LOCUS       sequence-313972-        3609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the pH-resistant long Stokes shift red fluorescent protein
            mCRISPRed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
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="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..604
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             608..640
                     /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(644..652,653..667)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      674..683
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1406..1423)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1473..1491)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1487..1534
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1631..2086
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1718..1737)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1928..1949
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2112..2216
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2217..2285,2286..3077)
                     /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(2435..2454)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3248..3609,1..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 ttctcatcac catcatcacc accatcatca
      601 ccacggtatg gctagcatga ctggtggaca gcaaatgggt cgggatctgt acgacgatga
      661 cgataaggat ccggccacca tggtgtctaa gggcgaagag ctgatcaagg aaaatatgcg
      721 tatgaaggtg gtcatggaag gttcggtcaa cggccaccaa ttcaaatgca caggtgaagg
      781 agaaggccgc ccgtacgaag gagtgcaaac catgaggatc aaagtcatcg agggaggacc
      841 cctgccattt gcctttgaca ttcttgccac gtcgttcatg tatggcagcc gtacttttat
      901 caagtacccg gccgatattc ctgatttctt taaacagtcc tttcctgagg gttttacttg
      961 ggaaagagtt acgagatacg aagatggtgg agtcgtcacc gtcacccagg acaccagcct
     1021 tgaggatggc gaactcgttt acaacgtcaa agtcagaggg gtaaactttc cctccaatgg
     1081 tcccgtgatg cagaagaaga ccaagggctg ggaggccgac acagagatga tgtatccagc
     1141 agatggtggt ctgaggggat accttgatcg ggcactgaaa gttgatggtg gtggccatct
     1201 gcattgcaac ttcgtaacaa cttacaggtc aaaaaagacc gtcggggaca tcaagatgcc
     1261 cggtgtccat gccgttgatc accgcctgga aaggatcgag gaaagtgaca atgaaaccta
     1321 cgtagtacaa cgcgaagtcg cagttgccaa gtacagcaac cttggtggtg ggatggacga
     1381 gctgtacaag tgataatagg aattcctata gtgtcaccta aatgatccgg ctgctaacaa
     1441 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     1501 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatc
     1561 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     1621 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1681 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     1741 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     1801 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     1861 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     1921 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     1981 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2041 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     2101 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2161 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2221 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2281 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2341 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2401 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     2461 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2521 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2581 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2641 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2701 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2761 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2821 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2881 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     2941 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3001 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3061 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3121 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3181 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3241 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3301 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3361 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3421 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3481 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3541 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3601 gaacgacct
//
