LOCUS       sequence-181395-        7029 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  produces AAV expressing Cal-light 
            TM-CaM-NES-TEV-N-AsLOV2-TEVseq-tTA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7029)
  AUTHORS   Lee D, Hyun JH, Jung K, Hannan P, Kwon HB
  TITLE     A calcium- and light-gated switch to induce gene expression in 
            activated neurons.
  JOURNAL   Nat Biotechnol. 2017 Jun 26. doi: 10.1038/nbt.3902.
  PUBMED    28650460
REFERENCE   2  (bases 1 to 7029)
  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: "Nat 
            Biotechnol. 2017 Jun 26. doi: 10.1038/nbt.3902."
COMMENT     Sequence Label: pAAV-TM-CaM-NES-TEV-N-AsLOV2-TEVseq-tTA
FEATURES             Location/Qualifiers
     source          1..7029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..453
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     promoter        474..492
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     474..491
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     sig_peptide     511..570
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             574..603
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             607..753
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             1294..1320
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2113..2133
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(2134..2751,2752..3138)
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="SRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVK
                     NKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRP
                     TEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTD
                     SMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTIE
                     GLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHAD
                     ALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(2204..2223)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     misc_feature    3157..3745
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3210..3230)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3628..3639)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3786..3993
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4163..4292
                     /label=AAV2 ITR
     rep_origin      complement(4529..5117)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4609..4628)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5288..6079,6080..6148))
                     /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     5911..5930
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6149..6253)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6279..6734)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6416..6437)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6628..6647
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6806..6935
                     /label=AAV2 ITR
ORIGIN
        1 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
       61 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      121 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      181 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      241 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      301 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      361 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      421 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagag gatctaagct ttcatttagg
      481 tgacactata gaatacaagc tacttgttct atggagacag acacactcct gctatgggta
      541 ctgctgctct gggttccagg ttccactggt gacgaacaaa aactcatctc agaagaggat
      601 ctgaatgctg tgggccagga cacgcaggag gtcatcgtgg tgccacactc cttgcccttt
      661 aaggtggtgg tgatctcagc catcctggcc ctggtggtgc tcaccatcat ctcccttatc
      721 atcctcatca tgctttggca gaagaagcca cgtggtggct ctggaggtct cgagatgccg
      781 gaccaactga ctgaagagca gatcgcagaa tttaaagagg ctttctccct atttgacaag
      841 gacggggatg ggacaataac aaccaaggag ctggggacgg tgatgcggtc tctggggcag
      901 aaccccacag aagcagagct gcaagacatg atcaatgaag tagatgccga cggtgacggc
      961 acaatcgact tccctgagtt cctgacaatg atggcaagaa aaatgaaata cagggacacg
     1021 gaagaagaaa ttagagaagc gttcggtgtg tttgataagg atggcaatgg ctacatcagt
     1081 gcagcagagc ttcgccacgt gatgacaaac cttggagaga agttaacaga tgaagaggtt
     1141 gatgaaatga tcagggaagc agacatcgat ggggatggtc aggtaaacta cgaagagttt
     1201 gtacaaatga tgacagcgaa ggcatgcggt ggcggtggct ctggaggtgg tgggtccgga
     1261 ggaggcggcc gctccggtgg atccatgctt caacttcctc ctcttgaacg tcttactctc
     1321 gagatgggcg agagcctgtt caagggacct agggactaca accctatcag tagcacaatt
     1381 tgtcacctga ccaacgagag tgatggccac acaacaagcc tgtacggcat cgggttcgga
     1441 ccctttatca tcaccaacaa gcacctgttc aggcggaata atggcactct gctggtgcag
     1501 agcctgcacg gggtgttcaa agtgaagaac acaaccactc tgcagcagca cctgatcgat
     1561 gggcgggata tgatcatcat taggatgccc aaggacttcc ccccttttcc tcagaaactg
     1621 aagttccgag agccccagag agaggagaga atctgtctgg tgaccacaaa ctttcaggag
     1681 ctcggtagtg gtagtgggga gtttctggca accacactgg aacggatcga gaaaaatttc
     1741 gtgattactg atccgagact gcctgacaac ccaatcattt ttgcgagcga ttccttcctg
     1801 cagctgacag aatattctcg ggaagagatc ctggggcgca attgccgttt tctgcaggga
     1861 cccgagacag accgtgccac tgttcggaaa atcagagatg ctattgacaa ccagactgaa
     1921 gtgaccgttc agctgatcaa ttataccaag agcggcaaga agttctggaa cgtgttccac
     1981 ctgcagccga tgcgcgatta taagggcgac gtccagtact tcattggcgt gcagctggat
