LOCUS       sequence-189704-        6412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of Cre recombinase-activated jRCaMP1a, a red 
            fluorescent calcium sensor protein, from CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6412)
  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 6412)
  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: "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..6412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        441..718
                     /label=chicken beta-actin promoter
     primer_bind     997..1019
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1081..1100
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1135..1168
                     /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)."
     protein_bind    1231..1264
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2430..2486)
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     CDS             complement(join(2514..2528,2529..2537))
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             complement(2541..2573)
                     /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             complement(2577..2594)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(2677..2710)
                     /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)."
     protein_bind    complement(2781..2814)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2856..3444
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2909..2929)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3327..3338)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3487..3608
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3498..3517)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3552..3571
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3770..3899
                     /label=AAV2 ITR
     rep_origin      complement(4136..4724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4216..4235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4895..5686,5687..5755))
                     /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     5518..5537
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5756..5860)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5886..6341)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6023..6044)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6235..6254
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagagta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      421 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
      481 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
      541 gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
      601 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      661 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      721 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
      781 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      841 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
      901 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggc tgtccgcggg
      961 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1021 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1081 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattggat ccccataact
     1141 tcgtataaag tatcctatac gaagttatat caaaatagga agaccaatgc ttcaccatcg
     1201 acccgaattg ccaagcatca ccatcgaccc ataacttcgt ataatgtatg ctatacgaag
     1261 ttatactaga gctttcactt cgctgtcatc atttgtacaa actcttcgta gtttacctga
     1321 ccatccccat cgctgtctgc ttccctgatc atttcatcaa cctcttcatc tgttaacttc
     1381 tctccaaggt ttgtcatcac gtggcgaagc tctgctgcac tgatgtagcc attgccatcc
     1441 ttatcaaaca cgccgaacgc ttctctaatt tcttcttcac tgtctgtgta tttcattttt
     1501 cttgccatca taatcaggaa ctcagggaag tcgattgtgc cgtcaccgtc ggcatctact
     1561 tcattgatca tgtcctgcag ctctgcttct gtggggttct gccccagaga ccgcatcacc
     1621 gtccccagct ccttggttgt tattgtccca tccccgtcct tgtcaaatag ggagaaagcc
     1681 tctttaaatt ctgcgatctg ctcttcagtg gtttgggagt ccgtaggctc ccaacctttg
     1741 gttttctttt gcattacagc accattgctc gggaagttca caccgcgaac ctgcacgtga
     1801 tacaccaggc agccatcctc cagagacgtg tcttgcataa cggtaattac gccgccgtct
     1861 tcgtaacggg tcacacgttc ccaggtaaaa ccttccggga aggactgctt aaagaaatcc
     1921 gggatgcctt tcggatattt gatgaaggta cgggagccat acataaagga cgtcgccagg
     1981 atgtcgaacg caaacggcag cggaccaccc tcgatcactt tgatacgcat agtctgggtg
     2041 cccatatacg ggttgccttc gccttcacca gtgcatttga actggtggcc gtttacggag
     2101 ccttccagaa ccactttcat acgcatgttc tccttgatca gggagttcat gctccctccg
     2161 gtaccgccgc caccacccag gcccacaaac ttggcaactg cgtgttcacg ctgtacgaca
     2221 aacatttcgt tatcgctctc ctccaggcgc tccagacggt ggctgacgct atgaatggca
     2281 ggcatcttaa tgttgccgac agtctttttg gagcggtagg tggtcacgaa agaacaggac
     2341 aggtgaccgc cgccatcaac tttcagcgcc atgtgagtgt aaccacgcag accaccatcc
     2401 gctgggtaca tcatttcgca gttgatcgct gagctcagcc gacctatagc tctgactgcg
     2461 tgacctgtct tattccactt acgacgtgat gagtcgacca ttgttgcgag atccttatcg
     2521 tcatcgtcgt acagatcccg acccatttgc tgtccaccag tcatgctagc cataccatga
     2581 tgatgatgat gatgagaacc tcctccagtc ttgttaatat caagtccagc caacttaaga
     2641 gcaagctcgt tctgcagcat ggtggcggat cctagtataa cttcgtatag gatactttat
     2701 acgaagttat cattgggatt cttcctattt tgatccaagc atcaccatcg accctctagt
     2761 ccagatctca ccatcgaccc ataacttcgt atagcataca ttatacgaag ttatgtccct
     2821 cgaagaggtt cgaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
     2881 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     2941 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3001 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3061 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3121 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3181 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3241 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     3301 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3361 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3421 tctccctttg ggccgcctcc ccgcatcgat accgtcgacc cgggcggccg cttcgagcag
     3481 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     3541 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     3601 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     3661 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta
     3721 gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta
     3781 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     3841 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     3901 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3961 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4021 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4081 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4141 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4201 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4261 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4321 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4381 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4441 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4501 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4561 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4621 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4681 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4741 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4801 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4861 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4921 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4981 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5041 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5101 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5161 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5221 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5281 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5341 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5401 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5461 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5521 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5581 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5641 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5701 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5761 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5821 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5881 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     5941 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     6001 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     6061 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     6121 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     6181 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     6241 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     6301 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
     6361 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc ag
//
