LOCUS       sequence-343524-        5616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of InteinC-tTAC in CaMKII positive cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5616)
  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 5616)
  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-tTAc
FEATURES             Location/Qualifiers
     source          1..5616
                     /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"
     CDS             1605..1715
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     misc_feature    1743..2331
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1796..1816)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2214..2225)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2363..2839
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2562..2581)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2879..3019
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2879..3008
                     /label=AAV2 ITR
     rep_origin      3094..3549
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3181..3200)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3391..3412
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3566..3585)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3685..3707
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3745..3763)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3831..3935
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3936..4004,4005..4796)
                     /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(4154..4173)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4967..5555
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5456..5475
                     /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 tgatgctgaa gaagatcctg aagatcgagg
     1501 agctggacga gcgcgagctg atcgacatcg aggtgagcgg caaccacctg ttctacgcca
     1561 acgacatcct gacccacaac agcagcagca gcgacgtgcc ggccgacgcc cttgacgatt
     1621 ttgacttaga catgctccca gccgatgccc ttgacgactt tgaccttgat atgctgcctg
     1681 ctgacgctct tgacgatttt gaccttgaca tgctccccgg gtaagatatc aagcttatcg
     1741 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     1801 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     1861 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     1921 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     1981 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     2041 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     2101 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     2161 tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     2221 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     2281 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc
     2341 gagcgctgct cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct
     2401 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat
     2461 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
     2521 caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg
     2581 gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct
     2641 cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt
     2701 tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta
     2761 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
     2821 tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag
     2881 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     2941 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     3001 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3061 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     3121 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     3181 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3241 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3301 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3361 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3421 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     3481 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     3541 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     3601 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     3661 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     3721 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     3781 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3841 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3901 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3961 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4021 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4081 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4141 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4201 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4261 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4321 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4381 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4441 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4501 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4561 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4621 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4681 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4741 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4801 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4861 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4921 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4981 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5041 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5101 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5161 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5221 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5281 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5341 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5401 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5461 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5521 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5581 ggttcctggc cttttgctgg ccttttgctc acatgt
//
