LOCUS Exported 4082 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION a single-wavelength red-shifted genetically encoded calcium indicator constructed from calmodulin and cp-mRuby. ACCESSION . VERSION . KEYWORDS pRSET-RCaMP1h 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4082 /organism="synthetic DNA construct" /mol_type="other DNA" 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 166..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 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" 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 2227..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 //