LOCUS       Exported                6373 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV expression of CaMPARI, a photoconvertible fluorescent 
            protein-based calcium integrator.
ACCESSION   .
VERSION     .
KEYWORDS    pENN.AAV.CAG.Flex.CaMPARI.WPRE.SV40
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6373)
  AUTHORS   Fosque BF, Sun Y, Dana H, Yang CT, Ohyama T, Tadross MR, Patel R, 
            Zlatic M, Kim DS, Ahrens MB, Jayaraman V, Looger LL, Schreiter ER
  TITLE     Neural circuits. Labeling of active neural circuits in vivo with 
            designed calcium integrators.
  JOURNAL   Science. 2015 Feb 13;347(6223):755-60. doi: 10.1126/science.1260922.
  PUBMED    25678659
REFERENCE   2  (bases 1 to 6373)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6373
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   2..131
                     /label=AAV2 ITR
     promoter        442..719
                     /label=chicken beta-actin promoter
     primer_bind     998..1020
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1082..1101
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1136..1169)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    complement(1232..1265)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             1378..1395
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1402..1842
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             2563..2619
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     protein_bind    2639..2672
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    2743..2776
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2818..3406
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2871..2891)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3289..3300)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3449..3570
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3460..3479)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3514..3533
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3732..3861
                     /label=AAV2 ITR
     primer_bind     complement(3927..3944)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4098..4686)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4178..4197)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4857..5717)
                     /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     5480..5499
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5718..5822)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5848..6303)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5985..6006)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6197..6216
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
       61 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      121 taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt agccatgctc
      181 taggaagagt accattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      241 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      301 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      361 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      421 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      481 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      541 tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg
      601 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      661 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      721 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      781 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      841 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      901 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ctgtccgcgg
      961 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1021 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1081 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgga tccccataac
     1141 ttcgtataaa gtatcctata cgaagttata tcaaaatagg aagaccaatg cttcaccatc
     1201 gacccgaatt gccaagcatc accatcgacc cataacttcg tataatgtat gctatacgaa
     1261 gttatactag ggatcccccc tcgaaattca agctgctagc aaggatccgc caccatgctg
     1321 cagaacgagc ttgctcttaa gttggctgga cttgatatta acaagactgg aggttctcat
     1381 catcaccacc accatggatc cgaccaactg actgaagagc agatcgcaga atttaaagag
     1441 gctttctccc tatttgacaa ggacggggat gggacaataa caaccaagga gctggggacg
     1501 gtgatgcggt ctctggggca gaaccccaca gaagcagagc tgcaggacat gatcaatgaa
     1561 gtagatgccg acggtgacgg cacaatcgac ttccctgagt tcctgacaat gatggcaaga
     1621 aaaatgaaag acacagacag tgaagaagaa attagagaag cgttccgtgt gtttgataag
     1681 gatggcaatg gctacatcag tgcagcagag cttcgccacg tgatgacaaa ccttggagag
     1741 aagttaacag atgaagaggt tgatgaaatg atcagggaag cagacatcga tggggatggt
     1801 caggtaaact acgaagagtt tgtacaaatg atgacagcga agctcgagtg cgagaaaatc
     1861 tatgtgcgtg atggagtgct gacgggtgat attcatatga tcttgttgct tgaaggaaat
     1921 gcccattacc gatgtgactt cagaactact tacaaagcta aggagaaggg tgtcaagtta
     1981 ccaggcgtgc actttgtgga ccactgcatt gagattttaa gccatgacaa agattacaac
     2041 aaggttaagc tgtatgagta tgctgttgct cattctggat tgcctgacaa tgccagacga
     2101 ggcggtaccg gcggatccat ggtgagtgcg attaagccag acatgaagat caaactccgt
     2161 atggaaggca acgtaaacgg gcaccacttt gtgatcgacg gagatggtac aggcaagcct
     2221 tatgagggaa aacagaccat ggatcttgaa gtcaaagagg gcggacctct gccttttgcc
     2281 tttgatatcc tgaccactgc attccattac ggcaacaggg tattcgtgaa atatccagac
     2341 aacatacaag actattttaa gcagtcgttt cctaaggggt attcgtggga acgaagcatg
     2401 actttcgaag acgggggcat ttgctatgcc agaaacgaca taacaatgga aggggacact
     2461 ttctataata aagttcgatt ttatggtacc aactttcccg ccaatggtcc agttatgcag
     2521 aagaagacgc tgaaatggat gccgagctgg acgcgttcat cacgtcgtaa gtggaataag
     2581 acaggtcacg cagtcagagc tataggtcgg ctgagctcat aaaagctcga attctagtat
     2641 aacttcgtat aggatacttt atacgaagtt atcattggga ttcttcctat tttgatccaa
     2701 gcatcaccat cgaccctcta gtccagatct caccatcgac ccataacttc gtatagcata
     2761 cattatacga agttatgtcc ctcgaagagg ttcgaattcg atatcaagct tatcgataat
     2821 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     2881 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     2941 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     3001 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3061 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3121 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3181 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     3241 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3301 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3361 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga
     3421 cccgggcggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     3481 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     3541 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     3601 caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     3661 aaaatcgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat
     3721 cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     3781 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     3841 agtgagcgag cgagcgcgca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3901 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3961 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4021 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4081 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4141 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4201 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4261 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4321 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4381 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4441 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4501 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4561 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4621 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4681 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4741 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4801 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4861 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4921 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4981 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5041 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5101 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5161 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5221 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5281 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5341 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5401 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5461 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5521 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5581 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5641 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5701 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5761 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5821 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     5881 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     5941 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     6001 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     6061 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     6121 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     6181 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     6241 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     6301 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     6361 cgctattacg cca
//
