LOCUS       sequence-219850-        6357 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  AAV expression of Cre recombinase-activated jRCaMP1a, a red 
            fluorescent calcium sensor protein, from CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6357)
  AUTHORS   Dana H, Mohar B, Sun Y, Narayan S, Gordus A, Hasseman JP, Tsegaye G,
            Holt GT, Hu A, Walpita D, Patel R, Macklin JJ, Bargmann CI, Ahrens 
            MB, Schreiter ER, Jayaraman V, Looger LL, Svoboda K, Kim DS
  TITLE     Sensitive red protein calcium indicators for imaging neural 
            activity.
  JOURNAL   eLife. 2016 Mar 24;5. pii: e12727. doi: 10.7554/eLife.12727.
  PUBMED    27011354
REFERENCE   2  (bases 1 to 6357)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "eLife."; 
            date: "2016-03-24"; volume: "5. pii: e12727. doi"; pages: " 
            10.7554/eLife.12727"
COMMENT     Sequence Label: pAAV.CAG.Flex.NES-jRCaMP1a.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..6357
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     repeat_region   422..551
                     /label=AAV2 ITR
     rep_origin      623..1078
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(710..729)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     920..941
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1104..1208
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1209..1277,1278..2069)
                     /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(1427..1446)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2240..2828
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2729..2748
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3065..3194
                     /label=AAV2 ITR
     polyA_signal    3356..3477
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3393..3412)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3447..3466
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    3520..4108
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3626..3637
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4035..4055
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    4150..4183
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4254..4287
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             4370..4387
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4391..4423
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(4427..4435,4436..4450)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             4478..4534
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     protein_bind    complement(5700..5733)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(5796..5829)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(5864..5883)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(5945..5967)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 cccccgcccc catcgctgca caaaataatt aaaaaataaa taaatacaaa attgggggtg
       61 gggagggggg ggagatgggg agagtgaagc agaacgtggg gctcacctcg accatggtaa
      121 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact
      181 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca
      241 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
      301 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
      361 gtactcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac
      421 aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
      481 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
      541 cgagcgcgca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
      601 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
      661 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
      721 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
      781 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
      841 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
      901 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
      961 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     1021 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     1081 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1141 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     1201 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     1261 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     1321 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     1381 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     1441 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     1501 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     1561 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     1621 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     1681 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     1741 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     1801 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     1861 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     1921 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     1981 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     2041 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     