LOCUS       Exported                5657 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV expression of jRCaMP1a, a red fluorescent calcium sensor 
            protein, from the Synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.Syn.NES.jRCaMP1a.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"
     CDS             174..191
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             195..227
                     /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             231..254
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             282..338
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     misc_feature    1506..2094
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1559..1579)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1977..1988)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2097..2113)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2098..2114)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2143..2264
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2154..2173)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2208..2227
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2426..2555
                     /label=AAV2 ITR
     primer_bind     complement(2621..2638)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2792..3380)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2872..2891)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3551..4411)
                     /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     4174..4193
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4412..4516)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4542..4997)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4679..4700)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4891..4910
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5069..5198
                     /label=AAV2 ITR
     promoter        5280..70
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg tgtcgtgcct
       61 gagagcgcag tcgaattcaa gctgctagca aggatccacc cgccaccatg ctgcagaacg
      121 agcttgctct taagttggct ggacttgata ttaacaagac tggaggaggt tctcatcatc
      181 atcatcatca tggtatggct agcatgactg gtggacagca aatgggtcgg gatctgtacg
      241 acgatgacga taaggatctc gcaacaatgg tcgactcatc acgtcgtaag tggaataaga
      301 caggtcacgc agtcagagct ataggtcggc tgagctcagc gatcaactgc gaaatgatgt
      361 acccagcgga tggtggtctg cgtggttaca ctcacatggc gctgaaagtt gatggcggcg
      421 gtcacctgtc ctgttctttc gtgaccacct accgctccaa aaagactgtc ggcaacatta
      481 agatgcctgc cattcatagc gtcagccacc gtctggagcg cctggaggag agcgataacg
      541 aaatgtttgt cgtacagcgt gaacacgcag ttgccaagtt tgtgggcctg ggtggtggcg
      601 gcggtaccgg agggagcatg aactccctga tcaaggagaa catgcgtatg aaagtggttc
      661 tggaaggctc cgtaaacggc caccagttca aatgcactgg tgaaggcgaa ggcaacccgt
      721 atatgggcac ccagactatg cgtatcaaag tgatcgaggg tggtccgctg ccgtttgcgt
      781 tcgacatcct ggcgacgtcc tttatgtatg gctcccgtac cttcatcaaa tatccgaaag
      841 gcatcccgga tttctttaag cagtccttcc cggaaggttt tacctgggaa cgtgtgaccc
      901 gttacgaaga cggcggcgta attaccgtta tgcaagacac gtctctggag gatggctgcc
      961 tggtgtatca cgtgcaggtt cgcggtgtga acttcccgag caatggtgct gtaatgcaaa
     1021 agaaaaccaa aggttgggag cctacggact cccaaaccac tgaagagcag atcgcagaat
     1081 ttaaagaggc tttctcccta tttgacaagg acggggatgg gacaataaca accaaggagc
     1141 tggggacggt gatgcggtct ctggggcaga accccacaga agcagagctg caggacatga
     1201 tcaatgaagt agatgccgac ggtgacggca caatcgactt ccctgagttc ctgattatga
     1261 tggcaagaaa aatgaaatac acagacagtg aagaagaaat tagagaagcg ttcggcgtgt
     1321 ttgataagga tggcaatggc tacatcagtg cagcagagct tcgccacgtg atgacaaacc
     1381 ttggagagaa gttaacagat gaagaggttg atgaaatgat cagggaagca gacagcgatg
     1441 gggatggtca ggtaaactac gaagagtttg tacaaatgat gacagcgaag tgaaagctta
     1501 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     1561 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     1621 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     1681 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     1741 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     1801 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     1861 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     1921 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     1981 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     2041 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     2101 accgtcgacc tcgacccggg cggccgcttc gagcagacat gataagatac attgatgagt
     2161 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     2221 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     2281 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc
     2341 tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt agataagtag
     2401 catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct
     2461 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     2521 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc ggccaacgcg
     2581 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2641 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2701 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2761 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2821 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2881 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2941 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3001 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3061 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3121 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3181 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3241 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3301 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3361 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3421 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3481 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3541 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3601 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3661 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3721 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3781 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3841 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3901 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3961 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4021 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4081 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4141 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4201 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4261 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4321 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4381 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4441 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4501 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     4561 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4621 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4681 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4741 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4801 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4861 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4921 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4981 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     5041 ggtgcgggcc tcttcgctat tacgccagct gcgcgctcgc tcgctcactg aggccgcccg
     5101 ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
     5161 cagagaggga gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc
     5221 atgctactta tctacgtagc catgctctag gaagatctct gcagagggcc ctgcgtatga
     5281 gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac
     5341 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
     5401 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
     5461 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
     5521 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
     5581 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
     5641 gaccatctgc gctgcgg
//
