LOCUS       sequence-178047-        4775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Long stokes-shift variant of red fluorescent protein mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4775)
  AUTHORS   Shen Y, Chen Y, Wu J, Shaner NC, Campbell RE
  TITLE     Engineering of mCherry variants with long Stokes shift, red-shifted 
            fluorescence, and low cytotoxicity.
  JOURNAL   PLoS One. 2017 Feb 27;12(2):e0171257. doi: 
            10.1371/journal.pone.0171257. eCollection 2017.
  PUBMED    28241009
REFERENCE   2  (bases 1 to 4775)
  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: "PLoS One.";
            date: "2017-02-27"; volume: "12(2):e0171257. doi"; pages: " 
            10.1371/journal.pone.0171257. eCollection 2017"
COMMENT     Sequence Label: pBAD-LSSmCherry1
FEATURES             Location/Qualifiers
     source          1..4775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             352..384
                     /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(388..396,397..411)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             427..1137
                     /codon_start=1
                     /product="derivative of the monomeric red fluorescent 
                     protein mCherry"
                     /label=mCherry2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMTIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFNWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTMGLEASTERMYPEDGALK
                     GESKERLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLDILSHNEDYTIVEQYERS
                     EGRHSTGGMDELYK"
     primer_bind     complement(572..590)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(1176..1193)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1346..1432
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1524..1551
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1570..1661
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1662..1730,1731..2522)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALITFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1880..1899)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2693..3281
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3182..3201
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3435..3452
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3467..3607
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3533..3552)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3693..3715
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3871..4749)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggggggttct catcatcatc atcatcatgg tatggctagc
      361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc
      421 tcgagaatgg tgagcaaggg cgaggaggat aacatgacca tcatcaagga gttcatgcgc
      481 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
      541 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
      601 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
      661 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaattgg
      721 gagcgcgtga tgaactttga ggacggcggt gtggtgaccg tgacccagga ctcctccctg
      781 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctctgacggc
      841 cccgtaatgc agtgtcgtac catgggcttg gaggccagta ctgagcggat gtaccccgag
      901 gacggcgccc tgaagggcga gagtaaggag aggctgaagc tgaaggacgg cggccactac
      961 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     1021 aacgtcgaca tcaagttgga catcctttcc cacaacgagg actacaccat cgtggaacag
     1081 tacgaacgct ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaaaag
     1141 cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc
     1201 agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac
     1261 cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat
     1321 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     1381 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg
     1441 agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata
     1501 aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct
     1561 acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1621 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     1681 gtgtcgccct tattaccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     1741 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     1801 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     1861 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     1921 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     1981 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2041 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2101 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2161 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2221 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2281 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2341 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2401 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2461 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2521 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     2581 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     2641 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2701 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2761 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2821 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2881 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2941 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3001 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3061 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3121 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3181 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3241 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3301 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3361 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3421 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     3481 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
     3541 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     3601 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3661 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3721 accgtcatca ccgaaacgcg cgaggcagca gatcaattcg cgcgcgaagg cgaagcggca
     3781 tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat gctactccgt
     3841 caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta catcattcac
     3901 tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt ttttaaatac
     3961 ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg cgataggcat
     4021 ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa
     4081 gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac
     4141 atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg
     4201 acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa tcgcttccat
     4261 gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat agcgcccttc
     4321 cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc
     4381 atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc attcatgcca
     4441 gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct ccggatgacg
     4501 accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa
     4561 ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga gaatataacc
     4621 tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct caatcggcgt
     4681 taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat cattttgcgc
     4741 ttcagccata cttttcatac tcccgccatt cagag
//
