LOCUS       Exported                5624 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV expression of ultrasensitive protein calcium sensor from 
            Synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.Syn.GCaMP6f.WPRE.SV40
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5624)
  AUTHORS   Chen TW, Wardill TJ, Sun Y, Pulver SR, Renninger SL, Baohan A, 
            Schreiter ER, Kerr RA, Orger MB, Jayaraman V, Looger LL, Svoboda K, 
            Kim DS
  TITLE     Ultrasensitive fluorescent proteins for imaging neuronal activity.
  JOURNAL   Nature. 2013 Jul 18;499(7458):295-300. doi: 10.1038/nature12354.
  PUBMED    23868258
REFERENCE   2  (bases 1 to 5624)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5624
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     regulatory      258..267
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             264..1616
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      360..369
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     650..671
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(777..798)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1038..1057)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    1629..2217
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1682..1702)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2100..2111)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2220..2236)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2221..2237)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2266..2387
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2277..2296)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2331..2350
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2549..2678
                     /label=AAV2 ITR
     primer_bind     complement(2744..2761)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2915..3503)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2995..3014)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3674..4534)
                     /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     4297..4316
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4535..4639)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4665..5120)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4802..4823)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5014..5033
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5192..5321
                     /label=AAV2 ITR
     promoter        5403..226
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 cctcagcact gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc cccttcccgg
       61 ccaccttggt cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga
      121 gatagggggg cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag cgctgcctca
      181 gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga attcaagctg
      241 ctagcaagga tccacccgcc accatgggtt ctcatcatca tcatcatcat ggtatggcta
      301 gcatgactgg tggacagcaa atgggtcggg atctgtacga cgatgacgat aaggatctcg
      361 ccaccatggt cgactcatca cgtcgtaagt ggaataagac aggtcacgca gtcagagcta
      421 taggtcggct gagctcactc gagaacgtct atatcaaggc cgacaagcag aagaacggca
      481 tcaaggcgaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgcctacc
      541 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
      601 tgagcgtgca gtccaaactt tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc
      661 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggcggta
      721 ccggagggag catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      781 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggtgagggcg
      841 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
      901 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
      961 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac atccaggagc
     1021 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1081 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1141 tcctggggca caagctggag tacaacctgc cggaccaact gactgaagag cagatcgcag
     1201 aatttaaaga ggaattctcc ctatttgaca aggacgggga tgggacaata acaaccaagg
     1261 agctggggac ggtgatgcgg tctctggggc agaaccccac agaagcagag ctgcaggaca
     1321 tgatcaatga agtagatgcc gacggtgacg gcacaatcga cttccctgag ttcctgacaa
     1381 tgatggcaag aaaaatgaaa tacagggaca cggaagaaga aattagagaa gcgttcggtg
     1441 tgtttgataa ggatggcaat ggctacatca gtgcagcaga gcttcgccac gtgatgacaa
     1501 accttggaga gaagttaaca gatgaagagg ttgatgaaat gatcagggaa gcagacatcg
     1561 atggggatgg tcaggtaaac tacgaagagt ttgtacaaat gatgacagcg aagtgaaagc
     1621 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     1681 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     1741 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     1801 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     1861 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     1921 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     1981 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     2041 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     2101 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     2161 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     2221 gataccgtcg acctcgaccc gggcggccgc ttcgagcaga catgataaga tacattgatg
     2281 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     2341 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     2401 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa
     2461 acctctacaa atgtggtaaa atcgataagg atcttcctag agcatggcta cgtagataag
     2521 tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc
     2581 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     2641 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcattaatga atcggccaac
     2701 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2761 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2821 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2881 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2941 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3001 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3061 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3121 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3181 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3241 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3301 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3361 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3421 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3481 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3541 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3601 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3661 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3721 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3781 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3841 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3901 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3961 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4021 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4081 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4141 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4201 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4261 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4321 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4381 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4441 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4501 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4561 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4621 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
     4681 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4741 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     4801 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     4861 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     4921 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     4981 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     5041 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     5101 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
     5161 atcggtgcgg gcctcttcgc tattacgcca gctgcgcgct cgctcgctca ctgaggccgc
     5221 ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
     5281 gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa tgattaaccc
     5341 gccatgctac ttatctacgt agccatgctc taggaagatc tctgcagagg gccctgcgta
     5401 tgagtgcaag tgggttttag gaccaggatg aggcggggtg ggggtgccta cctgacgacc
     5461 gaccccgacc cactggacaa gcacccaacc cccattcccc aaattgcgca tcccctatca
     5521 gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc ttcagcaccg
     5581 cggacagtgc cttcgccccc gcctggcggc gcgcgccacc gccg
//
