LOCUS       sequence-214502-        6155 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of Cre recombinase-activated 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 6155)
  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 6155)
  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-jRGECO1b.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..6155
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   71..200
                     /label=AAV2 ITR
     promoter        282..729
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    759..792
                     /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    855..888
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(1366..1385)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(2081..2137)
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     CDS             complement(join(2165..2179,2180..2188))
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             complement(2192..2224)
                     /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(2228..2245)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(2328..2361)
                     /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(2432..2465)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2507..3095
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2560..2580)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2978..2989)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3138..3259
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3149..3168)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3203..3222
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3421..3550
                     /label=AAV2 ITR
     rep_origin      complement(3949..4537)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4029..4048)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4708..5499,5500..5568))
                     /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     5331..5350
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5569..5673)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5699..6154)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5836..5857)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6048..6067
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg
      121 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
      181 ctccatcact aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta
      241 gccatgctct aggaagatct ctgcagaggg ccctgcgtat gagtgcaagt gggttttagg
      301 accaggatga ggcggggtgg gggtgcctac ctgacgaccg accccgaccc actggacaag
      361 cacccaaccc ccattcccca aattgcgcat cccctatcag agagggggag gggaaacagg
      421 atgcggcgag gcgcgtgcgc actgccagct tcagcaccgc ggacagtgcc ttcgcccccg
      481 cctggcggcg cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc actcgccggt
      541 cccccgcaaa ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc
      601 ggaccgcacc acgcgaggcg cgagataggg gggcacgggc gcgaccatct gcgctgcggc
      661 gccggcgact cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt gtcgtgcctg
      721 agagcgcagt cgaattcaag ctgctagcaa ggatccccat aacttcgtat aaagtatcct
      781 atacgaagtt atatcaaaat aggaagacca atgcttcacc atcgacccga attgccaagc
      841 atcaccatcg acccataact tcgtataatg tatgctatac gaagttatac tagagctttc
      901 acttcgctgt catcatttgt acaaactctt cgtagtttac ctgaccatcc ccatcgatgt
      961 ctgctaccct gatcatttca tcaacctcct catctgttaa cttctctcca aggtctgtca
     1021 tcacgtggcg aagctctgct gcgccgatgt agccattgcc gtccttatca aacacgcgga
     1081 acgcttcgcg aatttcctct tcactgtctg tgtcattcat ttttcttgcc atcatcgtca
     1141 ggaactcagg gaagtcgaat gtgccgtcac cgtcggcatc tacttcattg atcatgtcct
     1201 gcagctctgc ttctgtgggg ttctgcccca gcgatcgcat caccgtccca agctccttgg
     1261 ttgttatcgt cccatccccg tccttgtcaa agcaggagaa agcctcttta aattctgcga
     1321 tctgctcttc agtcagttgg tcacgcgtag cctcccagcc catggtcttc ttctgcatta
     1381 cggggccgtc cggagggaag ttggtgccgc gcagcttcac cttgtagatg aatacgccgt
     1441 cctgcaggga ggagtcctgg ttcacgtgaa taatgccgcc gtcctcgaag ttcatcacgc
     1501 gctcccacct gaagccctcg gggaaggaca gcttgaagta gtcggggatg tcggctgggt
     1561 gcttaatgta ggccttggag ccgtacatga actgagggga caggatgtcc caggcgaagg
     1621 gcagggggcc acccttggtc accttcagct tagcggtctg aaaggcctcg taggggcggc
     1681 cctcgccctc gccctcgatc tcgaactcgt ggccgttcac ggagccctcc atgtgcacct
     1741 tgaagcgcat gaactccttg atgatggcca tgttatcctc ctcgcccttg ctcaccagac
     1801 tcccgcctgt acctcccttg tacagctcgt ccatgccgcc ggtggagtgg cggccctcgg
     1861 cacattcgca ctgttccacg atggtgtagt cctcgttgtg ggagacgatg tccaacttga
     1921 tgtccacctt gtaggcgccg ggcagctgca cgggcttctt ggccttgtag gtagtcttga
     1981 cctcggcggc gtagtggccg ccgtccttca gcctcagccc cttcttgatc tcgctcttaa
     2041 gggcgccgtc ctcggggtac atccgctcgg aaaccacggg tgagctcagc cgacctatag
     2101 ctctgactgc gtgacctgcc ttattccact tacgacgtga tgagtcgacc attgttgcga
     2161 gatccttatc gtcatcgtcg tacagatccc gacccatttg ctgtccacca gtcatgctag
     2221 ccataccatg atgatgatga tgatgagaac ctcctccagt cttgttaata tcaagtccag
     2281 ccaacttaag agcaagctcg ttctgcagca tggtggcgga tcctagtata acttcgtata
     2341 ggatacttta tacgaagtta tcattgggat tcttcctatt ttgatccaag catcaccatc
     2401 gaccctctag tccagatctc accatcgacc cataacttcg tatagcatac attatacgaa
     2461 gttatgtccc tcgaagaggt tcgaattcga tatcaagctt atcgataatc aacctctgga
     2521 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2581 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2641 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2701 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     2761 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     2821 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     2881 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     2941 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3001 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3061 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac ccgggcggcc
     3121 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3181 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3241 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3301 ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa
     3361 ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca
     3421 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     3481 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     3541 gagcgcgcag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     3601 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3661 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgcc cccttcgctt
     3721 tcttcccytc ctttctgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3781 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3841 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3901 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3961 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4021 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4081 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4141 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4201 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4261 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4321 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4381 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4441 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4501 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4561 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4621 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4681 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4741 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4801 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4861 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4921 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4981 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5041 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5101 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5161 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5221 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5281 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5341 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5401 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5461 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5521 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5581 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5641 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     5701 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     5761 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     5821 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     5881 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     5941 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     6001 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     6061 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     6121 cccgccgcgc ttaatgcgcc gctacagggc gcgtc
//
