LOCUS       Exported                5684 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV expression of jRGECO1a, a red fluorescent calcium sensor 
            protein, from the Synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.Syn.NES-jRGECO1a.WPRE.SV40
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 May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5684
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        211..658
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             762..779
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             783..815
                     /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             819..842
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             870..926
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     1622..1641
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2121..2709
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2174..2194)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2592..2603)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2712..2728)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2713..2729)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2758..2879
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2769..2788)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2823..2842
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3041..3170
                     /label=AAV2 ITR
     rep_origin      3242..3697
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3329..3348)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3539..3560
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3723..3827
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3828..4688
                     /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(4046..4065)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4859..5447
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5348..5367
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5601..5618
                     /label=L4440
                     /note="L4440 vector, forward primer"
     repeat_region   5684..129
                     /label=AAV2 ITR
ORIGIN
        1 tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg
       61 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
      121 ggggttcctt gtagttaatg attaacccgc catgctactt atctacgtag ccatgctcta
      181 ggaagatctc tgcagagggc cctgcgtatg agtgcaagtg ggttttagga ccaggatgag
      241 gcggggtggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc acccaacccc
      301 cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga tgcggcgagg
      361 cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc
      421 gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc ccccgcaaac
      481 tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca
      541 cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg ccggcgactc
      601 agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga gagcgcagtc
      661 gaattcaagc tgctagcaag gatccacccg ccaccatgct gcagaacgag cttgctctta
      721 agttggctgg acttgatatt aacaagactg gaggaggttc tcatcatcat catcatcatg
      781 gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac gatgacgata
      841 aggatctcgc aacaatggtc gactcatcac gtcgtaagtg gaataaggca ggtcacgcag
      901 tcagagctat aggtcggctg agctcacccg tggtttccga gcggatgtac cccgaggacg
      961 gcgccctgaa gagcgagatc aagaaggggc tgaggctgaa ggacggcggc cactacgccg
     1021 ccgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacatcg
     1081 tggacatcaa gttggacatc gtgtcccaca acgaggacta caccatcgtg gaacagtgcg
     1141 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag ggaggtacag
     1201 gcgggagtct ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     1261 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     1321 gcgagggccg cccctacgag gcctttcaga ccgctaagct gaaggtgacc aagggtggcc
     1381 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctaca
     1441 ttaagcaccc agccgacatc cccgactact tcaagctgtc cttccccgag ggcttcaggt
     1501 gggagcgcgt gatgaacttc gaggacggcg gcattattca cgtgaaccag gactcctccc
     1561 tgcaggacgg cgtattcatc tacaaggtga agctgcgcgg caccaacttc ccccccgacg
     1621 gccccgtaat gcagaagaag accatgggct gggaggctac gcgtgacgac ctgactgaag
     1681 agcagatcgc agaatttaaa gaggctttct ccctatttga caaggacggg gatgggacga
     1741 taacaaccaa ggagcttggg acggtgttcc gatcgctggg gcagaacccc acagaagcag
     1801 agctgcagga catgatcaat gaagtagatg ccgacggtga cggcacattc gacttccctg
     1861 agttcctgac gatgatggca agaaaaatga atgacacaga cagtgaagag gaaattcgcg
     1921 aagcgttccg cgtgtttgat aaggacggca atggctacat cggcgcagca gagcttcgcc
     1981 acgtgatgac agaccttgga gagaagttaa cagatgagga ggttgatgaa atgatcaggg
     2041 tagcagacat cgatggggat ggtcaggtaa actacgaaga gtttgtacaa atgatgacag
     2101 cgaagtgaaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     2161 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     2221 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     2281 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     2341 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     2401 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     2461 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     2521 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     2581 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     2641 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     2701 cctccccgca tcgataccgt cgacctcgac ccgggcggcc gcttcgagca gacatgataa
     2761 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     2821 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     2881 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt
     2941 aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatcttcct agagcatggc
     3001 tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag
     3061 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     3121 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctggcgtaat
     3181 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     3241 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3301 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3361 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3421 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3481 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3541 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3601 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3661 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     3721 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3781 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3841 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3901 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3961 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4021 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4081 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4141 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4201 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4261 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4321 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4381 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4441 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4501 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4561 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4621 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4681 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4741 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4801 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4861 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4921 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4981 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5041 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5101 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5161 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5221 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5281 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5341 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5401 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5461 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5521 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5581 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     5641 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagc
//
