LOCUS       sequence-180853-        3571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of a fluorescent protein, monomeric Cherry 
            (mCherry).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3571)
  AUTHORS   Shaner NC, Campbell RE, Steinbach PA, Giepmans BN, Palmer AE, Tsien 
            RY
  TITLE     Improved monomeric red, orange and yellow fluorescent proteins 
            derived from Discosoma sp. red fluorescent protein.
  JOURNAL   Nat Biotechnol. 2004 Dec;22(12):1567-72. Epub 2004 Nov 21.
  PUBMED    15558047
REFERENCE   2  (bases 1 to 3571)
  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: "Nat 
            Biotechnol."; date: "2004-12"; volume: "22(12)"; pages: "1567-72. 
            Epub 2004 Nov 21"
COMMENT     Sequence Label: pNCS mCherry
FEATURES             Location/Qualifiers
     source          1..3571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     65..84
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        65..83
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             115..137
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             157..174
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             178..210
                     /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(214..222,223..237)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      244..253
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             250..954
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGRPYEGTQ
                     TAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFED
                     GGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGE
                     IKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEG
                     RHSTGGMDELYK"
     primer_bind     complement(389..407)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     657..676
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1017..1035)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1031..1078
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1175..1630
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1262..1281)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1472..1493
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1656..1760
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1761..1829,1830..2621)
                     /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(1979..1998)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2792..3380
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3281..3300
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3534..3551
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaggatct cgatcccgcg
       61 aaattaatac gactcactat agggagacca caacggtttc cctctagaaa taattttgtt
      121 taactttaag aaggagatat acatatgcgg ggttctcatc atcatcatca tcatggtatg
      181 gctagcatga ctggtggaca gcaaatgggt cgggatctgt acgacgatga cgataaggat
      241 cccgccacca tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
      301 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
      361 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
      421 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
      481 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
      541 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
      601 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
      661 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
      721 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
      781 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
      841 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
      901 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaagaattc
      961 gaagcttgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     1021 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     1081 gaaaggagga actatatccg gatctggcgt aatagcgaag aggcccgcac cgatcgccct
     1141 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     1201 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     1261 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     1321 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     1381 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     1441 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     1501 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     1561 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     1621 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1681 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1741 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1801 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1861 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1921 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1981 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     2041 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     2101 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     2161 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2221 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2281 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     2341 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     2401 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     2461 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2521 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2581 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2641 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2701 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2761 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2821 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2881 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     2941 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3001 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3061 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3121 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3181 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3241 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3301 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3361 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3421 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3481 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3541 cagtgagcga ggaagcggaa gagcgcccaa t
//
