LOCUS       sequence-197165-        5684 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of jRGECO1b, 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 5684)
  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 5684)
  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-jRGECO1b.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..5684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   4..133
                     /label=AAV2 ITR
     promoter        215..662
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             766..783
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             787..819
                     /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(823..831,832..846)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             874..930
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     1626..1645
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2125..2713
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2178..2198)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2596..2607)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2716..2732)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2717..2733)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2762..2883
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2773..2792)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2827..2846
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3045..3174
                     /label=AAV2 ITR
     rep_origin      complement(3411..3999)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3491..3510)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4170..4961,4962..5030))
                     /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     4793..4812
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5031..5135)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5161..5616)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5298..5319)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5510..5529
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctaggaaga tctctgcaga gggccctgcg tatgagtgca agtgggtttt aggaccagga
      241 tgaggcgggg tgggggtgcc tacctgacga ccgaccccga cccactggac aagcacccaa
      301 cccccattcc ccaaattgcg catcccctat cagagagggg gaggggaaac aggatgcggc
      361 gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt gccttcgccc ccgcctggcg
      421 gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc ggtcccccgc
      481 aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca gccggaccgc
      541 accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc ggcgccggcg
      601 actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc ctgagagcgc
      661 agtcgaattc aagctgctag caaggatcca cccgccacca tgctgcagaa cgagcttgct
      721 cttaagttgg ctggacttga tattaacaag actggaggag gttctcatca tcatcatcat
      781 catggtatgg ctagcatgac tggtggacag caaatgggtc gggatctgta cgacgatgac
      841 gataaggatc tcgcaacaat ggtcgactca tcacgtcgta agtggaataa ggcaggtcac
      901 gcagtcagag ctataggtcg gctgagctca cccgtggttt ccgagcggat gtaccccgag
      961 gacggcgccc ttaagagcga gatcaagaag gggctgaggc tgaaggacgg cggccactac
     1021 gccgccgagg tcaagactac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     1081 aaggtggaca tcaagttgga catcgtctcc cacaacgagg actacaccat cgtggaacag
     1141 tgcgaatgtg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggaggt
     1201 acaggcggga gtctggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     1261 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     1321 gagggcgagg gccgccccta cgaggccttt cagaccgcta agctgaaggt gaccaagggt
     1381 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     1441 tacattaagc acccagccga catccccgac tacttcaagc tgtccttccc cgagggcttc
     1501 aggtgggagc gcgtgatgaa cttcgaggac ggcggcatta ttcacgtgaa ccaggactcc
     1561 tccctgcagg acggcgtatt catctacaag gtgaagctgc gcggcaccaa cttccctccg
     1621 gacggccccg taatgcagaa gaagaccatg ggctgggagg ctacgcgtga ccaactgact
     1681 gaagagcaga tcgcagaatt taaagaggct ttctcctgct ttgacaagga cggggatggg
     1741 acgataacaa ccaaggagct tgggacggtg atgcgatcgc tggggcagaa ccccacagaa
     1801 gcagagctgc aggacatgat caatgaagta gatgccgacg gtgacggcac attcgacttc
     1861 cctgagttcc tgacgatgat ggcaagaaaa atgaatgaca cagacagtga agaggaaatt
     1921 cgcgaagcgt tccgcgtgtt tgataaggac ggcaatggct acatcggcgc agcagagctt
     1981 cgccacgtga tgacagacct tggagagaag ttaacagatg aggaggttga tgaaatgatc
     2041 agggtagcag acatcgatgg ggatggtcag gtaaactacg aagagtttgt acaaatgatg
     2101 acagcgaagt gaaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     2161 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     2221 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     2281 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2341 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2401 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2461 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     2521 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     2581 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     2641 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     2701 gccgcctccc cgcatcgata ccgtcgacct cgacccgggc ggccgcttcg agcagacatg
     2761 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2821 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2881 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     2941 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct tcctagagca
     3001 tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat
     3061 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     3121 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcat
     3181 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3241 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3301 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3361 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3421 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3481 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3541 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3601 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3661 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3721 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3781 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3841 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3901 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3961 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4021 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4081 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4141 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4201 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4261 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4321 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4381 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4441 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4501 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4561 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4621 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4681 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4741 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4801 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4861 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4921 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4981 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5041 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5101 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5161 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     5221 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     5281 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     5341 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     5401 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     5461 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     5521 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     5581 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
     5641 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgc
//
