LOCUS       sequence-176636-        4123 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Orange color variant of bright luminescent protein for bacterial 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4123)
  AUTHORS   Suzuki K, Kimura T, Shinoda H, Bai G, Daniels MJ, Arai Y, Nakano M, 
            Nagai T
  TITLE     Five colour variants of bright luminescent protein for real-time 
            multicolour bioimaging.
  JOURNAL   Nat Commun. 2016 Dec 14;7:13718. doi: 10.1038/ncomms13718.
  PUBMED    27966527
REFERENCE   2  (bases 1 to 4123)
  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 
            Commun."; date: "2016-12-14"; volume: "7:13718. doi"; pages: " 
            10.1038/ncomms13718"
COMMENT     Sequence Label: OeNL_pRSETb
FEATURES             Location/Qualifiers
     source          1..4123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1055..1073)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1069..1116
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1213..1668
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1300..1319)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1510..1531
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1694..1798
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1799..1867,1868..2659)
                     /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(2017..2036)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2830..3418
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3319..3338
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3572..3589
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3674..3693
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3674..3692
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3724..3746
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3766..3783
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3787..3819
                     /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(3823..3831,3832..3846)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(4093..4123,1..620)
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="VSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRVT
                     MAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSAS
                     VSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMYL
                     KLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     CDS             join(4093..4123,1..620)
                     /codon_start=1
                     /product="humanized monomeric Kusabira-Orange fluorescent 
                     protein"
                     /label=hmKO
                     /note="mammalian codon-optimized"
                     /translation="VSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRVT
                     MAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSAS
                     VSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMYL
                     KLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
ORIGIN
        1 ggtactacat ggacggctcc gtcaatgggc atgagttcac aatcgagggt gagggcacag
       61 gcagacctta cgagggacat caggagatga cactgcgcgt cacaatggcc gagggcgggc
      121 caatgccttt cgccttcgac ctggtgtccc acgtgttctg ttacggccac agagttttta
      181 ctaaatatcc agaagagatc ccagactatt tcaagcaggc ctttcctgag ggcctgtcct
      241 gggagaggtc cctggagttc gaggacggcg gctccgcctc cgtgagcgcc cacatcagcc
      301 tgaggggcaa caccttctac cacaagtcca agttcaccgg cgtgaacttc cccgccgacg
      361 gccccatcat gcagaaccag agcgtggact gggagccctc caccgagaag atcaccgcca
      421 gcgacggcgt gctgaagggc gacgtgacca tgtacctgaa gctggagggc ggcggcaacc
      481 acaagtgcca gatgaagacc acctacaagg ccgccaagga gatcctggag atgcccggcg
      541 accactacat cggccacagg ctggtgagga agaccgaggg caacatcacc gagcaggtgg
      601 aggacgccgt ggcccactcc gagctcgaaa atgggctgaa gatcgacatc catgtcatca
      661 tcccgtatga aggtctgagc ggcgaccaaa tgggccagat cgaaaaaatt tttaaggtgg
      721 tgtaccctgt ggatgatcat cactttaagg tgatcctgca ctatggcaca ctggtaatcg
      781 acggggttac gccgaacatg atcgactatt tcggacggcc gtatgaaggc atcgccgtgt
      841 tcgacggcaa aaagatcact gtaacaggga ccctgtggaa cggcaacaaa attatcgacg
      901 agcgcctgat caaccccgac ggctccctgc tgttccgagt aaccatcaac ggagtgaccg
      961 gctggcggct gtgcgaacgc attctggcgt aagaattcga agcttgatcc ggctgctaac
     1021 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     1081 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     1141 tctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     1201 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1261 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     1321 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     1381 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     1441 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     1501 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     1561 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     1621 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     1681 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1741 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     1801 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     1861 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     1921 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     1981 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     2041 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     2101 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     2161 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     2221 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     2281 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     2341 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     2401 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     2461 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     2521 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2581 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     2641 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     2701 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     2761 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2821 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2881 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2941 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3001 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3061 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3121 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3181 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3241 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3301 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3361 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3421 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     3481 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     3541 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     3601 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcaggatctc
     3661 gatcccgcga aattaatacg actcactata gggagaccac aacggtttcc ctctagaaat
     3721 aattttgttt aactttaaga aggagatata catatgcggg gttctcatca tcatcatcat
     3781 catggtatgg ctagcatgac tggtggacag caaatgggtc gggatctgta cgacgatgac
     3841 gataaggatc cgatggtctt cacactcgaa gatttcgttg gggactggcg acagacagcc
     3901 ggctacaacc tggaccaagt ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc
     3961 ggggtgtccg taactccgat ccaaaggatt gtcctgagcg gtctcgaggg cggtagcggt
     4021 ggcagcgtga gcgtgatcaa gcccgagatg aagaaggtgg aggacgccgt ggcccactcc
     4081 actctcgagg gcgtgagcgt gatcaagccc gagatgaaga tga
//
