LOCUS       AAV_CaMKIIa_ChRg        6945 bp ds-DNA     circular     22-JAN-2019
DEFINITION  Forebrain principle neuron specific expression of GCaMP6f Ca sensor
            and physically separate nuclear localized dTomato fluorophore .
ACCESSION   .
VERSION     .
KEYWORDS    AAV-CaMKIIa-GCaMP6f-P2A-nls-dTomato
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6945)
  TITLE     Ting GCaMP6 plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6945)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, Aug 12, 2018 from SnapGene 3.3.4
            http://www.snapgene.com
COMMENT     
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     repeat_region   1..141
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
                     /label=AAV2 ITR
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     repeat_region   12..141
                     /label=AAV2 ITR(1)
                     /ApEinfo_label=AAV2 ITR
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             1468..2550
                     /codon_start=1
                     /translation="MSRRPWLLALALAVALAAGSAGAAHIVMVDAYKPTKSTGSDATV
                     P
                     VATQDGPDYVFHRAHERMLFQTSYTLENNGSVICIPNNGQCFCLAWLKSNGTNAEKLA
                     A
                     NILQWITFALSALCLMFYGYQTWKSTCGWEEIYVATIEMIKFIIEYFHEFDSPAMLFL
                     Y
                     GGNITPWLRYAEWLLTCPVILIHLSNITGLSEAYNKRTMALLVSDLGTICMGVTAALA
                     T
                     GWVKWLFYCIGLVYGTQTFYNAGIIYVEAYHTVPKGRCRQVVTGMAWLFFVSWGMFPI
                     L
                     FILGPEGFGVLSVAGSTIGHTIADLLSKNIWGLLGHFLRIKIHEHILIHGDIRKTTKL
                     N IGGTEIEVETLVEDEAEAGAV"
                     /label=ChRger3
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    2560..2619
                     /note="ApEinfo_fwdcolor=#00ff00
                     ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0 ApEinfo_revcolor=green"
                     /label=TS
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             2620..3336
                     /codon_start=1
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     K
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     G
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     V
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     E FVTAAGITLGMDELYK"
                     /label=EYFP (no start)
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    3364..3952
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
                     /label=WPRE
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             complement(3835..3846)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
                     /label=Factor Xa site
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     polyA_signal    3985..4096
                     /note="bovine growth hormone polyadenylation signal"
                     /label=bGH poly(A) signal
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     repeat_region   4208..4348
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
                     /label=AAV2 ITR(2)
                     /ApEinfo_label=AAV2 ITR
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     repeat_region   4208..4337
                     /label=AAV2 ITR(3)
                     /ApEinfo_label=AAV2 ITR
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     rep_origin      4423..4878
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label=f1 ori
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        5160..5264
                     /gene="bla"
                     /label=AmpR promoter
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             5265..6125
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     I
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     S
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     W
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     A
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     S LIKHW"
                     /label=AmpR
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     rep_origin      6296..6884
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label=ori
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtttaat taacattatg gccttaggtc
      181 acttcatctc catggggttc ttcttctgat tttctagaaa atgagatggg ggtgcagaga
      241 gcttcctcag tgacctgccc agggtcacat cagaaatgtc agagctagaa cttgaactca
      301 gattactaat cttaaattcc atgccttggg ggcatgcaag tacgatatac agaaggagtg
      361 aactcattag ggcagatgac caatgagttt aggaaagaag agtccagggc agggtacatc
      421 tacaccaccc gcccagccct gggtgagtcc agccacgttc acctcattat agttgcctct
      481 ctccagtcct accttgacgg gaagcacaag cagaaactgg gacaggagcc ccaggagacc
      541 aaatcttcat ggtccctctg ggaggatggg tggggagagc tgtggcagag gcctcaggag
      601 gggccctgct gctcagtggt gacagatagg ggtgagaaag cagacagagt cattccgtca
      661 gcattctggg tctgtttggt acttcttctc acgctaaggt ggcggtgtga tatgcacaat
      721 ggctaaaaag cagggagagc tggaaagaaa caaggacaga gacagaggcc aagtcaacca
      781 gaccaattcc cagaggaagc aaagaaacca ttacagagac tacaaggggg aagggaagga
      841 gagatgaatt agcttcccct gtaaacctta gaacccagct gttgccaggg caacggggca
      901 atacctgtct cttcagagga gatgaagttg ccagggtaac tacatcctgt ctttctcaag
      961 gaccatccca gaatgtggca cccactagcc gttaccatag caactgcctc tttgccccac
     1021 ttaatcccat cccgtctgtt aaaagggccc tatagttgga ggtgggggag gtaggaagag
     1081 cgatgatcac ttgtggacta agtttgttcg catccccttc tccaaccccc tcagtacatc
     1141 accctggggg aacagggtcc acttgctcct gggcccacac agtcctgcag tattgtgtat
     1201 ataaggccag ggcaaagagg agcaggtttt aaagtgaaag gcaggcaggt gttggggagg
     1261 cagttaccgg ggcaacggga acagggcgtt tcggaggtgg ttgccatggg gacctggatg
     1321 ctgacgaagg ctcgcgaggc tgtgagcagc cacagtgccc tgctcagaag ccccaagctc
     1381 gtcagtcaag ccggttctcc gtttgcactc aggagcacgg gcaggcgagt ggcccctagt
