LOCUS       sequence-58020-e        4082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a single-wavelength red-shifted genetically encoded calcium 
            indicator constructed from calmodulin and cp-mRuby.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4082)
  AUTHORS   Akerboom J, Carreras Calderon N, Tian L, Wabnig S, Prigge M, Tolo J,
            Gordus A, Orger MB, Severi KE, Macklin JJ, Patel R, Pulver SR, 
            Wardill TJ, Fischer E, Schuler C, Chen TW, Sarkisyan KS, Marvin JS, 
            Bargmann CI, Kim DS, Kugler S, Lagnado L, Hegemann P, Gottschalk A, 
            Schreiter ER, Looger LL
  TITLE     Genetically encoded calcium indicators for multi-color neural 
            activity imaging and combination with optogenetics.
  JOURNAL   Front Mol Neurosci. 2013;6:2. doi: 10.3389/fnmol.2013.00002. Epub 
            2013 Mar 4.
  PUBMED    23459413
REFERENCE   2  (bases 1 to 4082)
  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: "Front Mol 
            Neurosci."; date: "2013"; volume: "6:2. doi"; pages: " 
            10.3389/fnmol.2013.00002. Epub 2013 Mar 4"
COMMENT     Sequence Label: pRSET-RCaMP1h
FEATURES             Location/Qualifiers
     source          1..4082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             109..126
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             130..162
                     /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(166..174,175..189)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      196..205
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             217..273
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     complement(1483..1501)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1497..1544
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1641..2096
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1728..1747)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1938..1959
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2122..2226
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2227..2295,2296..3087)
                     /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(2445..2464)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3258..3846
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3747..3766
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4000..4017
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgggttctca tcatcatcat
      121 catcatggta tggctagcat gactggtgga cagcaaatgg gtcgggatct gtacgacgat
      181 gacgataagg atctcgccac catggtcgac tcatcacgtc gtaagtggaa taagacaggt
      241 cacgcagtca gagctatagg tcggctgagc tcagcgatca acaccgaaat gatgtaccca
      301 gcggatggtg gtctgcgtgg ttacactcac atggcgctga aagttgatgg cggcggtcac
      361 ctgtcctgtt ctttcgtgac cacctaccgc tccaaaaaga ctgtcggcaa cattaagatg
      421 cctgccattc attacgtcag ccaccgtctg gagcgcctgg aggagagcga taacgaaatg
      481 tttgtcgtac agcgtgaaca cgcagttgcc aagtttgtgg gcctgggtgg tggcggcggt
      541 accggaggga gcatgaactc cctgatcaag gagaacatgc gtatgaaagt ggttctggaa
      601 ggctccgtaa acggccacca gttcaaatgc actggtgaag gcgaaggcaa cccgtatatg
      661 ggcacccaga ctatgcgtat caaagtgatc gagggtggtc cgctgccgtt tgcgttcgac
      721 atcctggcga cgtcctttat gtatggctcc cgtaccttca tcaaatatcc gaaaggcatc
      781 ccggatttct ttaagcagtc cttcccggaa ggttttacct gggaacgtgt gacccgttac
      841 gaagacggcg gcgtaattac cgttatgcaa gacacgtctc tggaggatgg ctgcctggtg
      901 tatcacgtgc aggttcgcgg tgttaacttc ccgagcaatg gtgctgtaat gcaaaagaaa
      961 accaaaggtt gggagcctac ggactcccaa ctgactgaag agcagatcgc agaatttaaa
     1021 gaggctttct ccctatttga caaggacggg gatgggacaa taacaaccaa ggagctgggg
     1081 acggtgatgc ggtctctggg gcagaacccc acagaagcag agctgcagga catgatcaat
     1141 gaagtagatg ccgacggtga cggcacaatc gacttccctg agttcctgat tatgatggca
     1201 agaaaaatga aatacacaga cagtgaagaa gaaattagag aagcgttcgg cgtgtttgat
     1261 aaggatggca atggctacat cagtgcagca gagcttcgcc acgtgatgac aaaccttgga
     1321 gagaagttaa cagatgaaga ggttgatgaa atgatcaggg aagcagacag cgatggggat
     1381 ggtcaggtaa actacgaaga gtttgtacaa atgatgacag cgaagtaaag cttgatccgg
     1441 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     1501 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     1561 tatccggatc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     1621 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1681 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     1741 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     1801 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     1861 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     1921 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     1981 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2041 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     2101 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2161 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     2221 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     2281 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     2341 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     2401 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     2461 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     2521 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     2581 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     2641 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     2701 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     2761 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     2821 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     2881 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     2941 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3001 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3061 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     3121 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     3181 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3241 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3301 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3361 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     3421 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3481 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3541 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3601 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3661 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3721 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3781 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3841 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3901 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3961 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     4021 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     4081 ag
//
