LOCUS       sequence-442110-        5799 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pAAV (non-flex) for expressing OCaMP, an orange calcium indicator, 
            in mammalian cells and in vivo. Contains CMV promoter and RSET 
            leader sequence peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5799)
  AUTHORS   Aggarwal A, Baker HA, Durst CD, Chen I-W, de Chambrier P, Gonzales 
            JM, Marvin JS, Vandal M, Lundberg T, Sakoi K, Patel RH, Wang C-Y, 
            Visser F, Fouad Y, Sunil S, Wiens M, Terai T, Takahashi-Yamashiro K,
            Thompson RJ, Brown TA, Nasu Y, Nguyen MD, Gordon GRJ, McFarlane S, 
            Podgorski K, Holtmaat A, Campbell RE, Lohman AW
  TITLE     A sensitive orange fluorescent calcium ion indicator for imaging 
            neural activity
  JOURNAL   bioRxiv 2025.07.28.667269; doi: 
            https://doi.org/10.1101/2025.07.28.667269
REFERENCE   2  (bases 1 to 5799)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2025.07.28.667269; doi: https://doi.org/10.1101/2025.07.28.667269"
COMMENT     Sequence Label: pAAV-CMV-OCaMP-WPRE
FEATURES             Location/Qualifiers
     source          1..5799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     protein_bind    137..157
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     enhancer        218..521
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        522..725
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     676..696
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    746..770
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             843..860
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             864..896
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(900..908,909..923)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             939..995
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     1691..1710
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2178..2202)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2212..2800
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2265..2285)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2683..2694)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2830..2847)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2836..3043
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3051..3191
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3051..3180
                     /label=AAV2 ITR
     rep_origin      3266..3721
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3353..3372)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3563..3584
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3738..3757)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3857..3879
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3917..3935)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4003..4107
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4108..4176,4177..4968)
                     /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     complement(4326..4345)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5139..5727
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5628..5647
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5789..5799,1..130)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tctagacaac tttgtataga aaagttgtag ttattaatag taatcaatta
      181 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      241 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      301 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      361 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      421 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      481 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      541 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      601 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      661 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      721 gagctggttt agtgaaccgt cagatccaag tttgtacaaa aaagcaggct gccaccatgc
      781 tgcagaacga gcttgctctt aagttggctg gacttgatat taacaagact ggaggaggtt
      841 ctcatcatca tcatcatcat ggtatggcta gcatgactgg tggacagcaa atgggtcggg
      901 atctgtacga cgatgacgat aaggatcgat ggggatcccg tcgtaagtgg aacaaggcag
      961 gtcacgcagt cagagctata ggtcggctga gctcacccgt ggtttccgag cggatgtacc
     1021 ccgaggacgg cgtcctgaag agcgagatca agaaggggct gaggctgaag gacggcggcc
     1081 actacgccgc cgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     1141 cctacatcgt cgacataaag ttggacatcg tgtcccacaa cgaggactac accatcgtgg
     1201 aacagtgcga acgcgccgag ggccgccacc ccaccggcgg cagggacgag ctgtacaagg
     1261 gaggtacagg agggagtctg gtgagcaagg gcgaggagga taacatggcc atcatcaagg
     1321 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
     1381 agggcgaggg cgagggccgc ccctacgagg cctttcagac cgctaagctg aaggtgacca
     1441 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcaca tacggctcca
     1501 aggcctacat taagcaccca gccgacatcc ccgactactt taagctgtcc ttccccgagg
     1561 gcttcaggtg ggagcgcgtg atgaacttcg aggacggcgg cattattcac gttaaccagg
     1621 actcctccct gcaggacggc gtattcatct acaaggtgaa gctgcgcggc accaacttcc
     1681 cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggctacg cgtgaccaac
     1741 tgactgaaga gcagatcgca gaatttaaag aggctttctc cctatttgac aaggacgggg
     1801 atgggacgat aacaaccaag gagctgggga cggtgttccg gtctctgggg cagaacccca
     1861 cagaagcaga gctgcaggac atgatcaatg aagtagatgc cgacggtgac ggcacattcg
     1921 acttccctga gttcctgacg atgatggcaa gaaagatgaa tgacacagac agtgaagaag
     1981 aaattagaga agcgttccgc gtgtttgata aggacggcaa tggctacatc ggcgcagcag
     2041 agcttcgcca cgtgatgaca gaccttggag agaagttaac agatgaggag gttgatgaaa
     2101 tgatcagggt agcagacatc gatggggatg gtcaggtaaa ctacgaagag tttgtacaaa
     2161 tgatgacagc gaagtaaacc cagctttctt gtacaaagtg ggaattccga taatcaacct
     2221 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2281 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2341 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2401 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2461 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     2521 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2581 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     2641 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     2701 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     2761 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgggaatt cctagagctc
     2821 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2881 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2941 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     3001 gcaaggggga ggattgggaa gagaatagca ggcatgctgg ggagggccgc aggaacccct
     3061 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     3121 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     3181 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3241 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3301 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     3361 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3421 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3481 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3541 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     3601 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     3661 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     3721 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     3781 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3841 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3901 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3961 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4021 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4081 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4141 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4201 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4261 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4321 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4381 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4441 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4501 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4561 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4621 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4681 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4741 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4801 aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt
     4861 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4921 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4981 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5041 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5101 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5161 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5221 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5281 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5341 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5401 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5461 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5521 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5581 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5641 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5701 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5761 gccttttgct ggccttttgc tcacatgtcc tgcaggcag
//