     2041 ggcaccgaac gtcttcatgg cgccgctgag cgtgaggcgg tctgcctgat caaaaagaca
     2101 gcctttcaga ttgagaacct gtacttccag ggctctagat tagataaaag taaagtgatt
     2161 aacagcgcat tagagctgct taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc
     2221 gcccagaagc taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa taagcgggct
     2281 ttgctcgacg ccttagccat tgagatgtta gataggcacc atactcactt ttgcccttta
     2341 gaaggggaaa gctggcaaga ttttttacgt aataacgcta aaagttttag atgtgcttta
     2401 ctaagtcatc gcgatggagc aaaagtacat ttaggtacac ggcctacaga aaaacagtat
     2461 gaaactctcg aaaatcaatt agccttttta tgccaacaag gtttttcact agagaatgca
     2521 ttatatgcac tcagcgctgt ggggcatttt actttaggtt gcgtattgga agatcaagag
     2581 catcaagtcg ctaaagaaga aagggaaaca cctactactg atagtatgcc gccattatta
     2641 cgacaagcta tcgaattatt tgatcaccaa ggtgcagagc cagccttctt attcggcctt
     2701 gaattgatca tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc cgcgtacagc
     2761 cgcgcgcgta cgaaaaacaa ttacgggtct accatcgagg gcctgctcga tctcccggac
     2821 gacgacgccc ccgaagaggc ggggctggcg gctccgcgcc tgtcctttct ccccgcggga
     2881 cacacgcgca gactgtcgac ggcccccccg accgatgtca gcctggggga cgagctccac
     2941 ttagacggcg aggacgtggc gatggcgcat gccgacgcgc tagacgattt cgatctggac
     3001 atgttggggg acggggattc cccgggtccg ggatttaccc cccacgactc cgccccctac
     3061 ggcgctctgg atatggccga cttcgagttt gagcagatgt ttaccgatgc ccttggaatt
     3121 gacgagtacg gtgggtaggc tagccagctt atcgataatc aacctctgga ttacaaaatt
     3181 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3241 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3301 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3361 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3421 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3481 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3541 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     3601 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3661 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3721 atctcccttt gggccgcctc cccgcatcga taccgtcgac ccgggcggcc gctcataaag
     3781 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3841 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3901 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3961 ggattgggaa gacaatagca ggcatgctgg ggatgcggtc cggactgtac aattgcattc
     4021 attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct
     4081 acaaatgtgg taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat
     4141 ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg
     4201 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     4261 cgggcggcct cagtgagcga gcgagcgcgc agctgcatta atgaatcggc caacgcgcgg
     4321 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4381 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4441 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4501 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4561 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4621 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4681 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4741 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4801 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4861 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4921 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4981 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5041 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5101 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5161 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5221 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5281 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5341 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5401 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5461 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5521 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5581 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5641 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5701 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5761 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5821 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5881 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5941 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6001 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6061 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6121 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6181 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6241 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     6301 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     6361 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     6421 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     6481 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     6541 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     6601 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     6661 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     6721 gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     6781 gcgggcctct tcgctattac gccagctgcg cgctcgctcg ctcactgagg ccgcccgggc
     6841 aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
     6901 agagggagtg gccaactcca tcactagggg ttccttgtag ttaatgatta acccgccatg
     6961 ctacttatct acgtagccat gctctaggaa gatcgtaggt taattaatct agactgcaga
     7021 gggccctgc
//
