LOCUS       sequence-288032-        3580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of Sirius, a pH-insensitive and photostable 
            ultramarine fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3580)
  AUTHORS   Tomosugi W, Matsuda T, Tani T, Nemoto T, Kotera I, Saito K, Horikawa
            K, Nagai T
  TITLE     An ultramarine fluorescent protein with increased photostability and
            pH insensitivity.
  JOURNAL   Nat Methods. 2009 May;6(5):351-3. doi: 10.1038/nmeth.1317. Epub 2009
            Apr 6.
  PUBMED    19349978
REFERENCE   2  (bases 1 to 3580)
  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: 
            "10.1038/nmeth.1317"; journalName: "Nat Methods"; date: "2009-05"; 
            volume: "6"; issue: "5"; pages: "351-3"
COMMENT     Sequence Label: Sirius/pRSETB
FEATURES             Location/Qualifiers
     source          1..3580
                     /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             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             112..129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             133..165
                     /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(169..177,178..192)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(244..265)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(505..524)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             664..915
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKAHFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSVQSKLSKDPNEKRDHMVLLESVTAAGITLGMDELYK"
     primer_bind     complement(981..999)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      995..1042
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1139..1594
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1226..1245)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1436..1457
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1620..1724
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1725..1793,1794..2585)
                     /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(1943..1962)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2756..3344
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3245..3264
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3498..3515
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
      181 gatgacgata aggatccgat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      241 atcctggtcg agctggacgg cgacgtaaac ggccacaggt tcagcgtgtc cggcgagggc
      301 gagggcgatg ccacctacgg caagctgacc ctgaagctca tctgcaccac cggcaagctg
      361 cccgtgccct ggcccaccct cgtgaccacc ctgcaattcg gcgtgctgtg cttcgcccgc
      421 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      481 caggagcgta ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      541 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      601 ggcaacatcc tggggcacaa gctggagtac aacgggataa gctcaaacgt atatatcacc
      661 gccgacaagc agaagaacgg catcaaggcc cacttcaaga tccgccacaa catcgaggac
      721 ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
      781 ctgctgcccg acaaccacta cctgagcgtc cagtccaagc tgagcaaaga ccccaacgag
      841 aagcgcgatc acatggtcct gctggagtcc gtgaccgccg ccgggatcac tctcggcatg
      901 gacgagctgt acaagtaaga attcgaagct tgatccggct gctaacaaag cccgaaagga
      961 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     1021 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatctg gcgtaatagc
     1081 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     1141 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1201 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1261 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1321 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1381 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1441 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1501 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1561 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     1621 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1681 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1741 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1801 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     1861 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     1921 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     1981 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2041 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2101 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2161 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2221 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2281 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2341 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2401 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2461 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2521 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2581 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2641 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     2701 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2761 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2821 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2881 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2941 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3001 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3061 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3121 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3181 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3241 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3301 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3361 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3421 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3481 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     3541 aaccgcctct ccccgcgcgt tggccgattc attaatgcag
//
