LOCUS       sequence-232846-        4816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses rsCaMPARI in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4816)
  AUTHORS   Sha F, Abdelfattah AS, Patel R, Schreiter ER
  TITLE     Erasable labeling of neuronal activity using a reversible calcium 
            marker.
  JOURNAL   Elife. 2020 Sep 15;9. pii: 57249. doi: 10.7554/eLife.57249.
  PUBMED    32931424
REFERENCE   2  (bases 1 to 4816)
  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: "Elife."; 
            date: "2020-09-15"; volume: "9. pii: 57249. doi"; pages: " 
            10.7554/eLife.57249"
COMMENT     Sequence Label: pRSET_His-rsCaMPARI-mRuby3
FEATURES             Location/Qualifiers
     source          1..4816
                     /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             112..129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             133..165
                     /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(169..177,178..192)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1444..2151
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTDI
                     ALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAVA
                     KYSNLGGGMDELYK"
     primer_bind     complement(2217..2235)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2231..2278
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2375..2830
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2462..2481)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2672..2693
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2856..2960
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2961..3029,3030..3821)
                     /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(3179..3198)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3992..4580
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4481..4500
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4734..4751
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
      181 gatgacgata aggatcgatg gggatccatg gtgagtgcga ttaagccaga catgaagatc
      241 aaactgcgta tggaaggcaa cgtaaacggg caccactttg tgatcgacgg agatggtaca
      301 ggcaagcctt atgagggaaa acagaccatg gatcttgaag tcaaagaggg cggacctctg
      361 ccttttgcct ttgatatcct gaccactgca ttcctgtacg gcaacagggt attcgtgaaa
      421 tatccagaca acatacaaga ctattttaag cagtcgtttc ctaaggggta ttcgtgggaa
      481 cgaagcatga ctttcgaaga cgggggcatt tgctatgcca gaagcgacat aacaatggaa
      541 ggggacactt tctataataa agttcgattt tatggtacca actttcccgc caatggtcca
      601 gttatgcaga agaagacgct gaaatgggag ccctccactg agaaaatgta tgtgcgtgat
      661 ggagtgctga cgggtgatgt agagatggct ttgttgcttg aaggaaatgc ccattaccga
      721 gcgttgtcaa gccgccgcaa gttcaacaaa accggccatg ccttgcgtgc aattggccgc
      781 ctgtccagcg gaggctctgg cggttccggc ggttcaggtg gatcagacca gctgaccgag
      841 gagcaaattg cagaatttaa ggaggcgttc tctttgtttg acaaagatgg cgacggtaca
      901 attacaacca aggaattggg cacagtaatg cgctcccttg gacagaatcc cactgaagcc
      961 gaacttcaag acatgatcaa tgaggttgat gctgacgggg atgggactat cgactttcct
     1021 gagtttctta ccatgatggc tcgcaaaatg aaggatacag acagtgagga ggaaattcgt
     1081 gaggcctttc gtgtgtttga taaggacggg aacggttata tctcagcggc ggagctgcgt
     1141 cacgttatga ccaacttagg cgaaaagctg actgatgaag aagtcgatga gatgatccgc
     1201 gaggcggaca tcgatggtga tggtcaagtc aattatgaag agtttgtggt gatgatgacc
     1261 gcgaagtgcc tcttcagaac tacttacaaa gctaaggaga agggtgtcaa gttaccaggc
     1321 gtgcactatg tggaccacac cattgagatt ttaagccatg acaaagatta caacaaggtt
     1381 aagatctatg agtatgctgt tgctcattct ggattgcctg acaatgccag acgaggaggt
     1441 agcgtgtcta agggcgaaga gctgatcaag gaaaatatgc gtatgaaggt ggtcatggaa
     1501 ggttcggtca acggccacca attcaaatgc acaggtgaag gagaaggcag accgtacgag
     1561 ggagtgcaaa ccatgaggat caaagtcatc gagggaggac ccctgccatt tgcctttgac
     1621 attcttgcca cgtcgttcat gtatggcagc cgtaccttta tcaagtaccc ggccgacatc
     1681 cctgatttct ttaaacagtc ctttcctgag ggttttactt gggaaagagt tacgagatac
     1741 gaagatggtg gagtcgtcac cgtcacgcag gacaccagcc ttgaggatgg cgagctcgtc
     1801 tacaacgtca aggtcagagg ggtaaacttt ccctccaatg gtcccgtgat gcagaagaag
     1861 accaagggtt gggagcctaa tacagagatg atgtatccag cagatggtgg tctgagagga
     1921 tacactgaca tcgcactgaa agttgatggt ggtggccatc tgcactgcaa cttcgtgaca
     1981 acttacaggt caaaaaagac cgtcgggaac atcaagatgc ccggtgtcca tgccgttgat
     2041 caccgcctgg aaaggatcga ggagagtgac aatgaaacct acgtagtgca aagagaagtg
     2101 gcagttgcca aatacagcaa ccttggtggt ggcatggacg agctgtacaa gtaagaattc
     2161 gaagcttgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     2221 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     2281 gaaaggagga actatatccg gatctggcgt aatagcgaag aggcccgcac cgatcgccct
     2341 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     2401 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2461 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2521 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2581 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2641 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2701 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2761 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2821 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2881 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2941 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3001 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3061 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3121 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3181 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3241 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3301 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3361 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3421 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3481 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     3541 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3601 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3661 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3721 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3781 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3841 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3901 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3961 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4021 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4081 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4141 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4201 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4261 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4321 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4381 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4441 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4501 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4561 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4621 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4681 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     4741 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     4801 cgattcatta atgcag
//
