LOCUS       sequence-265067-        3627 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Constitutive expression of mRuby3 in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3627)
  AUTHORS   Bajar BT, Wang ES, Lam AJ, Kim BB, Jacobs CL, Howe ES, Davidson MW, 
            Lin MZ, Chu J
  TITLE     Improving brightness and photostability of green and red fluorescent
            proteins for live cell imaging and FRET reporting.
  JOURNAL   Sci Rep. 2016 Feb 16;6:20889. doi: 10.1038/srep20889.
  PUBMED    26879144
REFERENCE   2  (bases 1 to 3627)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2016-02-16"; volume: "6:20889. doi"; pages: " 
            10.1038/srep20889"
COMMENT     Sequence Label: pNCS-mRuby3
FEATURES             Location/Qualifiers
     source          1..3627
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             122..144
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             164..181
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             185..217
                     /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(221..229,230..244)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             251..961
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     primer_bind     complement(1028..1046)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1042..1089
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1186..1641
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1273..1292)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1483..1504
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1667..1771
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1772..1840,1841..2632)
                     /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(1990..2009)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2803..3391
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3292..3311
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3545..3562
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaatcaccg agtttattct tgacacctga tgcgatgaat gatataatag gaaagtactg
      121 ttttgtttaa ctttaagaag gagatataca tatgcggggt tctcatcatc atcatcatca
      181 tggtatggct agcatgactg gtggacagca aatgggtcgg gatctgtacg acgatgacga
      241 taaggatccg atggtgtcta agggcgaaga gctgatcaag gaaaatatgc gtatgaaggt
      301 ggtcatggaa ggttcggtca acggccacca attcaaatgc acaggtgaag gagaaggcag
      361 accgtacgag ggaactcaaa ccatgaggat caaagtcatc gagggaggac ccctgccatt
      421 tgcctttgac attcttgcca cgtcgttcat gtatggcagc cgtactttta tcaagtaccc
      481 ggccgacatc cctgatttct ttaaacagtc ctttcctgag ggttttactt gggaaagagt
      541 tacgagatac gaagatggtg gagtcgtcac cgtcacgcag gacaccagcc ttgaggatgg
      601 cgagctcgtc tacaacgtca aggtcagagg ggtaaacttt ccctccaatg gtcccgtgat
      661 gcagaagaag accaagggtt gggagcctaa tacagagatg atgtatccag cagatggtgg
      721 tctgagagga tacactgaca tcgcactgaa agttgatggt ggtggccatc tgcactgcaa
      781 cttcgtgaca acttacaggt caaaaaagac cgtcgggaac atcaagatgc ccggtgtcca
      841 tgccgttgat caccgcctgg aaaggatcga ggagagtgac aatgaaacct acgtagtgca
      901 acgcgaagtg gcagttgcca aatacagcaa ccttggtggt ggcatggacg agctgtacaa
      961 gtaatgaatt cgaagcttga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     1021 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     1081 ggttttttgc tgaaaggagg aactatatcc ggatctggcg taatagcgaa gaggcccgca
     1141 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     1201 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1261 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     1321 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     1381 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     1441 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     1501 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     1561 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     1621 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1681 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     1741 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     1801 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     1861 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     1921 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     1981 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     2041 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2101 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2161 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2221 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2281 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     2341 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     2401 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2461 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2521 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2581 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     2641 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     2701 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2761 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2821 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2881 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2941 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3001 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3061 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3121 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3181 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3241 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3301 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3361 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3421 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3481 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3541 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     3601 cgcgcgttgg ccgattcatt aatgcag
//
