LOCUS       sequence-91102-e        6797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of Chronos-GFP under the CaMKII promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6797)
  AUTHORS   Klapoetke NC, Murata Y, Kim SS, Pulver SR, Birdsey-Benson A, Cho YK,
            Morimoto TK, Chuong AS, Carpenter EJ, Tian Z, Wang J, Xie Y, Yan Z, 
            Zhang Y, Chow BY, Surek B, Melkonian M, Jayaraman V, 
            Constantine-Paton M, Wong GK, Boyden ES
  TITLE     Independent optical excitation of distinct neural populations.
  JOURNAL   Nat Methods. 2014 Mar;11(3):338-46. doi: 10.1038/nmeth.2836. Epub 
            2014 Feb 9.
  PUBMED    24509633
REFERENCE   2  (bases 1 to 6797)
  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"; doi: 
            "10.1038/nmeth.2836"; journalName: "Nat Methods"; date: "2014-03"; 
            volume: "11"; issue: "3"; pages: "338-46"
COMMENT     Sequence Label: pAAV-CaMKII-Chronos-GFP
FEATURES             Location/Qualifiers
     source          1..6797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2459..2588
                     /label=AAV2 ITR
     promoter        2661..3949
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      3959..3968
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4958..4960,4961..5674)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4958..4960,4961..5671)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5000..5021)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5261..5280)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5608..5629
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5699..6287
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5752..5772)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6170..6181)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6330..6451
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6341..6360)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6395..6414
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   6613..6742
                     /label=AAV2 ITR
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagatcgta ggttaattaa cattatggcc ttaggtcact tcatctccat ggggttcttc
     2701 ttctgatttt ctagaaaatg agatgggggt gcagagagct tcctcagtga cctgcccagg
     2761 gtcacatcag aaatgtcaga gctagaactt gaactcagat tactaatctt aaattccatg
     2821 ccttgggggc atgcaagtac gatatacaga aggagtgaac tcattagggc agatgaccaa
     2881 tgagtttagg aaagaagagt ccagggcagg gtacatctac accacccgcc cagccctggg
     2941 tgagtccagc cacgttcacc tcattatagt tgcctctctc cagtcctacc ttgacgggaa
     3001 gcacaagcag aaactgggac aggagcccca ggagaccaaa tcttcatggt ccctctggga
     3061 ggatgggtgg ggagagctgt ggcagaggcc tcaggagggg ccctgctgct cagtggtgac
     3121 agataggggt gagaaagcag acagagtcat tccgtcagca ttctgggtct gtttggtact
     3181 tcttctcacg ctaaggtggc ggtgtgatat gcacaatggc taaaaagcag ggagagctgg
     3241 aaagaaacaa ggacagagac agaggccaag tcaaccagac caattcccag aggaagcaaa
     3301 gaaaccatta cagagactac aagggggaag ggaaggagag atgaattagc ttcccctgta
     3361 aaccttagaa cccagctgtt gccagggcaa cggggcaata cctgtctctt cagaggagat
     3421 gaagttgcca gggtaactac atcctgtctt tctcaaggac catcccagaa tgtggcaccc
     3481 actagccgtt accatagcaa ctgcctcttt gccccactta atcccatccc gtctgttaaa
     3541 agggccctat agttggaggt gggggaggta ggaagagcga tgatcacttg tggactaagt
     3601 ttgttcgcat ccccttctcc aaccccctca gtacatcacc ctgggggaac agggtccact
     3661 tgctcctggg cccacacagt cctgcagtat tgtgtatata aggccagggc aaagaggagc
     3721 aggttttaaa gtgaaaggca ggcaggtgtt ggggaggcag ttaccggggc aacgggaaca
     3781 gggcgtttcg gaggtggttg ccatggggac ctggatgctg acgaaggctc gcgaggctgt
     3841 gagcagccac agtgccctgc tcagaagccc caagctcgtc agtcaagccg gttctccgtt
     3901 tgcactcagg agcacgggca ggcgagtggc ccctagttct gggggcagcg ggggatccgc
     3961 caccatggaa acagccgcca caatgaccca cgcctttatc tcagccgtgc ctagcgccga
     4021 agccacaatt agaggcctgc tgagcgccgc agcagtggtg acaccagcag cagacgctca
     4081 cggagaaacc tctaacgcca caacagccgg agccgatcac ggttgcttcc cccacatcaa
     4141 ccacggaacc gagctgcagc acaagatcgc agtgggactc cagtggttca ccgtgatcgt
     4201 ggctatcgtg cagctcatct tctacggttg gcacagcttc aaggccacaa ccggctggga
     4261 ggaggtctac gtctgcgtga tcgagctcgt caagtgcttc atcgagctgt tccacgaggt
     4321 cgacagccca gccacagtgt accagaccaa cggaggagcc gtgatttggc tgcggtacag
     4381 catgtggctc ctgacttgcc ccgtgatcct gatccacctg agcaacctga ccggactgca
     4441 cgaagagtac agcaagcgga ccatgaccat cctggtgacc gacatcggca acatcgtgtg
     4501 ggggatcaca gccgccttta caaagggccc cctgaagatc ctgttcttca tgatcggcct
     4561 gttctacggc gtgacttgct tcttccagat cgccaaggtg tatatcgaga gctaccacac
     4621 cctgcccaaa ggcgtctgcc ggaagatttg caagatcatg gcctacgtct tcttctgctc
     4681 ttggctgatg ttccccgtga tgttcatcgc cggacacgag ggactgggcc tgatcacacc
     4741 ttacaccagc ggaatcggcc acctgatcct ggatctgatc agcaagaaca cttggggctt
     4801 cctgggccac cacctgagag tgaagatcca cgagcacatc ctgatccacg gcgacatccg
     4861 gaagacaacc accatcaacg tggccggcga gaacatggag atcgagacct tcgtcgacga
     4921 ggaggaggag ggaggagtgg cggcaccggt agtagcagtg agcaagggcg aggagctgtt
     4981 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     5041 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatttg
     5101 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     5161 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     5221 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     5281 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     5341 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     5401 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     5461 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     5521 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     5581 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     5641 gatcactctc ggcatggacg agctgtacaa gtaagaattc gatatcaagc ttatcgataa
     5701 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     5761 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     5821 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     5881 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     5941 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     6001 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     6061 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     6121 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     6181 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     6241 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg
     6301 acccgggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     6361 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     6421 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     6481 tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     6541 taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa
     6601 tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     6661 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     6721 cagtgagcga gcgagcgcgc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6781 tttgcgtatt gggcgct
//
