LOCUS       sequence-343530-        7041 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of EBFP and tTA-InteinN in CaMKII positive cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7041)
  AUTHORS   Chen HS, Zhang XL, Yang RR, Wang GL, Zhu XY, Xu YF, Wang DY, Zhang 
            N, Qiu S, Zhan LJ, Shen ZM, Xu XH, Long G, Xu C
  TITLE     An intein-split transactivator for intersectional neural imaging and
            optogenetic manipulation.
  JOURNAL   Nat Commun. 2022 Jun 23;13(1):3605. doi: 10.1038/s41467-022-31255-x.
  PUBMED    35739125
REFERENCE   2  (bases 1 to 7041)
  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: "2022-06-23"; volume: "13(1):3605. doi"; pages: " 
            10.1038/s41467-022-31255-x"
COMMENT     Sequence Label: pAAV-pCaMKII-EBFP-tTAn
FEATURES             Location/Qualifiers
     source          1..7041
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        153..1442
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1464..1473
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1470..1472,1473..1475,1476..2186)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(1515..1536)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2196..2249
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    3168..3756
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3221..3241)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3639..3650)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3788..4264
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3987..4006)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4304..4444
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4304..4433
                     /label=AAV2 ITR
     rep_origin      4519..4974
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4606..4625)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4816..4837
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4991..5010)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5110..5132
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5170..5188)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5256..5360
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5361..5429,5430..6221)
                     /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(5579..5598)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6392..6980
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6881..6900
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtttaatt aacattatgg ccttaggtca cttcatctcc
      181 atggggttct tcttctgatt ttctagaaaa tgagatgggg gtgcagagag cttcctcagt
      241 gacctgccca gggtcacatc agaaatgtca gagctagaac ttgaactcag attactaatc
      301 ttaaattcca tgccttgggg gcatgcaagt acgatataca gaaggagtga actcattagg
      361 gcagatgacc aatgagttta ggaaagaaga gtccagggca gggtacatct acaccacccg
      421 cccagccctg ggtgagtcca gccacgttca cctcattata gttgcctctc tccagtccta
      481 ccttgacggg aagcacaagc agaaactggg acaggagccc caggagacca aatcttcatg
      541 gtccctctgg gaggatgggt ggggagagct gtggcagagg cctcaggagg ggccctgctg
      601 ctcagtggtg acagataggg gtgagaaagc agacagagtc attccgtcag cattctgggt
      661 ctgtttggta cttcttctca cgctaaggtg gcggtgtgat atgcacaatg gctaaaaagc
      721 agggagagct ggaaagaaac aaggacagag acagaggcca agtcaaccag accaattccc
      781 agaggaagca aagaaaccat tacagagact acaaggggga agggaaggag agatgaatta
      841 gcttcccctg taaaccttag aacccagctg ttgccagggc aacggggcaa tacctgtctc
      901 ttcagaggag atgaagttgc cagggtaact acatcctgtc tttctcaagg accatcccag
      961 aatgtggcac ccactagccg ttaccatagc aactgcctct ttgccccact taatcccatc
     1021 ccgtctgtta aaagggccct atagttggag gtgggggagg taggaagagc gatgatcact
     1081 tgtggactaa gtttgttcgc atccccttct ccaaccccct cagtacatca ccctggggga
     1141 acagggtcca cttgctcctg ggcccacaca gtcctgcagt attgtgtata taaggccagg
     1201 gcaaaggagg agcaggtttt aaagtgaaag gcaggcaggt gttggggagg cagttaccgg
     1261 ggcaacggga acagggcgtt tcggaggtgg ttgccatggg gacctggatg ctgacgaagg
     1321 ctcgcgaggc tgtgagcagc cacagtgccc tgctcagaag ccccaagctc gtcagtcaag
     1381 ccggttctcc gtttgcactc aggagcacgg gcaggcgagt ggcccctagt tctgggggca
     1441 gcgggggatc cccgggtacc ggtgccacca tggtgagcaa gggcgaggag ctgttcaccg
     1501 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtga
     1561 ggggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc atctgcacca
     1621 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgagccac ggcgtgcagt
     1681 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1741 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcacctac aagacccgcg
     1801 ccgaggtgaa gttcgagggc gacaccctag tgaaccgcat cgagctgaag ggcgtcgact
     1861 tcaaggagga cggcaacatc ctggggcaca agctggagta caacttcaac agccacaaca
     1921 tctatatcat ggccgtcaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     1981 acgtggagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     2041 acggccccgt gctgctgccc gacagccact acctgagcac ccagtccgtg ctgagcaaag
     2101 accccaacga gaagcgcgat cacatggtcc tgctggagtt ccgcaccgcc gccgggatca
     2161 ctctcggcat ggacgagctg tacaagggca gtggagaggg cagaggaagt ctgctaacat
     2221 gcggtgacgt cgaggagaat cctggcccat ctagactgga caagagcaaa gtcataaact
     2281 ctgctctgga attactcaat gaagtcggta tcgaaggcct gacgacaagg aaactcgctc
     2341 aaaagctggg agttgagcag cctaccctgt actggcacgt gaagaacaag cgggccctgc
     2401 tcgatgccct ggcaatcgag atgctggaca ggcatcatac ccacttctgc cccctggaag
     2461 gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc attccgctgt gctctcctct
     2521 cacatcgcga cggggctaaa gtgcatctcg gcacccgccc aacagagaaa cagtacgaaa
     2581 ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt ctccctggag aacgcactgt
     2641 acgctctgtc cgccgtgggc cactttacac tgggctgcgt attggaggat caggagcatc
     2701 aagtagcaaa agaggaaaga gagacaccta ccaccgattc tatgccccca cttctgagac
     2761 aagcaattga gctgttcgac catcagggag ccgaacctgc cttccttttc ggcctggaac
     2821 taatcatatg tggcctggag aaacagctaa agtgcgaaag cggcgggacc cgcagcggct
     2881 actgcctgga cctgaagacc caggtgcaga ccccccaggg catgaaggag atcagcaaca
     2941 tccaggtggg cgacctggtg ctgagcaaca ccggctacaa cgaggtgctg aacgtgttcc
     3001 ccaagagcaa gaagaagagc tacaagatca ccctggagga cggcaaggag atcatctgca
     3061 gcgaggagca cctgttcccc acccagaccg gcgagatgaa catcagcggc ggcctgaagg
     3121 agggcatgtg cctgtacgtg aaggagtaag atatcaagct tatcgataat caacctctgg
     3181 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3241 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     3301 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     3361 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     3421 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     3481 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     3541 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca
     3601 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     3661 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     3721 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga
     3781 gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc
     3841 cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta
     3901 ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg
     3961 gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca
     4021 atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc
     4081 cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta
     4141 gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta
     4201 cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt
     4261 ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     4321 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     4381 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     4441 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4501 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4561 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     4621 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4681 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     4741 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4801 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     4861 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     4921 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     4981 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     5041 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     5101 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     5161 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     5221 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5281 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5341 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     5401 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5461 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5521 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5581 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5641 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5701 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5761 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5821 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5881 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5941 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6001 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6061 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6121 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6181 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     6241 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6301 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6361 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6421 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6481 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6541 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6601 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6661 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6721 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6781 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6841 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6901 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6961 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7021 gctggccttt tgctcacatg t
//
