LOCUS       sequence-487629-        5950 bp    DNA     circular SYN 01-OCT-2025
DEFINITION  AAV expression of Cre recombinase-activated jRCaMP1b, a red 
            fluorescent calcium sensor protein, from the Synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5950)
  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 5950)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 1, 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.Syn.Flex.NES-jRCaMP1b.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..5950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    27..148
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(64..83)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     118..137
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     177..193
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     178..194
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    197..785
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             303..314
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     712..732
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    827..860
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    931..964
                     /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             1047..1064
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1068..1100
                     /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(1104..1112,1113..1127)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     protein_bind    complement(2377..2410)
                     /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(2473..2506)
                     /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)."
     promoter        complement(2536..2983)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   3065..3194
                     /label=AAV2 ITR
     rep_origin      3266..3721
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3353..3372)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3563..3584
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3747..3851
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3852..3920,3921..4712)
                     /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(4070..4089)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4883..5471
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5372..5391
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
       61 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
      121 gtttgtccaa actcatcaat gtatcttatc atgtctgctc gaagcggccg cccgggtcga
      181 ggtcgacggt atcgatgcgg ggaggcggcc caaagggaga tccgactcgt ctgagggcga
      241 aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg aaggtccgct
      301 ggattgaggg ccgaagggac gtagcagaag gacgtcccgc gcagaatcca ggtggcaaca
      361 caggcgagca gccaaggaaa ggacgatgat ttccccgaca acaccacgga attgtcagtg
      421 cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg
      481 gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt ggcaatgccc
      541 caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg cctgacaacg
      601 ggccacaact cctcataaag agacagcaac caggatttat acaaggagga gaaaatgaaa
      661 gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc acatagcgta
      721 aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta atccagaggt
      781 tgattatcga taagcttgat atcgaattcg aacctcttcg agggacataa cttcgtataa
      841 tgtatgctat acgaagttat gggtcgatgg tgagatctgg actagagggt cgatggtgat
      901 gcttggatca aaataggaag aatcccaatg ataacttcgt ataaagtatc ctatacgaag
      961 ttatactagg atccgccacc atgctgcaga acgagcttgc tcttaagttg gctggacttg
     1021 atattaacaa gactggagga ggttctcatc atcatcatca tcatggtatg gctagcatga
     1081 ctggtggaca gcaaatgggt cgggatctgt acgacgatga cgataaggat ctcgcaacaa
     1141 tggtcgactc atcgcgacgt aagtggaata agtggggtca cgcagtcaga gctataggtc
     1201 ggctgagctc agcgaacaac accgaaatga tgtacccagc ggatggtggt ctgcgtggtt
     1261 acactcacat ggcgctgaaa gttgatggcg gcggtcacct gtcctgttct ttcgtgacca
     1321 cctaccgctc caaaaagact gtcggcaaca ttaagatgcc tgccattcat tacgtcagcc
     1381 accgtctgga gcgcctggag gagagcgata acgaaatgtt tgtcgtacag cgtgaacacg
     1441 cagttgccaa gtttgtgggc ctgggtggtg gcggcggtac cggagggagc atgaactccc
     1501 tgatcaagga gaacatgcgt atgaaagtgg ttctggaagg ctccgtaaac ggccaccagt
     1561 tcaaatgcac tggtgaaggc gaaggcaacc cgtatatggg cacccagact atgcgtatca
     1621 aagtgatcga gggtggtccg ctgccgtttg cgttcgacat cctggcgacg tcctttatgt
     1681 atggctcccg taccttcatc aaatatccga aaggcatccc ggatttcttt aagcagtcct
     1741 tcccggaagg ttttacctgg gaacgtgtga cccgttacga agacggcggc gtaattaccg
     1801 ttatgcaaga cacgtctctg gaggatggct gcctggtgta tcacgtgcag gttcgcggtg
     1861 tgaacttccc gagcaatggt gctgtaatgc aaaagaaaac caaaggttgg gagcctacgg
     1921 actcccaact gactgaagag cagatcgcag aatttaaaga ggctttctcc ctatttgaca
     1981 aggacgggga tgggacaata acaaccaagg agatggggac ggtgatgcgg tctctggggc
     2041 agaaccccac agaagcagag ctgcaggaca tgatcaatga agtagatgcc gacggtgacg
     2101 gcacaatcga cttccctgag ttcctgatta tgatggcagg caaaatgaaa tacacagaca
     2161 gtgaagaaga aattagagaa gcgttcggcg tgtttgataa ggatggcaat ggctacatca
     2221 gtgcagcaga gcttcgccac gtgatgacaa accttggaga gaagttaaca gatgaagagg
     2281 ttgatgaaat gatcagggaa gcagacagcg atggggatgg tcaggtaaac tacgaagagt
     2341 ttgtacaaat gatgacagcg aagtgaaagc tctagtataa cttcgtatag catacattat
     2401 acgaagttat gggtcgatgg tgatgcttgg caattcgggt cgatggtgaa gcattggtct
     2461 tcctattttg atataacttc gtataggata ctttatacga agttatgggg atccttgcta
     2521 gcagcttgaa ttcgactgcg ctctcaggca cgacacgact cctccgctgc ccaccgcaga
     2581 ctgaggcagc gctgagtcgc cggcgccgca gcgcagatgg tcgcgcccgt gcccccctat
     2641 ctcgcgcctc gcgtggtgcg gtccggctgg gccggcggcg gcgcggacgc gaccaaggtg
     2701 gccgggaagg ggagtttgcg ggggaccggc gagtgacgtc agcgcgcctt cagtgctgag
     2761 gcggcggtgg cgcgcgccgc caggcggggg cgaaggcact gtccgcggtg ctgaagctgg
     2821 cagtgcgcac gcgcctcgcc gcatcctgtt tcccctcccc ctctctgata ggggatgcgc
     2881 aatttgggga atgggggttg ggtgcttgtc cagtgggtcg gggtcggtcg tcaggtaggc
     2941 acccccaccc cgcctcatcc tggtcctaaa acccacttgc actcatacgc agggccctct
     3001 gcagagatct tcctagagca tggctacgta gataagtagc atggcgggtt aatcattaac
     3061 tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     3121 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     3181 gagcgagcgc gcagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     3241 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3301 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3361 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3421 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3481 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3541 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3601 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3661 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     3721 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3781 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3841 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3901 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3961 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4021 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4081 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4141 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4201 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4261 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4321 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4381 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4441 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4501 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4561 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4621 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4681 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4741 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4801 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4861 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4921 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4981 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5041 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5101 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5161 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5221 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5281 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5341 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5401 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5461 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5521 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5581 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5641 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5701 aatgcagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcggcctca
     5761 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag
     5821 ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa gatccttatc
     5881 gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca tctccccctg
     5941 aacctgaaac
//
