LOCUS       sequence-314708-        6296 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AAV-mediated expression of ultrafast protein calcium sensor under 
            the CAG promoter, Cre-dependent expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6296)
  AUTHORS   Zhang Y, Rozsa M, Bushey D, Zheng J, Reep D, Broussard GJ, Tsang A, 
            Tsegaye G, Patel R, Narayan S, Li JX, Zhang R, Ahrens MB, Turner GC,
            Wang SS-H, Svoboda K, Korff W, Schreiter ER, Hasseman JP, Kolb I, 
            Looger LL
  TITLE     jGCaMP8 Fast Genetically Encoded Calcium Indicators
  JOURNAL   Janelia Research Campus, Online Resource (2020) 
            doi:10.25378/janelia.13148243.
REFERENCE   2  (bases 1 to 6296)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Janelia 
            Research Campus, Online Resource (2020) 
            doi:10.25378/janelia.13148243."
COMMENT     Sequence Label: pGP-AAV-CAG-FLEX-jGCaMP8s-WPRE
FEATURES             Location/Qualifiers
     source          1..6296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   45..174
                     /label=AAV2 ITR
     promoter        485..762
                     /label=chicken beta-actin promoter
     primer_bind     1042..1064
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1126..1145
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1180..1213
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    1276..1309
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1879..1898
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2138..2159
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2265..2286)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(2568..2585)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(2605..2638)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(2709..2742)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2784..3372
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2837..2857)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3255..3266)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3415..3536
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3426..3445)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3480..3499
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3698..3827
                     /label=AAV2 ITR
     rep_origin      complement(4064..4652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4144..4163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4823..5614,5615..5683))
                     /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     5446..5465
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5684..5788)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5814..6269)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5951..5972)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6163..6182
                     /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 cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga
      661 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
      721 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgacgctg
      781 ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac
      841 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg
      901 cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg
      961 ggagggccct ttgtgcgggg ggagcggctc ggggctgtcc gcggggggac ggctgccttc
     1021 gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct
     1081 ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta
     1141 ttgtgctgtc tcatcatttt ggcaaagaat tggatcccca taacttcgta taaagtatcc
     1201 tatacgaagt tatatcaaaa taggaagacc aatgcttcac catcgacccg aattgccaag
     1261 catcaccatc gacccataac ttcgtataat gtatgctata cgaagttata ctagagcttt
     1321 cacttcgctg tcatcatttg tacaaactct tcgtagttta cctgaccatc cccatcgatg
     1381 tctgcttccc tgatcatttc atcaacctct tcatctgtta acttctctcc aaggtttgtc
     1441 atcacgtggc gaagctctgc tgcactgatg tagccattgc catccttatc aaacacaccg
     1501 aacgcttctc taatttcttc ttccgtgtcc ctgtatttca tttttcttgc catcattgtc
     1561 aggaactcag ggaagtcgat tgtgccgtca ccgtcggcat ctacttcatt gatcatgtcc
     1621 tgcagctctg cttctgtggg gttgtgtccg agagaccgca tcaccgtccc cagctccttg
     1681 gttgttattg tcccatcccc gtccttgtca aatagggaga aagcctcttt gtattctgcg
     1741 atctgctctt cagtcagttg gtccggcagg ttgtactcca gcttgtgccc caggatgttg
     1801 ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg
     1861 aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc
     1921 tggatgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg
     1981 tagcggctga agcactgcac gccgtaggtc agggtggtca cgagggtggg ccagggcacg
     2041 ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc
     2101 tcaccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg
     2161 atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatgctccc tccggtaccg
     2221 cccttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg
     2281 accatgtgat cgcgcttctc gttggggtct ttcgaaagtt tggactccac gctcaggtag
     2341 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtag
     2401 gcgagctgca cgccgccgtc ctcgatgttg tggcggatgt ggaagttcgc cttgatgccg
     2461 ttcttctgct tgtcggcctt gatatagacg ttgatcttca ggcccatcag catggcgatg
     2521 atcttcacgg cggtggccac ctccttgaag gtcttcttgc gacgcgtgtg atgatggtga
     2581 tgatgcatgg tggcggatcc tagtataact tcgtatagga tactttatac gaagttatca
     2641 ttgggattct tcctattttg atccaagcat caccatcgac cctctagtcc agatctcacc
     2701 atcgacccat aacttcgtat agcatacatt atacgaagtt atgtccctcg aagaggttcg
     2761 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     2821 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     2881 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     2941 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     3001 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     3061 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     3121 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     3181 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     3241 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     3301 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     3361 ccgcctcccc gcatcgatac cgtcgacccg ggcggccgct tcgagcagac atgataagat
     3421 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     3481 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     3541 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
     3601 gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tcttcctaga gcatggctac
     3661 gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg
     3721 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     3781 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cattaatgaa
     3841 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3901 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3961 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4021 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4081 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4141 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4201 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4261 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4321 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4381 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4441 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4501 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4561 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4621 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4681 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4741 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4801 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4861 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4921 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4981 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5041 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5101 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5161 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5221 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5281 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5341 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5401 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5461 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5521 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5581 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5641 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5701 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5761 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     5821 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     5881 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     5941 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     6001 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     6061 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     6121 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     6181 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     6241 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaact
//
