LOCUS       Exported                6343 bp DNA     circular SYN 20-JAN-2022
DEFINITION  AAV-mediated expression of ultrafast protein calcium sensor under 
            the CAG promoter, Cre-dependent expression.
ACCESSION   .
VERSION     .
KEYWORDS    pGP-AAV-CAG-FLEX-jGCaMP8m-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6343)
  AUTHORS   Zhang , Yan; Rozsa, Marton; Bushey, Daniel; Zheng, Jihong; Reep, 
            Daniel; Broussard, Gerard Joey; Tsang, Arthur; Tsegaye, Getahun; 
            Patel, Ronak; Narayan, Sujatha; Li, Jing Xuan; Zhang, Rongwei; 
            Ahrens, Misha B.; Turner, Glenn C.; Wang, Samuel S.-H.; Svoboda, 
            Karel; Korff, Wyatt; Schreiter, Eric R.; Hasseman, Jeremy P.; Kolb, 
            Ilya; Looger, Loren L
  TITLE     jGCaMP8 Fast Genetically Encoded Calcium Indicators
  JOURNAL   Janelia Research Campus. Online resource. 2020
REFERENCE   2  (bases 1 to 6343)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 4, 2022 from SnapGene 6.0.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Janelia 
            Research Campus. Online resource. 2020"
FEATURES             Location/Qualifiers
     source          1..6343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   45..174
                     /label=AAV2 ITR
     primer_bind     complement(56..67)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     117..128
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        485..762
                     /label=chicken beta-actin promoter
     primer_bind     1041..1063
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1125..1144
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1179..1212)
                     /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(1275..1308)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(1319..2635)
                     /codon_start=1
                     /label=NES-jGCaMP8m
                     /translation="MGNLNELALKLAGLDIMHHHHHHTRRKKTFKEVATAVKIIGRLMG
                     LKINVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGPVLLPDNHYLSVE
                     SKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEELFTGVVPILVEL
                     DGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDH
                     MKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNI
                     LGHKLEYNLPDQLTEEQIAEYKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQD
                     MINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGNGYISAAELRHVM
                     TNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     1878..1897
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2137..2158
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2264..2285)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(2567..2584)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2588..2629)
                     /codon_start=1
                     /label=NES
                     /label=jGCaMP8m (var 686)
                     /translation="NLNELALKLAGLDI"
     protein_bind    2652..2685
                     /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    2756..2789
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2831..3419
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2884..2904)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3302..3313)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3462..3583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3473..3492)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3527..3546
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3745..3874
                     /label=AAV2 ITR
     primer_bind     complement(3791..3802)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3852..3863
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3940..3957)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4111..4699)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4191..4210)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4870..5730)
                     /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     5493..5512
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5731..5835)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5861..6316)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5998..6019)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6210..6229
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc
       61 tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag
      121 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt
      181 taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag agtaccattg
      241 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      301 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      361 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      421 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      481 atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc
      541 ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg
      601 ggggggcgcg cgccgggggg gggggggggg gggggggggg ggggsggggs grggcggaga
      661 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
      721 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc
      781 cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc
      841 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc
      901 ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg
      961 gagggccctt tgtgcggggg gagcggctcg gggctgtccg cggggggacg gctgccttcg
     1021 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
     1081 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
     1141 tgtgctgtct catcattttg gcaaagaatt ggatccccat aacttcgtat aaagtatcct
     1201 atacgaagtt atatcaaaat aggaagacca atgcttcacc atcgacccga attgccaagc
     1261 atcaccatcg acccataact tcgtataatg tatgctatac gaagttatac tagagctttc
     1321 acttcgctgt catcatttgt acaaactctt cgtagtttac ctgaccatcc ccatcgatgt
     1381 ctgcttccct gatcatttca tcaacctctt catctgttaa cttctctcca aggtttgtca
     1441 tcacgtggcg aagctctgct gcactgatgt agccattgcc atccttatca aacacaccga
     1501 acgcttctct aatttcttct tccgtgtccc tgtatttcat ttttcttgcc atcattgtca
     1561 ggaactcagg gaagtcgatt gtgccgtcac cgtcggcatc tacttcattg atcatgtcct
     1621 gcagctctgc ttctgtgggg ttctgcccca gagaccgcat caccgtcccc agctccttgg
     1681 ttgttattgt cccatccccg tccttgtcaa atagggagaa agcctctttg tattctgcga
     1741 tctgctcttc agtcagttgg tccggcaggt tgtactccag cttgtgcccc aggatgttgc
     1801 cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga
     1861 acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct
     1921 ggatgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt
     1981 agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg
     2041 gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct
     2101 caccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga
     2161 tgggcaccac cccggtgaac agctcctcgc ccttgctcac catgctccct ccggtaccgc
     2221 ccttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     2281 ccatgtgatc gcgcttctcg ttggggtctt tcgaaagttt ggactccacg ctcaggtagt
     2341 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtagg
     2401 cgagctgcac gccgccgtcc tcgatgttgt ggcggatgtg gaagttcgcc ttgatgccgt
     2461 tcttctgctt gtcggccttg atatagacgt tgatcttcag gcccatcagg cggccgatga
     2521 tcttcacggc ggtggccacc tccttgaagg tcttcttgcg acgcgtgtga tgatggtgat
     2581 gatgcatgat gtccaaaccg gccaacttca atgccagttc gttcaggtta cccatggtgg
     2641 cggatcctag tataacttcg tataggatac tttatacgaa gttatcattg ggattcttcc
     2701 tattttgatc caagcatcac catcgaccct ctagtccaga tctcaccatc gacccataac
     2761 ttcgtatagc atacattata cgaagttatg tccctcgaag aggttcgaat tcgatatcaa
     2821 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2881 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2941 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     3001 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     3061 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3121 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3181 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     3241 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3301 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3361 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     3421 tcgataccgt cgacccgggc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     3481 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3541 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3601 tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct
     3661 ctacaaatgt ggtaaaatcg ataaggatct tcctagagca tggctacgta gataagtagc
     3721 atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc
     3781 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     3841 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcat taatgaatcg gccaacgcgc
     3901 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3961 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4021 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4081 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4141 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4201 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4261 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4321 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4381 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4441 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4501 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4561 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4621 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4681 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4741 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4801 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4861 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4921 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4981 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5041 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5101 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5161 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5221 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5281 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5341 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5401 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5461 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5521 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5581 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5641 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5701 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5761 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5821 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt
     5881 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     5941 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     6001 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     6061 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     6121 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     6181 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     6241 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     6301 ccgctacagg gcgcgtccca ttcgccattc aggctgcgca act
//
