LOCUS       sequence-237799-        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     128..147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     381..398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     483..502
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        483..501
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             533..555
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             575..592
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             596..628
                     /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(632..640,641..655)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1907..2614
                     /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(2680..2698)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2694..2741
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2838..3293
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2925..2944)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3135..3156
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3319..3423
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3424..3492,3493..4284)
                     /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(3642..3661)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4455..4816,1..227)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
       61 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
      301 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      361 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
      421 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg caggatctcg atcccgcgaa
      481 attaatacga ctcactatag ggagaccaca acggtttccc tctagaaata attttgttta
      541 actttaagaa ggagatatac atatgcgggg ttctcatcat catcatcatc atggtatggc
      601 tagcatgact ggtggacagc aaatgggtcg ggatctgtac gacgatgacg ataaggatcg
      661 atggggatcc atggtgagtg cgattaagcc agacatgaag atcaaactgc gtatggaagg
      721 caacgtaaac gggcaccact ttgtgatcga cggagatggt acaggcaagc cttatgaggg
      781 aaaacagacc atggatcttg aagtcaaaga gggcggacct ctgccttttg cctttgatat
      841 cctgaccact gcattcctgt acggcaacag ggtattcgtg aaatatccag acaacataca
      901 agactatttt aagcagtcgt ttcctaaggg gtattcgtgg gaacgaagca tgactttcga
      961 agacgggggc atttgctatg ccagaagcga cataacaatg gaaggggaca ctttctataa
     1021 taaagttcga ttttatggta ccaactttcc cgccaatggt ccagttatgc agaagaagac
     1081 gctgaaatgg gagccctcca ctgagaaaat gtatgtgcgt gatggagtgc tgacgggtga
     1141 tgtagagatg gctttgttgc ttgaaggaaa tgcccattac cgagcgttgt caagccgccg
     1201 caagttcaac aaaaccggcc atgccttgcg tgcaattggc cgcctgtcca gcggaggctc
     1261 tggcggttcc ggcggttcag gtggatcaga ccagctgacc gaggagcaaa ttgcagaatt
     1321 taaggaggcg ttctctttgt ttgacaaaga tggcgacggt acaattacaa ccaaggaatt
     1381 gggcacagta atgcgctccc ttggacagaa tcccactgaa gccgaacttc aagacatgat
     1441 caatgaggtt gatgctgacg gggatgggac tatcgacttt cctgagtttc ttaccatgat
     1501 ggctcgcaaa atgaaggata cagacagtga ggaggaaatt cgtgaggcct ttcgtgtgtt
     1561 tgataaggac gggaacggtt atatctcagc ggcggagctg cgtcacgtta tgaccaactt
     1621 aggcgaaaag ctgactgatg aagaagtcga tgagatgatc cgcgaggcgg acatcgatgg
     1681 tgatggtcaa gtcaattatg aagagtttgt ggtgatgatg accgcgaagt gcctcttcag
     1741 aactacttac aaagctaagg agaagggtgt caagttacca ggcgtgcact atgtggacca
     1801 caccattgag attttaagcc atgacaaaga ttacaacaag gttaagatct atgagtatgc
     1861 tgttgctcat tctggattgc ctgacaatgc cagacgagga ggtagcgtgt ctaagggcga
     1921 agagctgatc aaggaaaata tgcgtatgaa ggtggtcatg gaaggttcgg tcaacggcca
     1981 ccaattcaaa tgcacaggtg aaggagaagg cagaccgtac gagggagtgc aaaccatgag
     2041 gatcaaagtc atcgagggag gacccctgcc atttgccttt gacattcttg ccacgtcgtt
     2101 catgtatggc agccgtacct ttatcaagta cccggccgac atccctgatt tctttaaaca
     2161 gtcctttcct gagggtttta cttgggaaag agttacgaga tacgaagatg gtggagtcgt
     2221 caccgtcacg caggacacca gccttgagga tggcgagctc gtctacaacg tcaaggtcag
     2281 aggggtaaac tttccctcca atggtcccgt gatgcagaag aagaccaagg gttgggagcc
     2341 taatacagag atgatgtatc cagcagatgg tggtctgaga ggatacactg acatcgcact
     2401 gaaagttgat ggtggtggcc atctgcactg caacttcgtg acaacttaca ggtcaaaaaa
     2461 gaccgtcggg aacatcaaga tgcccggtgt ccatgccgtt gatcaccgcc tggaaaggat
     2521 cgaggagagt gacaatgaaa cctacgtagt gcaaagagaa gtggcagttg ccaaatacag
     2581 caaccttggt ggtggcatgg acgagctgta caagtaagaa ttcgaagctt gatccggctg
     2641 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     2701 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     2761 ccggatctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     2821 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2881 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2941 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3001 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3061 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3121 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3181 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3241 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     3301 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3361 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3421 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3481 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3541 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3601 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3661 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3721 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3781 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3841 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3901 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3961 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4021 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4081 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4141 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4201 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4261 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4321 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4381 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4441 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4501 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4561 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4621 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4681 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4741 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4801 ttggagcgaa cgacct
//
