LOCUS       sequence-91176-e        6758 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AAV expression of Mac-GFP under the CaMKII promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6758)
  AUTHORS   Chow BY, Han X, Dobry AS, Qian X, Chuong AS, Li M, Henninger MA, 
            Belfort GM, Lin Y, Monahan PE, Boyden ES
  TITLE     High-performance genetically targetable optical neural silencing by 
            light-driven proton pumps.
  JOURNAL   Nature. 2010 Jan 7. 463(7277):98-102.
  PUBMED    20054397
REFERENCE   2  (bases 1 to 6758)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2010 Jan 7. 463(7277):98-102."
COMMENT     Sequence Label: pAAV-CaMKII-Mac-GFP
FEATURES             Location/Qualifiers
     source          1..6758
                     /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"
     CDS             join(4919..4921,4922..5635)
                     /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(4919..4921,4922..5632)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4961..4982)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5222..5241)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5569..5590
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5660..6248
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5713..5733)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6131..6142)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6291..6412
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6302..6321)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6356..6375
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   6574..6703
                     /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 caccatgatc gtggaccagt tcgaggaggt gctgatgaag accagccagc tgttcccact
     4021 gccaaccgct acccagagcg cccagccaac ccacgtggcc cccgtgccaa ccgtgctgcc
     4081 cgacaccccc atctacgaga ccgtgggcga cagcggcagc aagaccctgt gggtggtgtt
     4141 cgtgctgatg ctgatcgcca gcgccgcctt caccgccctg agctggaaga tccccgtgaa
     4201 caggaggctg taccacgtga tcaccaccat catcaccctg accgccgccc tgagctactt
     4261 cgctatggct accggccacg gagtggccct gaacaagatc gtgatcagga cccagcacga
     4321 ccacgtgccc gacacctacg agaccgtgta ccgacaggtg tactacgcca ggtacatcga
     4381 ctgggctatc accaccccac tgctgctgct ggacctggga ctgctggctg gaatgagcgg
     4441 agcccacatc ttcatggcca tcgtggctga cctgatcatg gtgctgaccg gcctgttcgc
     4501 tgctttcggc agcgagggaa ccccacagaa gtggggatgg tacaccatcg cctgcatcgc
     4561 ctacatcttc gtggtgtggc acctggtgct gaacggcggc gccaacgcca gggtgaaggg
     4621 cgagaagctg aggagcttct tcgtggccat cggagcttac accctgatcc tgtggaccgc
     4681 ttacccaatc gtgtggggac tggctgacgg agctaggaag atcggagtgg acggagagat
     4741 catcgcttac gctgtgctgg acgtgctggc taagggagtg ttcggagctt ggctgctggt
     4801 gacccacgcc aacctgaggg agagcgacgt ggagctgaac ggcttctggg ccaacggcct
     4861 gaacagggag ggcgccatca ggatcggcga ggacgacggc gcccgaccgg tagtagcagt
     4921 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     4981 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     5041 gctgaccctg aagttcattt gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     5101 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     5161 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     5221 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     5281 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     5341 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     5401 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     5461 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     5521 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     5581 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaagaatt
     5641 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     5701 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     5761 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     5821 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     5881 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     5941 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     6001 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     6061 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     6121 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     6181 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     6241 ctccccgcat cgataccgtc gacccgggcg gccgcttcga gcagacatga taagatacat
     6301 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     6361 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     6421 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa
     6481 gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt cctagagcat ggctacgtag
     6541 ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca
     6601 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     6661 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcatt aatgaatcgg
     6721 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgct
//