     1441 tctgggggca gcgggggatc cgccaccatg tcgcggaggc catggcttct tgccctagcg
     1501 ctggcagtgg cgctggcggc cggcagcgca ggagccgccc acatcgtgat ggtggacgcc
     1561 tacaagccca ccaagtcgac tggcagtgac gcgacggtgc cggtcgcgac tcaggatggc
     1621 cccgactacg ttttccaccg tgcccacgag cgcatgctct tccaaacctc atacactctt
     1681 gagaacaatg gttctgttat ttgcatcccg aacaacggcc agtgcttctg cttggcttgg
     1741 cttaaatcca acggaacaaa tgccgagaag ttggctgcca acattctgca gtggattact
     1801 tttgcgcttt cagcgctctg cctgatgttc tacggctacc agacctggaa gtctacttgc
     1861 ggctgggagg agatttacgt ggccacgatc gagatgatca agttcatcat cgagtatttc
     1921 catgagtttg acagtcctgc gatgctgttc ctgtacggag gcaacattac cccatggctt
     1981 cgttacgccg agtggctgct gacatgccct gtcattctta tccatctgag caacatcacc
     2041 ggtctgagtg aggcatataa taagcgtacc atggctctgc tggtgtcaga cctgggcact
     2101 atctgcatgg gcgtgacagc cgctctggcc actgggtggg tgaagtggct gttttactgt
     2161 atcggcctgg tgtacggcac ccagacattc tacaacgcag gaatcatcta cgtggaggcg
     2221 taccacaccg tgccgaaggg ccggtgtcgc caggtggtga ctggcatggc ttggctcttc
     2281 ttcgtatcat ggggtatgtt ccccatcctg ttcatcctcg gccccgaggg cttcggcgtc
     2341 ctgagcgtgg ctgggtccac tattggccac accatcgccg acctgctgtc caagaatatt
     2401 tggggactgc tcgggcactt cctgcgcatc aaaatccacg agcatatcct catccacggc
     2461 gacattcgca agaccaccaa attgaacatt ggtggcactg agattgaggt cgagacgctg
     2521 gtggaggacg aggccgaggc tggcgcggtc gcggccgcca agagcaggat caccagcgag
     2581 ggcgagtaca tccccctgga ccagatcgac atcaacgtgg tgagcaaggg cgaggagctg
     2641 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2701 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     2761 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc
     2821 ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     2881 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2941 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     3001 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     3061 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     3121 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     3181 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg
     3241 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     3301 gggatcactc tcggcatgga cgagctgtac aagtaataag aattcgatat caagcttatc
     3361 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3421 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3481 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     3541 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     3601 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     3661 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3721 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     3781 ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     3841 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     3901 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     3961 cgagcgctgc tcgagagatc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     4021 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     4081 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     4141 ggcaggacag caagggggag gattgggaag acaatagcag gcatgcacgt gcggaccgag
     4201 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     4261 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     4321 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     4381 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     4441 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4501 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4561 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4621 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4681 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4741 gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     4801 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4861 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4921 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     4981 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     5041 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     5101 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     5161 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5221 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5281 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5341 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5401 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5461 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5521 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5581 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5641 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5701 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5761 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5821 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5881 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5941 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6001 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6061 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6121 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6181 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6241 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6301 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6361 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6421 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     6481 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6541 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6601 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6661 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6721 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6781 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6841 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6901 cctttttacg gttcctggcc ttttgctggc cttttgctca catgt 
//
