LOCUS       sequence-198187-        5657 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of 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 5657)
  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 5657)
  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.Syn.NES-jRCaMP1b.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..5657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             341..358
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             362..394
                     /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(398..406,407..421)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     misc_feature    1673..2261
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1726..1746)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2144..2155)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2264..2280)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2265..2281)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2310..2431
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2321..2340)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2375..2394
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2593..2722
                     /label=AAV2 ITR
     rep_origin      complement(2959..3547)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3039..3058)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3718..4509,4510..4578))
                     /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     4341..4360
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4579..4683)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4709..5164)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4846..4867)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5058..5077
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5236..5365
                     /label=AAV2 ITR
     promoter        join(5447..5657,1..237)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact
       61 ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg accgcaccac
      121 gcgaggcgcg agataggggg gcacgggcgc gaccatctgc gctgcggcgc cggcgactca
      181 gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg
      241 aattcaagct gctagcaagg atccacccgc caccatgctg cagaacgagc ttgctcttaa
      301 gttggctgga cttgatatta acaagactgg aggaggttct catcatcatc atcatcatgg
      361 tatggctagc atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa
      421 ggatctcgca acaatggtcg actcatcgcg acgtaagtgg aataagtggg gtcacgcagt
      481 cagagctata ggtcggctga gctcagcgaa caacaccgaa atgatgtacc cagcggatgg
      541 tggtctgcgt ggttacactc acatggcgct gaaagttgat ggcggcggtc acctgtcctg
      601 ttctttcgtg accacctacc gctccaaaaa gactgtcggc aacattaaga tgcctgccat
      661 tcattacgtc agccaccgtc tggagcgcct ggaggagagc gataacgaaa tgtttgtcgt
      721 acagcgtgaa cacgcagttg ccaagtttgt gggcctgggt ggtggcggcg gtaccggagg
      781 gagcatgaac tccctgatca aggagaacat gcgtatgaaa gtggttctgg aaggctccgt
      841 aaacggccac cagttcaaat gcactggtga aggcgaaggc aacccgtata tgggcaccca
      901 gactatgcgt atcaaagtga tcgagggtgg tccgctgccg tttgcgttcg acatcctggc
      961 gacgtccttt atgtatggct cccgtacctt catcaaatat ccgaaaggca tcccggattt
     1021 ctttaagcag tccttcccgg aaggttttac ctgggaacgt gtgacccgtt acgaagacgg
     1081 cggcgtaatt accgttatgc aagacacgtc tctggaggat ggctgcctgg tgtatcacgt
     1141 gcaggttcgc ggtgtgaact tcccgagcaa tggtgctgta atgcaaaaga aaaccaaagg
     1201 ttgggagcct acggactccc aactgactga agagcagatc gcagaattta aagaggcttt
     1261 ctccctattt gacaaggacg gggatgggac aataacaacc aaggagatgg ggacggtgat
     1321 gcggtctctg gggcagaacc ccacagaagc agagctgcag gacatgatca atgaagtaga
     1381 tgccgacggt gacggcacaa tcgacttccc tgagttcctg attatgatgg caggcaaaat
     1441 gaaatacaca gacagtgaag aagaaattag agaagcgttc ggcgtgtttg ataaggatgg
     1501 caatggctac atcagtgcag cagagcttcg ccacgtgatg acaaaccttg gagagaagtt
     1561 aacagatgaa gaggttgatg aaatgatcag ggaagcagac agcgatgggg atggtcaggt
     1621 aaactacgaa gagtttgtac aaatgatgac agcgaagtga aagcttatcg ataatcaacc
     1681 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     1741 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     1801 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     1861 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     1921 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     1981 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2041 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     2101 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     2161 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     2221 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg
     2281 acccgggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     2341 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     2401 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     2461 tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     2521 taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa
     2581 tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     2641 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     2701 cagtgagcga gcgagcgcgc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2761 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2821 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2881 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2941 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3001 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3061 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3121 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3181 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3241 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3301 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3361 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3421 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3481 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3541 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3601 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3661 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3721 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3781 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3841 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3901 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3961 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4021 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4081 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4141 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4201 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4261 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4321 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4381 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4441 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4501 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4561 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4621 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4681 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     4741 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     4801 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     4861 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     4921 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     4981 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     5041 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     5101 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     5161 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     5221 tcgctattac gccagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg
     5281 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
     5341 gccaactcca tcactagggg ttccttgtag ttaatgatta acccgccatg ctacttatct
     5401 acgtagccat gctctaggaa gatctctgca gagggccctg cgtatgagtg caagtgggtt
     5461 ttaggaccag gatgaggcgg ggtgggggtg cctacctgac gaccgacccc gacccactgg
     5521 acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg gggaggggaa
     5581 acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca gtgccttcgc
     5641 ccccgcctgg cggcgcg
//
