LOCUS       sequence-442661-        3558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses OCaMP, an orange calcium indicator, in bacteria. Contains 
            T7 promoter and RSET leader sequence peptide (His tag, T7 leader, 
            Xpress epitope).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3558)
  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 3558)
  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: pRSET OCaMP
FEATURES             Location/Qualifiers
     source          1..3558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             93..110
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             114..146
                     /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(150..158,159..173)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             189..245
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     941..960
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1490..1508)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1504..1551
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1568..1585)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1739..2327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1819..1838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2498..3289,3290..3358))
                     /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     3121..3140
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3359..3463)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3531..3549
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atgcggggtt ctcatcatca tcatcatcat ggtatggcta
      121 gcatgactgg tggacagcaa atgggtcggg atctgtacga cgatgacgat aaggatcgat
      181 ggggatcccg tcgtaagtgg aacaaggcag gtcacgcagt cagagctata ggtcggctga
      241 gctcacccgt ggtttccgag cggatgtacc ccgaggacgg cgtcctgaag agcgagatca
      301 agaaggggct gaggctgaag gacggcggcc actacgccgc cgaggtcaag accacctaca
      361 aggccaagaa gcccgtgcag ctgcccggcg cctacatcgt cgacataaag ttggacatcg
      421 tgtcccacaa cgaggactac accatcgtgg aacagtgcga acgcgccgag ggccgccacc
      481 ccaccggcgg cagggacgag ctgtacaagg gaggtacagg agggagtctg gtgagcaagg
      541 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
      601 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
      661 cctttcagac cgctaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
      721 tcctgtcccc tcagttcaca tacggctcca aggcctacat taagcaccca gccgacatcc
      781 ccgactactt taagctgtcc ttccccgagg gcttcaggtg ggagcgcgtg atgaacttcg
      841 aggacggcgg cattattcac gttaaccagg actcctccct gcaggacggc gtattcatct
      901 acaaggtgaa gctgcgcggc accaacttcc cccccgacgg ccccgtaatg cagaagaaga
      961 ccatgggctg ggaggctacg cgtgaccaac tgactgaaga gcagatcgca gaatttaaag
     1021 aggctttctc cctatttgac aaggacgggg atgggacgat aacaaccaag gagctgggga
     1081 cggtgttccg gtctctgggg cagaacccca cagaagcaga gctgcaggac atgatcaatg
     1141 aagtagatgc cgacggtgac ggcacattcg acttccctga gttcctgacg atgatggcaa
     1201 gaaagatgaa tgacacagac agtgaagaag aaattagaga agcgttccgc gtgtttgata
     1261 aggacggcaa tggctacatc ggcgcagcag agcttcgcca cgtgatgaca gaccttggag
     1321 agaagttaac agatgaggag gttgatgaaa tgatcagggt agcagacatc gatggggatg
     1381 gtcaggtaaa ctacgaagag tttgtacaaa tgatgacagc gaagtaagaa ttcgaagctt
     1441 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     1501 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gggcgctctt
     1561 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1621 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1681 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1741 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1801 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1861 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1921 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1981 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2041 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2101 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2161 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2221 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2281 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2341 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2401 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2461 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2521 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2581 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2641 atccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2701 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2761 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2821 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2881 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2941 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3001 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3061 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3121 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3181 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3241 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3301 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3361 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3421 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3481 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     3541 tcacgaggcc ctttcgtc
//
