LOCUS       Exported                5567 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV vector expressing CaMPARI2_H396K (Kd = 360nM), a 
            photoconvertible fluorescent protein-based calcium integrator, in 
            neurons.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV_hsyn_NES-his-CaMPARI2-H396K-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
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5567
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        30..477
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             574..591
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             598..1038
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
     CDS             1834..1857
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1876..1902
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1921..1950
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    1972..2560
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2025..2045)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2443..2454)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2563..2579)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2564..2580)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2609..2730
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2620..2639)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2674..2693
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3109..3697)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3189..3208)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3868..4728)
                     /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     4491..4510
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4729..4833)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4859..5314)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4996..5017)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5208..5227
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5386..5515
                     /label=AAV2 ITR
ORIGIN
        1 gaagatctct gcagagggcc ctgcgtatga gtgcaagtgg gttttaggac caggatgagg
       61 cggggtgggg gtgcctacct gacgaccgac cccgacccac tggacaagca cccaaccccc
      121 attccccaaa ttgcgcatcc cctatcagag agggggaggg gaaacaggat gcggcgaggc
      181 gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg
      241 cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact
      301 ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg accgcaccac
      361 gcgaggcgcg agataggggg gcacgggcgc gaccatctgc gctgcggcgc cggcgactca
      421 gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg
      481 aattcaagct gctagcaagg atccgccacc atgctgcaga acgagcttgc tcttaagttg
      541 gctggacttg atattaacaa gactggaggt tctcatcatc accaccacca tggatccgac
      601 caactgactg aagagcagat cgcagaattt aaagaggctt tctccctatt tgacaaggac
      661 ggggatggga caataacaac caaggagctg gggacggtga tgcggtctct ggggcagaac
      721 cccacagaag cagagctgca ggacatgatc aatgaagtag atgccgacgg tgacggcaca
      781 atcgacttcc ctgagttcct gacaatgatg gcaagaaaaa tgaaagacac agacagtgaa
      841 gaagaaatta gagaagcgtt ccgtgtgttt gataaggatg gcaatggcta catcagtgca
      901 gcagagcttc gccacgtgat gacaaacctt ggagagaagt taacagatga agaggttgat
      961 gaaatgatca gggaagcaga catcgatggg gatggtcagg taaactacga agagtttgta
     1021 gtgatgatga cagcgaagct cgagtgcgag aaaatctatg tgcgtgatgg agtgctgacg
     1081 ggtgatattc atatgatctt gttgcttgaa ggaaatgccc attaccgatg tgacttcaga
     1141 actacttaca aagctaagga gaagggtgtc aagttaccag gcgtgcacta tgtggaccac
     1201 accattgaga ttttaagcca tgacaaagat tacaacaagg ttaagatcta tgagtatgct
     1261 gttgctcatt ctggattgcc tgacaatgcc agacgaggcg gtaccggcgg atccatggtg
     1321 agtgcgatta agccagacat gaagatcaaa ctgcgtatgg aaggcaacgt aaacgggcac
     1381 cactttgtga tcgacggaga tggtacaggc aagccttatg agggaaaaca gaccatggat
     1441 cttgaagtca aagagggcgg acctctgcct tttgcctttg atatcctgac cactgcattc
     1501 cattacggca acagggtatt cgtgaaatat ccagacaaca tacaagacta ttttaagcag
     1561 tcgtttccta aggggtattc gtgggaacga agcatgactt tcgaagacgg gggcatttgc
     1621 tatgccagaa gcgacataac aatggaaggg gacactttct ataataaagt tcgattttat
     1681 ggtaccaact ttcccgccaa tggtccagtt atgcagaaga agacgctgaa atggatgccg
     1741 agctggacgc gttcatcacg tcgtaagttc aataagacag gtaaggcact gagagctata
     1801 ggtcggctga gctcaggagg ttctggaggt tctgattaca aggacgacga tgacaaggga
     1861 ggttctggag gttcttaccc atacgacgta ccagattacg ctggaggttc tggaggttct
     1921 gagcagaagc tgatctcaga ggaggacctg cgtacgtaaa agcttatcga taatcaacct
     1981 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2041 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2101 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2161 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2221 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     2281 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2341 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     2401 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     2461 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     2521 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga
     2581 cccgggcggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     2641 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2701 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2761 caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     2821 aaaatcgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat
     2881 cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     2941 gctcactgas kcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3001 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3061 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3121 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3181 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3241 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3301 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3361 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3421 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3481 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3541 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3601 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3661 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3721 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3781 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3841 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3901 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3961 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4021 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4081 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4141 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4201 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4261 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4321 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4381 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4441 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4501 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4561 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4621 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4681 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4741 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4801 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     4861 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4921 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     4981 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     5041 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     5101 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     5161 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     5221 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     5281 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
     5341 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctgcg cgctcgctcg
     5401 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
     5461 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag
     5521 ttaatgatta acccgccatg ctacttatct acgtagccat gctctag
//
