LOCUS       sequence-337521-        6956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV Soma-targeted Cal-Light with Kv2.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6956)
  AUTHORS   Hyun JH, Nagahama K, Namkung H, Mignocchi N, Roh SE, Hannan P, 
            Krussel S, Kwak C, McElroy A, Liu B, Cui M, Lee S, Lee D, Huganir 
            RL, Worley PF, Sawa A, Kwon HB
  TITLE     Tagging active neurons by soma-targeted Cal-Light.
  JOURNAL   Nat Commun. 2022 Dec 12;13(1):7692. doi: 10.1038/s41467-022-35406-y.
  PUBMED    36509775
REFERENCE   2  (bases 1 to 6956)
  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 
            Commun."; date: "2022-12-12"; volume: "13(1):7692. doi"; pages: " 
            10.1038/s41467-022-35406-y"
COMMENT     Sequence Label: pAAV-pCMV-TM-Kv2.1-CaM-NES-TevN-AsLOV2-TEVseq-tTA
FEATURES             Location/Qualifiers
     source          1..6956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..126
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        159..1143
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1179..1197
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1179..1196
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     sig_peptide     1216..1275
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             1279..1308
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1312..1458
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             2179..2205
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2998..3018
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(3019..3636,3637..4023)
                     /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(3089..3108)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(4032..4051)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4033..4051)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4056..4190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4080..4099)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4134..4153
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4219..4359
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4219..4348
                     /label=AAV2 ITR
     rep_origin      4434..4889
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4521..4540)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4731..4752
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4906..4925)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5025..5047
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5085..5103)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5171..5275
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5276..5344,5345..6136)
                     /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(5494..5513)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6307..6895
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6796..6815
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
      121 gttcctgcgg ccgcacgcgt ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg
      181 cgtatatgga ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg
      241 cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg
      301 cggacttggc accatgccaa ttcaatatgg cggacctggc actgtgccaa ctggggaggg
      361 gtctacttgg cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat
      421 attgaattgg catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata
      481 taggcaatat ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt
      541 attggcccta tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc
      601 tcccaatggg cggtcccata taccatatat ggggcttcct aataccgccc atagccactc
      661 ccccattgac gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc
      721 ctattgacgt atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc
      781 tcaatgccca ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg
      841 cccattcata tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg
      901 gcagtacatc aatatctatt aatagtaact tggcaagtac attactattg gaagtacgcc
      961 agggtacatt ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc
     1021 caagtacatc cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg
     1081 acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg
     1141 aaccgccatt ctgcctgggg acgtcggagc aagctttcat ttaggtgaca ctatagaata
     1201 caagctactt gttctatgga gacagacaca ctcctgctat gggtactgct gctctgggtt
     1261 ccaggttcca ctggtgacga acaaaaactc atctcagaag aggatctgaa tgctgtgggc
     1321 caggacacgc aggaggtcat cgtggtgcca cactccttgc cctttaaggt ggtggtgatc
     1381 tcagccatcc tggccctggt ggtgctcacc atcatctccc ttatcatcct catcatgctt
     1441 tggcagaaga agccacgtca gtcccagcct attctgaaca ccaaagagat ggccccccag
     1501 agtaaacccc ctgaggagct ggaaatgtcc tcaatgccct cccctgtggc acctcttcct
     1561 gcaagaacag agggcgtgat cgacatgagg tccatgagtt ctattgactc ttttatctca
     1621 tgcgccaccg attttccaga ggctacgcgg tttctcgaga tgccggacca actgactgaa
     1681 gagcagatcg cagaatttaa agaggctttc tccctatttg acaaggacgg ggatgggaca
     1741 ataacaacca aggagctggg gacggtgatg cggtctctgg ggcagaaccc cacagaagca
     1801 gagctgcaag acatgatcaa tgaagtagat gccgacggtg acggcacaat cgacttccct
     1861 gagttcctga caatgatggc aagaaaaatg aaatacaggg acacggaaga agaaattaga
     1921 gaagcgttcg gtgtgtttga taaggatggc aatggctaca tcagtgcagc agagcttcgc
     1981 cacgtgatga caaaccttgg agagaagtta acagatgaag aggttgatga aatgatcagg
