LOCUS       Exported                5563 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV vector expressing CaMPARI2_F391W (Kd = 110nM), a 
            photoconvertible fluorescent protein-based calcium integrator, in 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV_hsyn_NES-his-CaMPARI2-F391W-WPRE-SV40
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5563)
  AUTHORS   Moeyaert B, Holt G, Madangopal R, Perez-Alvarez A, Fearey BC, 
            Trojanowski NF, Ledderose J, Zolnik TA, Das A, Patel D, Brown TA, 
            Sachdev RNS, Eickholt BJ, Larkum ME, Turrigiano GG, Dana H, Gee CE, 
            Oertner TG, Hope BT, Schreiter ER
  TITLE     Improved methods for marking active neuron populations.
  JOURNAL   Nat Commun. 2018 Oct 25;9(1):4440. doi: 10.1038/s41467-018-06935-2.
  PUBMED    30361563
REFERENCE   2  (bases 1 to 5563)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5563
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        31..478
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             575..592
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             599..1039
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
     CDS             1760..1816
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHALRAIGRLSS"
     CDS             1835..1858
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1877..1903
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1922..1951
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1973..2561
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2026..2046)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2444..2455)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2564..2580)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2565..2581)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2610..2731
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2621..2640)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2675..2694
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2935..2952)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3106..3694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3186..3205)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3865..4725)
                     /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     4488..4507
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4726..4830)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4856..5311)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4993..5014)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5205..5224
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5383..5512
                     /label=AAV2 ITR
ORIGIN
        1 ggaagatctc tgcagagggc cctgcgtatg agtgcaagtg ggttttagga ccaggatgag
       61 gcggggtggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc acccaacccc
      121 cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga tgcggcgagg
      181 cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc
      241 gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc ccccgcaaac
      301 tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca
      361 cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg ccggcgactc
      421 agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga gagcgcagtc
      481 gaattcaagc tgctagcaag gatccgccac catgctgcag aacgagcttg ctcttaagtt
      541 ggctggactt gatattaaca agactggagg ttctcatcat caccaccacc atggatccga
      601 ccaactgact gaagagcaga tcgcagaatt taaagaggct ttctccctat ttgacaagga
      661 cggggatggg acaataacaa ccaaggagct ggggacggtg atgcggtctc tggggcagaa
      721 ccccacagaa gcagagctgc aggacatgat caatgaagta gatgccgacg gtgacggcac
      781 aatcgacttc cctgagttcc tgacaatgat ggcaagaaaa atgaaagaca cagacagtga
      841 agaagaaatt agagaagcgt tccgtgtgtt tgataaggat ggcaatggct acatcagtgc
      901 agcagagctt cgccacgtga tgacaaacct tggagagaag ttaacagatg aagaggttga
      961 tgaaatgatc agggaagcag acatcgatgg ggatggtcag gtaaactacg aagagtttgt
     1021 agtgatgatg acagcgaagc tcgagtgcga gaaaatctat gtgcgtgatg gagtgctgac
     1081 gggtgatatt catatgatct tgttgcttga aggaaatgcc cattaccgat gtgacttcag
     1141 aactacttac aaagctaagg agaagggtgt caagttacca ggcgtgcact atgtggacca
     1201 caccattgag attttaagcc atgacaaaga ttacaacaag gttaagatct atgagtatgc
     1261 tgttgctcat tctggattgc ctgacaatgc cagacgaggc ggtaccggcg gatccatggt
     1321 gagtgcgatt aagccagaca tgaagatcaa actgcgtatg gaaggcaacg taaacgggca
     1381 ccactttgtg atcgacggag atggtacagg caagccttat gagggaaaac agaccatgga
     1441 tcttgaagtc aaagagggcg gacctctgcc ttttgccttt gatatcctga ccactgcatt
     1501 ccattacggc aacagggtat tcgtgaaata tccagacaac atacaagact attttaagca
     1561 gtcgtttcct aaggggtatt cgtgggaacg aagcatgact ttcgaagacg ggggcatttg
     1621 ctatgccaga agcgacataa caatggaagg ggacactttc tataataaag ttcgatttta
     1681 tggtaccaac tttcccgcca atggtccagt tatgcagaag aagacgctga aatggatgcc
     1741 gagctggacg cgttcatcac gtcgtaagtg gaataagaca ggtcacgcac tgagagctat
     1801 aggtcggctg agctcaggag gttctggagg ttctgattac aaggacgacg atgacaaggg
     1861 aggttctgga ggttcttacc catacgacgt accagattac gctggaggtt ctggaggttc
     1921 tgagcagaag ctgatctcag aggaggacct gcgtacgtaa aagcttatcg ataatcaacc
     1981 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2041 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2101 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2161 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2221 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2281 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2341 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     2401 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     2461 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     2521 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg
     2581 acccgggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     2641 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     2701 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     2761 tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     2821 taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa
     2881 tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     2941 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3001 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3061 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3121 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3181 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3241 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3301 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3361 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3421 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3481 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3541 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3601 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3661 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3721 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3781 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3841 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3901 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3961 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4021 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4081 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4141 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4201 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4261 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4321 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4381 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4441 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4501 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4561 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4621 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4681 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4741 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4801 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     4861 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     4921 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     4981 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     5041 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     5101 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     5161 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     5221 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     5281 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     5341 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctgcgcgc tcgctcgctc
     5401 actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
     5461 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc cttgtagtta
     5521 atgattaacc cgccatgcta cttatctacg tagccatgct cta
//
