LOCUS       sequence-455105-        5977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of Cre recombinase-activated jRGECO1a, 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 5977)
  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 5977)
  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.Flex.NES-jRGECO1a.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..5977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    269..390
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(306..325)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     360..379
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     419..435
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     420..436
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    439..1027
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             545..556
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     954..974
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1069..1102
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1173..1206
                     /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             1289..1306
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1310..1342
                     /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(1346..1354,1355..1369)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1397..1453
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     2149..2168
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2646..2679)
                     /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(2742..2775)
                     /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(2805..3252)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   3334..3463
                     /label=AAV2 ITR
     rep_origin      3535..3990
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3622..3641)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3832..3853
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4016..4120
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4121..4189,4190..4981)
                     /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(4339..4358)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5152..5740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5641..5660
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcggcct cagtgagcga
       61 gcgagcgcgc agagagggag tggccaactc catcactagg ggttccttgt agttaatgat
      121 taacccgcca tgctacttat ctacgtagcc atgctctagg aagatcctta tcgattttac
      181 cacatttgta gaggttttac ttgctttaaa aaacctccca catctccccc tgaacctgaa
      241 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
      301 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
      361 tggtttgtcc aaactcatca atgtatctta tcatgtctgc tcgaagcggc cgcccgggtc
      421 gaggtcgacg gtatcgatgc ggggaggcgg cccaaaggga gatccgactc gtctgagggc
      481 gaaggcgaag acgcggaaga ggccgcagag ccggcagcag gccgcgggaa ggaaggtccg
      541 ctggattgag ggccgaaggg acgtagcaga aggacgtccc gcgcagaatc caggtggcaa
      601 cacaggcgag cagccaagga aaggacgatg atttccccga caacaccacg gaattgtcag
      661 tgcccaacag ccgagcccct gtccagcagc gggcaaggca ggcggcgatg agttccgccg
      721 tggcaatagg gagggggaaa gcgaaagtcc cggaaaggag ctgacaggtg gtggcaatgc
      781 cccaaccagt gggggttgcg tcagcaaaca cagtgcacac cacgccacgt tgcctgacaa
      841 cgggccacaa ctcctcataa agagacagca accaggattt atacaaggag gagaaaatga
      901 aagccatacg ggaagcaata gcatgataca aaggcattaa agcagcgtat ccacatagcg
      961 taaaaggagc aacatagtta agaataccag tcaatctttc acaaattttg taatccagag
     1021 gttgattatc gataagcttg atatcgaatt cgaacctctt cgagggacat aacttcgtat
     1081 aatgtatgct atacgaagtt atgggtcgat ggtgagatct ggactagagg gtcgatggtg
     1141 atgcttggat caaaatagga agaatcccaa tgataacttc gtataaagta tcctatacga
     1201 agttatacta ggatccgcca ccatgctgca gaacgagctt gctcttaagt tggctggact
     1261 tgatattaac aagactggag gaggttctca tcatcatcat catcatggta tggctagcat
     1321 gactggtgga cagcaaatgg gtcgggatct gtacgacgat gacgataagg atctcgcaac
     1381 aatggtcgac tcatcacgtc gtaagtggaa taaggcaggt cacgcagtca gagctatagg
     1441 tcggctgagc tcacccgtgg tttccgagcg gatgtacccc gaggacggcg ccctgaagag
     1501 cgagatcaag aaggggctga ggctgaagga cggcggccac tacgccgccg aggtcaagac
     1561 cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacatcgtgg acatcaagtt
     1621 ggacatcgtg tcccacaacg aggactacac catcgtggaa cagtgcgaac gcgccgaggg
     1681 ccgccactcc accggcggca tggacgagct gtacaaggga ggtacaggcg ggagtctggt
     1741 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     1801 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     1861 ctacgaggcc tttcagaccg ctaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     1921 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacatta agcacccagc
     1981 cgacatcccc gactacttca agctgtcctt ccccgagggc ttcaggtggg agcgcgtgat
     2041 gaacttcgag gacggcggca ttattcacgt gaaccaggac tcctccctgc aggacggcgt
     2101 attcatctac aaggtgaagc tgcgcggcac caacttcccc cccgacggcc ccgtaatgca
     2161 gaagaagacc atgggctggg aggctacgcg tgacgacctg actgaagagc agatcgcaga
     2221 atttaaagag gctttctccc tatttgacaa ggacggggat gggacgataa caaccaagga
     2281 gcttgggacg gtgttccgat cgctggggca gaaccccaca gaagcagagc tgcaggacat
     2341 gatcaatgaa gtagatgccg acggtgacgg cacattcgac ttccctgagt tcctgacgat
     2401 gatggcaaga aaaatgaatg acacagacag tgaagaggaa attcgcgaag cgttccgcgt
     2461 gtttgataag gacggcaatg gctacatcgg cgcagcagag cttcgccacg tgatgacaga
     2521 ccttggagag aagttaacag atgaggaggt tgatgaaatg atcagggtag cagacatcga
     2581 tggggatggt caggtaaact acgaagagtt tgtacaaatg atgacagcga agtgaaagct
     2641 ctagtataac ttcgtatagc atacattata cgaagttatg ggtcgatggt gatgcttggc
     2701 aattcgggtc gatggtgaag cattggtctt cctattttga tataacttcg tataggatac
     2761 tttatacgaa gttatgggga tccttgctag cagcttgaat tcgactgcgc tctcaggcac
     2821 gacacgactc ctccgctgcc caccgcagac tgaggcagcg ctgagtcgcc ggcgccgcag
     2881 cgcagatggt cgcgcccgtg cccccctatc tcgcgcctcg cgtggtgcgg tccggctggg
     2941 ccggcggcgg cgcggacgcg accaaggtgg ccgggaaggg gagtttgcgg gggaccggcg
     3001 agtgacgtca gcgcgccttc agtgctgagg cggcggtggc gcgcgccgcc aggcgggggc
     3061 gaaggcactg tccgcggtgc tgaagctggc agtgcgcacg cgcctcgccg catcctgttt
     3121 cccctccccc tctctgatag gggatgcgca atttggggaa tgggggttgg gtgcttgtcc
     3181 agtgggtcgg ggtcggtcgt caggtaggca cccccacccc gcctcatcct ggtcctaaaa
     3241 cccacttgca ctcatacgca gggccctctg cagagatctt cctagagcat ggctacgtag
     3301 ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca
     3361 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3421 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctggcgt aatagcgaag
     3481 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     3541 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3601 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3661 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3721 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3781 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3841 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3901 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3961 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     4021 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4081 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4141 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4201 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4261 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4321 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4381 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4441 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4501 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4561 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4621 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4681 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4741 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4801 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4861 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4921 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4981 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5041 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5101 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5161 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5221 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5281 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5341 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5401 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5461 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5521 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5581 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5641 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5701 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5761 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5821 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     5881 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     5941 gcctctcccc gcgcgttggc cgattcatta atgcagc
//
