LOCUS       sequence-198193-        5684 bp    DNA     circular SYN 22-APR-2025
DEFINITION  AAV expression of 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 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 Apr 22, 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-jRGECO1a.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..5684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             354..371
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             375..407
                     /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(411..419,420..434)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             462..518
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     1214..1233
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1713..2301
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1766..1786)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2184..2195)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2304..2320)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2305..2321)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2350..2471
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2361..2380)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2415..2434
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2633..2762
                     /label=AAV2 ITR
     rep_origin      complement(2999..3587)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3079..3098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3758..4549,4550..4618))
                     /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     4381..4400
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4619..4723)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4749..5204)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4886..4907)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5098..5117
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5276..5405
                     /label=AAV2 ITR
     promoter        join(5487..5684,1..250)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 gcctggcggc gcgcgccacc gccgcctcag cactgaaggc gcgctgacgt cactcgccgg
       61 tcccccgcaa actccccttc ccggccacct tggtcgcgtc cgcgccgccg ccggcccagc
      121 cggaccgcac cacgcgaggc gcgagatagg ggggcacggg cgcgaccatc tgcgctgcgg
      181 cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg tgtcgtgcct
      241 gagagcgcag tcgaattcaa gctgctagca aggatccacc cgccaccatg ctgcagaacg
      301 agcttgctct taagttggct ggacttgata ttaacaagac tggaggaggt tctcatcatc
      361 atcatcatca tggtatggct agcatgactg gtggacagca aatgggtcgg gatctgtacg
      421 acgatgacga taaggatctc gcaacaatgg tcgactcatc acgtcgtaag tggaataagg
      481 caggtcacgc agtcagagct ataggtcggc tgagctcacc cgtggtttcc gagcggatgt
      541 accccgagga cggcgccctg aagagcgaga tcaagaaggg gctgaggctg aaggacggcg
      601 gccactacgc cgccgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
      661 gcgcctacat cgtggacatc aagttggaca tcgtgtccca caacgaggac tacaccatcg
      721 tggaacagtg cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
      781 agggaggtac aggcgggagt ctggtgagca agggcgagga ggataacatg gccatcatca
      841 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga
      901 tcgagggcga gggcgagggc cgcccctacg aggcctttca gaccgctaag ctgaaggtga
      961 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct
     1021 ccaaggccta cattaagcac ccagccgaca tccccgacta cttcaagctg tccttccccg
     1081 agggcttcag gtgggagcgc gtgatgaact tcgaggacgg cggcattatt cacgtgaacc
     1141 aggactcctc cctgcaggac ggcgtattca tctacaaggt gaagctgcgc ggcaccaact
     1201 tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggct acgcgtgacg
     1261 acctgactga agagcagatc gcagaattta aagaggcttt ctccctattt gacaaggacg
     1321 gggatgggac gataacaacc aaggagcttg ggacggtgtt ccgatcgctg gggcagaacc
     1381 ccacagaagc agagctgcag gacatgatca atgaagtaga tgccgacggt gacggcacat
     1441 tcgacttccc tgagttcctg acgatgatgg caagaaaaat gaatgacaca gacagtgaag
     1501 aggaaattcg cgaagcgttc cgcgtgtttg ataaggacgg caatggctac atcggcgcag
     1561 cagagcttcg ccacgtgatg acagaccttg gagagaagtt aacagatgag gaggttgatg
     1621 aaatgatcag ggtagcagac atcgatgggg atggtcaggt aaactacgaa gagtttgtac
     1681 aaatgatgac agcgaagtga aagcttatcg ataatcaacc tctggattac aaaatttgtg
     1741 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     1801 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     1861 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     1921 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     1981 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     2041 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     2101 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     2161 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     2221 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     2281 ccctttgggc cgcctccccg catcgatacc gtcgacctcg acccgggcgg ccgcttcgag
     2341 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     2401 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     2461 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt
     2521 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatcttc
     2581 ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc
     2641 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     2701 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     2761 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2821 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2881 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2941 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3001 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3061 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3121 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3181 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3241 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3301 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3361 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3421 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3481 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3541 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3601 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3661 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3721 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3781 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3841 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3901 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3961 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4021 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4081 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4141 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4201 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4261 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4321 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4381 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4441 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4501 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4561 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4621 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4681 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4741 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     4801 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     4861 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     4921 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     4981 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     5041 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     5101 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     5161 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
     5221 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctgcg
     5281 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
     5341 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
     5401 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
     5461 gatctctgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg
     5521 ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt
     5581 ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg
     5641 tgcgcactgc cagcttcagc accgcggaca gtgccttcgc cccc
//