2101 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     2161 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     2221 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2281 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2341 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2401 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2461 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2521 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2581 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2641 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     2701 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     2761 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     2821 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     2881 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     2941 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3001 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     3061 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac
     3121 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
     3181 cactaggggt tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg
     3241 ctctaggaag atccttatcg attttaccac atttgtagag gttttacttg ctttaaaaaa
     3301 cctcccacat ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3361 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3421 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     3481 tgtctgctcg aagcggccgc ccgggtcgac ggtatcgatg cggggaggcg gcccaaaggg
     3541 agatccgact cgtctgaggg cgaaggcgaa gacgcggaag aggccgcaga gccggcagca
     3601 ggccgcggga aggaaggtcc gctggattga gggccgaagg gacgtagcag aaggacgtcc
     3661 cgcgcagaat ccaggtggca acacaggcga gcagccaagg aaaggacgat gatttccccg
     3721 acaacaccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc
     3781 aggcggcgat gagttccgcc gtggcaatag ggagggggaa agcgaaagtc ccggaaagga
     3841 gctgacaggt ggtggcaatg ccccaaccag tgggggttgc gtcagcaaac acagtgcaca
     3901 ccacgccacg ttgcctgaca acgggccaca actcctcata aagagacagc aaccaggatt
     3961 tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac aaaggcatta
     4021 aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca gtcaatcttt
     4081 cacaaatttt gtaatccaga ggttgattat cgataagctt gatatcgaat tcgaacctct
     4141 tcgagggaca taacttcgta taatgtatgc tatacgaagt tatgggtcga tggtgagatc
     4201 tggactagag ggtcgatggt gatgcttgga tcaaaatagg aagaatccca atgataactt
     4261 cgtataaagt atcctatacg aagttatact aggatccgcc accatgctgc agaacgagct
     4321 tgctcttaag ttggctggac ttgatattaa caagactgga ggaggttctc atcatcatca
     4381 tcatcatggt atggctagca tgactggtgg acagcaaatg ggtcgggatc tgtacgacga
     4441 tgacgataag gatctcgcaa caatggtcga ctcatcacgt cgtaagtgga ataagacagg
     4501 tcacgcagtc agagctatag gtcggctgag ctcagcgatc aactgcgaaa tgatgtaccc
     4561 agcggatggt ggtctgcgtg gttacactca catggcgctg aaagttgatg gcggcggtca
     4621 cctgtcctgt tctttcgtga ccacctaccg ctccaaaaag actgtcggca acattaagat
     4681 gcctgccatt catagcgtca gccaccgtct ggagcgcctg gaggagagcg ataacgaaat
     4741 gtttgtcgta cagcgtgaac acgcagttgc caagtttgtg ggcctgggtg gtggcggcgg
     4801 taccggaggg agcatgaact ccctgatcaa ggagaacatg cgtatgaaag tggttctgga
     4861 aggctccgta aacggccacc agttcaaatg cactggtgaa ggcgaaggca acccgtatat
     4921 gggcacccag actatgcgta tcaaagtgat cgagggtggt ccgctgccgt ttgcgttcga
     4981 catcctggcg acgtccttta tgtatggctc ccgtaccttc atcaaatatc cgaaaggcat
     5041 cccggatttc tttaagcagt ccttcccgga aggttttacc tgggaacgtg tgacccgtta
     5101 cgaagacggc ggcgtaatta ccgttatgca agacacgtct ctggaggatg gctgcctggt
     5161 gtatcacgtg caggttcgcg gtgtgaactt cccgagcaat ggtgctgtaa tgcaaaagaa
     5221 aaccaaaggt tgggagccta cggactccca aaccactgaa gagcagatcg cagaatttaa
     5281 agaggctttc tccctatttg acaaggacgg ggatgggaca ataacaacca aggagctggg
     5341 gacggtgatg cggtctctgg ggcagaaccc cacagaagca gagctgcagg acatgatcaa
     5401 tgaagtagat gccgacggtg acggcacaat cgacttccct gagttcctga ttatgatggc
     5461 aagaaaaatg aaatacacag acagtgaaga agaaattaga gaagcgttcg gcgtgtttga
     5521 taaggatggc aatggctaca tcagtgcagc agagcttcgc cacgtgatga caaaccttgg
     5581 agagaagtta acagatgaag aggttgatga aatgatcagg gaagcagaca gcgatgggga
     5641 tggtcaggta aactacgaag agtttgtaca aatgatgaca gcgaagtgaa agctctagta
     5701 taacttcgta tagcatacat tatacgaagt tatgggtcga tggtgatgct tggcaattcg
     5761 ggtcgatggt gaagcattgg tcttcctatt ttgatataac ttcgtatagg atactttata
     5821 cgaagttatg gggatccaat tctttgccaa aatgatgaga cagcacaata accagcacgt
     5881 tgcccaggag ctgtaggaaa aagaagaagg catgaacatg gttagcagag gctctagagc
     5941 cgccggtcac acgccagaag ccgaaccccg ccctgccccg tcccccccga aggcagccgt
     6001 ccccccgcgg acagccccga gccgctcccc ccgcacaaag ggccctcccg gagcccctca
     6061 aggctttcac gcagccacag aaaagaaaca agccgtcatt aaaccaagcg ctaattacag
     6121 cccggaggag aagggccgtc ccgcccgctc acctgtggga gtaacgcggt cagtcagagc
     6181 cggggcgggc ggcgcgaggc ggcggcggag cggggcacgg ggcgaaggca gcgcgcagcg
     6241 actcccgccc gccgcgcgct tcgcttttta tagggccgcc gccgccgccg cctcgccata
     6301 aaaggaaact ttcggagcgc gccgctctga ttggctgccg ccgcacctct ccgcctc
//
