LOCUS       sequence-308219-        3609 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses the pH-resistant long Stokes shift red fluorescent protein
            mCRISPRed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3609)
  AUTHORS   Erdogan M, Fabritius A, Basquin J, Griesbeck O
  TITLE     Targeted In Situ Protein Diversification and Intra-organelle 
            Validation in Mammalian Cells.
  JOURNAL   Cell Chem Biol. 2020 May 21;27(5):610-621.e5. doi: 
            10.1016/j.chembiol.2020.02.004. Epub 2020 Mar 5.
  PUBMED    32142629
REFERENCE   2  (bases 1 to 3609)
  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"; doi: 
            "10.1016/j.chembiol.2020.02.004"; journalName: "Cell Chem Biol"; 
            date: "2020-05-21- 21"; volume: "27"; issue: "5"; pages: 
            "610-621.e5"
COMMENT     Sequence Label: mCRISPRed pRSETB
FEATURES             Location/Qualifiers
     source          1..3609
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     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..141
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             145..177
                     /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(181..189,190..204)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      211..220
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(943..960)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1010..1028)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1024..1071
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1168..1623
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1255..1274)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1465..1486
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1649..1753
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1754..1822,1823..2614)
                     /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(1972..1991)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2785..3373
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3274..3293
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3527..3544
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcaccat
      121 catcaccacc atcatcacca cggtatggct agcatgactg gtggacagca aatgggtcgg
      181 gatctgtacg acgatgacga taaggatccg gccaccatgg tgtctaaggg cgaagagctg
      241 atcaaggaaa atatgcgtat gaaggtggtc atggaaggtt cggtcaacgg ccaccaattc
      301 aaatgcacag gtgaaggaga aggccgcccg tacgaaggag tgcaaaccat gaggatcaaa
      361 gtcatcgagg gaggacccct gccatttgcc tttgacattc ttgccacgtc gttcatgtat
      421 ggcagccgta cttttatcaa gtacccggcc gatattcctg atttctttaa acagtccttt
      481 cctgagggtt ttacttggga aagagttacg agatacgaag atggtggagt cgtcaccgtc
      541 acccaggaca ccagccttga ggatggcgaa ctcgtttaca acgtcaaagt cagaggggta
      601 aactttccct ccaatggtcc cgtgatgcag aagaagacca agggctggga ggccgacaca
      661 gagatgatgt atccagcaga tggtggtctg aggggatacc ttgatcgggc actgaaagtt
      721 gatggtggtg gccatctgca ttgcaacttc gtaacaactt acaggtcaaa aaagaccgtc
      781 ggggacatca agatgcccgg tgtccatgcc gttgatcacc gcctggaaag gatcgaggaa
      841 agtgacaatg aaacctacgt agtacaacgc gaagtcgcag ttgccaagta cagcaacctt
      901 ggtggtggga tggacgagct gtacaagtga taataggaat tcctatagtg tcacctaaat
      961 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     1021 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
     1081 ggaactatat ccggatctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     1141 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     1201 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     1261 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     1321 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     1381 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     1441 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     1501 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     1561 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     1621 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1681 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1741 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1801 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1861 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1921 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1981 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     2041 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     2101 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     2161 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     2221 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     2281 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     2341 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2401 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2461 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2521 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2581 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2641 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2701 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2761 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2821 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2881 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2941 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3001 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3061 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3121 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3181 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3241 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3301 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3361 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3421 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3481 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3541 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     3601 ttaatgcag
//
