LOCUS       sequence-264435-        5090 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses red fluorescent calcium sensor in hippocampal slices.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5090)
  AUTHORS   Kerruth S, Coates C, Durst CD, Oertner TG, Torok K
  TITLE     The kinetic mechanisms of fast-decay red-fluorescent genetically 
            encoded calcium indicators.
  JOURNAL   J Biol Chem. 2019 Mar 15;294(11):3934-3946. doi: 
            10.1074/jbc.RA118.004543. Epub 2019 Jan 16.
  PUBMED    30651353
REFERENCE   2  (bases 1 to 5090)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1074/jbc.RA118.004543"; journalName: "J Biol Chem"; date: 
            "2019-03-15- 15"; volume: "294"; issue: "11"; pages: "3934-3946"
COMMENT     Sequence Label: pCI-syn-fRCaMP1
FEATURES             Location/Qualifiers
     source          1..5090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        92..539
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     intron          576..708
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     691..710
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        753..771
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             928..945
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             949..981
                     /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(985..993,994..1008)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1036..1092
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKYNKTGHAVRAIGRLSS"
     polyA_signal    2271..2392
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2282..2301)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2336..2355
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2573..3028
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2660..2679)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2870..2891
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3095..3114)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3214..3236
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3274..3292)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3360..3464
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3465..3533,3534..4325)
                     /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(3683..3702)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4496..5084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4985..5004
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catggctcga
       61 cagatctaat ctgcagaggg ccctgcgtat gagtgcaagt gggttttaag accaggatga
      121 ggcggggtgg gggtgcctac ctgacgaccg accccgaccc actggacaag cacccaaccc
      181 ccattcccca aattgcgcat cccctatcag agagggggag gggaaacagg atgcggcgag
      241 gcgcgtgcgc actgccagct tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg
      301 cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc actcgccggt cccccgcaaa
      361 ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc
      421 acgcgaggcg cgagataggg gggcacgggc gcgaccatct gcgctgcggc gccggcgact
      481 cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagc
      541 tgcagctagg aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt acaagacagg
      601 tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat
      661 aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt gtccactccc
      721 agttcaatta cagctcttaa ggctagagta cttaatacga ctcactatag gctagcgcta
      781 ccggactcaa gcttggtacc gaactcggat ccactagtaa cggccgccag tgtgctggaa
      841 ttcggcttgg ggatatccac catggtgcag aacgagcttg ctcttaagtt ggctggactt
      901 gatattaaca agactggagg aggttctcat catcatcatc atcatggtat ggctagcatg
      961 actggtggac agcaaatggg tcgggatctg tacgacgatg acgataagga tctcgcaaca
     1021 atggtcgact catcacgtcg taagtacaat aagacaggtc acgcagtcag agctataggt
     1081 cggctgagct cagcgatcaa ctgcgaaatg atgtacccag cggatggtgg tctgcgtggt
     1141 tacactcaca tggcgctgaa agttgatggc ggcggtcacc tgtcctgttc tttcgtgacc
     1201 acctaccgct ccaaaaagac tgtcggcaac attaagatgc ctgccattca tagcgtcagc
     1261 caccgtctgg agcgcctgga ggagagcgat aacgaaatgt ttgtcgtaca gcgtgaacac
     1321 gcagttgcca agtttgtggg cctgggtggt ggcggcggta ccggagggag catgaactcc
     1381 ctgatcaagg agaacatgcg tatgaaagtg gttctggaag gctccgtaaa cggccaccag
     1441 ttcaaatgca ctggtgaagg cgaaggcaac ccgtatatgg gcacccagac tatgcgtatc
     1501 aaagtgatcg agggtggtcc gctgccgttt gcgttcgaca tcctggcgac gtcctttatg
     1561 tatggctccc gtaccttcat caaatatccg aaaggcatcc cggatttctt taagcagtcc
     1621 ttcccggaag gttttacctg ggaacgtgtg acccgttacg aagacggcgg cgtaattacc
     1681 gttatgcaag acacgtctct ggaggatggc tgcctggtgt atcacgtgca ggttcgcggt
     1741 gtgaacttcc cgagcaatgg tgctgtaatg caaaagaaaa ccaaaggttg ggagcctacg
     1801 gactcccaaa ccactgaaga gcagatcgca gaatttaaag aggctttctc cctatttgac
     1861 aaggacgggg atgggacaat aacaaccaag gagctgggga cggtgatgcg gtctctgggg
     1921 cagaacccca cagaagcaga gctgcaggac atgatcaatg aagtagatgc cgacggtgac
     1981 ggcacaatcg acttccctga gttcctgatt atgatggcaa gaaaaatgaa atacacagac
     2041 agtgaagaag aaattagaga agcgttcggc gtgtttgata aggatggcaa tggctacatc
     2101 agtgcagcag agcttcgcca cgtgatgaca aaccttggag agaagttaac agatgaagag
     2161 gttgatgaaa tgatcaggga agcagacagc gatggggatg gtcaggtaaa ctacgaagag
     2221 tttgtacaaa tgatgacagc gaagtaggcg gccgcttcga gcagacatga taagatacat
     2281 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2341 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2401 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa
     2461 gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg ggctggcgta atagcgaaga
     2521 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc
     2581 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2641 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2701 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2761 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2821 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2881 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2941 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3001 tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     3061 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     3121 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     3181 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     3241 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     3301 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     3361 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3421 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3481 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3541 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     3601 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     3661 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3721 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3781 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3841 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3901 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3961 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     4021 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     4081 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     4141 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     4201 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4261 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4321 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     4381 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4441 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4501 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4561 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4621 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4681 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4741 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4801 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4861 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4921 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4981 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5041 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
//
