgcgct cgcaggcccc
     6001 gcctcgccga acatgtgcgc tgggacgcac gggccccgtc gccgcccgcg gccccaaaaa
     6061 ccgaaatacc agtgtgcaga tcttggcccg catttacaag actatcttgc cagaaaaaaa
     6121 gcgtcgcagc aggtcatcaa aaattttaaa tggctagaga cttatcgaaa gcagcgagac
     6181 aggcgcgaag gtgccaccag attcgcacgc ggcggcccca gcgcccaggc caggcctcaa
     6241 ctcaagcacg aggcgaaggg gctccttaag cgcaaggcct cgaactctcc cacccacttc
     6301 caacccgaag ctcgggatca agaatcacgt actgcagcca ggtggaagta attcaaggca
     6361 cgcaagggcc ataacccgta aagaggccag gcccgcggga accacacacg gcacttacct
     6421 gtgttctggc ggcaaacccg ttgcgaaaaa gaacgttcac ggcgactact gcacttatat
     6481 acggttctcc cccaccctcg ggaaaaaggc ggagccagta cacgacatca ctttcccagt
     6541 ttaccccgcg ccaccttctc taggcaccgg ttcaattgcc gacccctccc cccaacttct
     6601 cggggactgt gggcgatgtg cgctctgccc actgacgggc accggagcca attcccactc
     6661 ctttcaagac ctagaaggtc cattagctgc aaagattcct ctctgtttaa aactttatcc
     6721 atctttgcaa agcttacgcg tgcggccgca ggaaccccta gtgatggagt tggccactcc
     6781 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     6841 cggcctcagt gagcgagcga gcgcgcagct gcctgcagga catgtgagca aaaggccagc
     6901 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6961 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7021 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7081 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7141 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7201 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7261 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7321 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7381 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7441 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7501 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7561 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7621 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7681 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7741 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacg
//