     2041 gaagcagaca tcgatgggga tggtcaggta aactacgaag agtttgtaca aatgatgaca
     2101 gcgaaggcat gcggtggcgg tggctctgga ggtggtgggt ccggaggagg cggccgctcc
     2161 ggtggatcca tgcttcaact tcctcctctt gaacgtctta ctctcgaaat gggcgagagc
     2221 ctgttcaagg gacctaggga ctacaaccct atcagtagca caatttgtca cctgaccaac
     2281 gagagtgatg gccacacaac aagcctgtac ggcatcgggt tcggaccctt tatcatcacc
     2341 aacaagcacc tgttcaggcg gaataatggc actctgctgg tgcagagcct gcacggggtg
     2401 ttcaaagtga agaacacaac cactctgcag cagcacctga tcgatgggcg ggatatgatc
     2461 atcattagga tgcccaagga cttcccccct tttcctcaga aactgaagtt ccgagagccc
     2521 cagagagagg agagaatctg tctggtgacc acaaactttc aggagctcgg tagtggtagt
     2581 ggggagtttc tggcaaccac actggaacgg atcgagaaaa atttcgtgat tactgatccg
     2641 agactgcctg acaacccaat catttttgcg agcgattcct tcctgcagct gacagaatat
     2701 tctcgggaag agatcctggg gcgcaattgc cgttttctgc agggacccga gacagaccgt
     2761 gccactgttc ggaaaatcag agatgctatt gacaaccaga ctgaagtgac cgttcagctg
     2821 atcaattata ccaagagcgg caagaagttc tggaacgtgt tccacctgca gccgatgcgc
     2881 gattataagg gcgacgtcca gtacttcatt ggcgtgcagc tggatggcac cgaacgtctt
     2941 catggcgccg ctgagcgtga ggcggtctgc ctgatcaaaa agacagcctt tcagattgag
     3001 aacctgtact tccagggctc tagattagat aaaagtaaag tgattaacag cgcattagag
     3061 ctgcttaatg aggtcggaat cgaaggttta acaacccgta aactcgccca gaagctaggt
     3121 gtagagcagc ctacattgta ttggcatgta aaaaataagc gggctttgct cgacgcctta
     3181 gccattgaga tgttagatag gcaccatact cacttttgcc ctttagaagg ggaaagctgg
     3241 caagattttt tacgtaataa cgctaaaagt tttagatgtg ctttactaag tcatcgcgat
     3301 ggagcaaaag tacatttagg tacacggcct acagaaaaac agtatgaaac tctcgaaaat
     3361 caattagcct ttttatgcca acaaggtttt tcactagaga atgcattata tgcactcagc
     3421 gctgtggggc attttacttt aggttgcgta ttggaagatc aagagcatca agtcgctaaa
     3481 gaagaaaggg aaacacctac tactgatagt atgccgccat tattacgaca agctatcgaa
     3541 ttatttgatc accaaggtgc agagccagcc ttcttattcg gccttgaatt gatcatatgc
     3601 ggattagaaa aacaacttaa atgtgaaagt gggtccgcgt acagccgcgc gcgtacgaaa
     3661 aacaattacg ggtctaccat cgagggcctg ctcgatctcc cggacgacga cgcccccgaa
     3721 gaggcggggc tggcggctcc gcgcctgtcc tttctccccg cgggacacac gcgcagactg
     3781 tcgacggccc ccccgaccga tgtcagcctg ggggacgagc tccacttaga cggcgaggac
     3841 gtggcgatgg cgcatgccga cgcgctagac gatttcgatc tggacatgtt gggggacggg
     3901 gattccccgg gtccgggatt taccccccac gactccgccc cctacggcgc tctggatatg
     3961 gccgacttcg agtttgagca gatgtttacc gatgcccttg gaattgacga gtacggtggg
     4021 taggctagcc tccctatagt gagtcgtatt acgtagatcc agacatgata agatacattg
     4081 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     4141 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt ggtaaccacg
     4201 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     4261 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     4321 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     4381 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     4441 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4501 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4561 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4621 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     4681 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4741 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     4801 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4861 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     4921 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4981 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5041 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     5101 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     5161 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5221 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5281 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5341 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5401 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5461 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5521 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5581 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5641 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5701 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5761 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5821 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5881 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5941 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6001 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6061 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6121 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6181 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6241 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6301 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6361 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6421 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6481 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6541 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6601 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6661 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6721 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6781 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6841 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6901 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
