LOCUS       sequence-216914-        3577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For bacterial expression and purification of mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3577)
  AUTHORS   DeGroot ACM, Busch DJ, Hayden CC, Mihelic SA, Alpar AT, Behar M, 
            Stachowiak JC
  TITLE     Entropic Control of Receptor Recycling Using Engineered Ligands.
  JOURNAL   Biophys J. 2018 Mar 27;114(6):1377-1388. doi: 
            10.1016/j.bpj.2018.01.036.
  PUBMED    29590595
REFERENCE   2  (bases 1 to 3577)
  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.bpj.2018.01"; journalName: "Biophys J"; date: 
            "2018-03-27- 27"; volume: "114"; issue: "6"; pages: "1377-1388"
COMMENT     Sequence Label: pRSET his-mCherry
FEATURES             Location/Qualifiers
     source          1..3577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     62..81
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        62..80
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             112..134
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             154..171
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             175..207
                     /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(211..219,220..234)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             250..957
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(392..410)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     660..679
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1020..1038)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1034..1081
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1178..1633
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1265..1284)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1475..1496
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1659..1763
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1764..1832,1833..2624)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPKERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1982..2001)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2795..3383
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3284..3303
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3537..3554
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 caaaccgcct ctccccgcgc gttggccgat tcattaatgc aggatctcga tcccgcgaaa
       61 ttaatacgac tcactatagg gagaccacaa cggtttccct ctagaaataa ttttgtttaa
      121 ctttaagaag gagatataca tatgcggggt tctcatcatc atcatcatca tggtatggct
      181 agcatgactg gtggacagca aatgggtcgg gatctgtacg acgatgacga taaggatcga
      241 tggggatccg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
      301 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
      361 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
      421 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
      481 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
      541 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctc
      601 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
      661 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
      721 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
      781 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
      841 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
      901 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaagaa
      961 ttcgaagctt gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     1021 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1081 gctgaaagga ggaactatat ccggatctgg cgtaatagcg aagaggcccg caccgatcgc
     1141 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     1201 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     1261 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     1321 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     1381 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     1441 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     1501 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     1561 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     1621 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1681 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1741 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     1801 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     1861 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     1921 agatccttga gagttttcgc cccaaagaac gttttccaat gatgagcact tttaaagttc
     1981 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     2041 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2101 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2161 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2221 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2281 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     2341 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     2401 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2461 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     2521 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2581 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     2641 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     2701 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2761 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2821 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2881 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2941 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3001 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3061 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3121 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3181 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3241 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3301 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3361 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3421 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3481 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3541 agtcagtgag cgaggaagcg gaagagcgcc caatacg
//
