LOCUS       pAAV_hsyn_NES-his-CaMPARI2-F391W-G395D-WPRE-SV40                5567 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV vector expressing CaMPARI2_F391W-G395D (Kd = 530nM), a 
            photoconvertible fluorescent protein-based calcium integrator, in 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV_hsyn_NES-his-CaMPARI2-F391W-G395D-WPRE-SV40
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5567)
  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 5567)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            //gt12365.cn
FEATURES             Location/Qualifiers
     source          1..5567
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        20..467
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             564..581
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             588..1028
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
     CDS             1824..1847
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1866..1892
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1911..1940
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1962..2550
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2015..2035)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2433..2444)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2553..2569)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2554..2570)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2599..2720
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2610..2629)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2664..2683
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2928..2945)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3099..3687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3179..3198)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3858..4718)
                     /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     4481..4500
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4719..4823)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4849..5304)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4986..5007)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5198..5217
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5376..5505
                     /label=AAV2 ITR
ORIGIN
        1 gcagagggcc ctgcgtatga gtgcaagtgg gttttaggac caggatgagg cggggtgggg
       61 gtgcctacct gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa
      121 ttgcgcatcc cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac
      181 tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc
      241 gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg
      301 gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg accgcaccac gcgaggcgcg
      361 agataggggg gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc
      421 agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg aattcaagct
      481 gctagcaagg atccgccacc atgctgcaga acgagcttgc tcttaagttg gctggacttg
      541 atattaacaa gactggaggt tctcatcatc accaccacca tggatccgac caactgactg
      601 aagagcagat cgcagaattt aaagaggctt tctccctatt tgacaaggac ggggatggga
      661 caataacaac caaggagctg gggacggtga tgcggtctct ggggcagaac cccacagaag
      721 cagagctgca ggacatgatc aatgaagtag atgccgacgg tgacggcaca atcgacttcc
      781 ctgagttcct gacaatgatg gcaagaaaaa tgaaagacac agacagtgaa gaagaaatta
      841 gagaagcgtt ccgtgtgttt gataaggatg gcaatggcta catcagtgca gcagagcttc
      901 gccacgtgat gacaaacctt ggagagaagt taacagatga agaggttgat gaaatgatca
      961 gggaagcaga catcgatggg gatggtcagg taaactacga agagtttgta gtgatgatga
     1021 cagcgaagct cgagtgcgag aaaatctatg tgcgtgatgg agtgctgacg ggtgatattc
     1081 atatgatctt gttgcttgaa ggaaatgccc attaccgatg tgacttcaga actacttaca
     1141 aagctaagga gaagggtgtc aagttaccag gcgtgcacta tgtggaccac accattgaga
     1201 ttttaagcca tgacaaagat tacaacaagg ttaagatcta tgagtatgct gttgctcatt
     1261 ctggattgcc tgacaatgcc agacgaggcg gtaccggcgg atccatggtg agtgcgatta
     1321 agccagacat gaagatcaaa ctgcgtatgg aaggcaacgt aaacgggcac cactttgtga
     1381 tcgacggaga tggtacaggc aagccttatg agggaaaaca gaccatggat cttgaagtca
     1441 aagagggcgg acctctgcct tttgcctttg atatcctgac cactgcattc cattacggca
     1501 acagggtatt cgtgaaatat ccagacaaca tacaagacta ttttaagcag tcgtttccta
     1561 aggggtattc gtgggaacga agcatgactt tcgaagacgg gggcatttgc tatgccagaa
     1621 gcgacataac aatggaaggg gacactttct ataataaagt tcgattttat ggtaccaact
     1681 ttcccgccaa tggtccagtt atgcagaaga agacgctgaa atggatgccg agctggacgc
     1741 gttcatcacg tcgtaagtgg aataagacag atcacgcact gagagctata ggtcggctga
     1801 gctcaggagg ttctggaggt tctgattaca aggacgacga tgacaaggga ggttctggag
     1861 gttcttaccc atacgacgta ccagattacg ctggaggttc tggaggttct gagcagaagc
     1921 tgatctcaga ggaggacctg cgtacgtaaa agcttatcga taatcaacct ctggattaca
     1981 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     2041 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     2101 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     2161 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     2221 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     2281 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     2341 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     2401 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     2461 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     2521 gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga cccgggcggc
     2581 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     2641 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     2701 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     2761 gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata
     2821 aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac
     2881 aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgac
     2941 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3001 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3061 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3121 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3181 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3241 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3301 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3361 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3421 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3481 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3541 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3601 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3661 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3721 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3781 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3841 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3901 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3961 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4021 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4081 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4141 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4201 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4261 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4321 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4381 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4441 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4501 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4561 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4621 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4681 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4741 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4801 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     4861 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     4921 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     4981 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     5041 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     5101 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     5161 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     5221 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     5281 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     5341 ggcgatcggt gcgggcctct tcgctattac gccagctgcg cgctcgctcg ctcactgagg
     5401 ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc
     5461 gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag ttaatgatta
     5521 acccgccatg ctacttatct acgtagccat gctctaggaa gatctct
//