LOCUS       Exported                5657 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV expression of jRCaMP1b, a red fluorescent calcium sensor 
            protein, from the Synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.Syn.NES-jRCaMP1b.WPRE.SV40
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5657)
  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 5657)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5657
                     /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"
     misc_feature    2094..2682
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2147..2167)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2565..2576)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2685..2701)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2686..2702)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2731..2852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2742..2761)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2796..2815
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3014..3143
                     /label=AAV2 ITR
     rep_origin      3215..3670
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3302..3321)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3512..3533
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3696..3800
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3801..4661
                     /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(4019..4038)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4832..5420
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5321..5340
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5574..5591
                     /label=L4440
                     /note="L4440 vector, forward primer"
     repeat_region   5657..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 gactcatcgc gacgtaagtg gaataagtgg ggtcacgcag
      901 tcagagctat aggtcggctg agctcagcga acaacaccga aatgatgtac ccagcggatg
      961 gtggtctgcg tggttacact cacatggcgc tgaaagttga tggcggcggt cacctgtcct
     1021 gttctttcgt gaccacctac cgctccaaaa agactgtcgg caacattaag atgcctgcca
     1081 ttcattacgt cagccaccgt ctggagcgcc tggaggagag cgataacgaa atgtttgtcg
     1141 tacagcgtga acacgcagtt gccaagtttg tgggcctggg tggtggcggc ggtaccggag
     1201 ggagcatgaa ctccctgatc aaggagaaca tgcgtatgaa agtggttctg gaaggctccg
     1261 taaacggcca ccagttcaaa tgcactggtg aaggcgaagg caacccgtat atgggcaccc
     1321 agactatgcg tatcaaagtg atcgagggtg gtccgctgcc gtttgcgttc gacatcctgg
     1381 cgacgtcctt tatgtatggc tcccgtacct tcatcaaata tccgaaaggc atcccggatt
     1441 tctttaagca gtccttcccg gaaggtttta cctgggaacg tgtgacccgt tacgaagacg
     1501 gcggcgtaat taccgttatg caagacacgt ctctggagga tggctgcctg gtgtatcacg
     1561 tgcaggttcg cggtgtgaac ttcccgagca atggtgctgt aatgcaaaag aaaaccaaag
     1621 gttgggagcc tacggactcc caactgactg aagagcagat cgcagaattt aaagaggctt
     1681 tctccctatt tgacaaggac ggggatggga caataacaac caaggagatg gggacggtga
     1741 tgcggtctct ggggcagaac cccacagaag cagagctgca ggacatgatc aatgaagtag
     1801 atgccgacgg tgacggcaca atcgacttcc ctgagttcct gattatgatg gcaggcaaaa
     1861 tgaaatacac agacagtgaa gaagaaatta gagaagcgtt cggcgtgttt gataaggatg
     1921 gcaatggcta catcagtgca gcagagcttc gccacgtgat gacaaacctt ggagagaagt
     1981 taacagatga agaggttgat gaaatgatca gggaagcaga cagcgatggg gatggtcagg
     2041 taaactacga agagtttgta caaatgatga cagcgaagtg aaagcttatc gataatcaac
     2101 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2161 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     2221 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     2281 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     2341 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     2401 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2461 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     2521 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     2581 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     2641 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc
     2701 gacccgggcg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     2761 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2821 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     2881 ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     2941 gtaaaatcga taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta
     3001 atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3061 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3121 tcagtgagcg agcgagcgcg cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     3181 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     3241 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3301 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3361 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3421 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3481 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3541 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3601 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     3661 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3721 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3781 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3841 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3901 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3961 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4021 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4081 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4141 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4201 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4261 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4321 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     4381 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4441 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4501 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4561 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4621 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4681 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4741 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4801 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4861 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4921 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4981 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5041 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5101 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5161 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5221 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5281 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5341 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5401 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     5461 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5521 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     5581 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     5641 cgattcatta atgcagc
//
