LOCUS       sequence-180920-        3577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of a fluorescent protein, monomeric Orange 
            (mOrange).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3577)
  AUTHORS   Shaner NC, Campbell RE, Steinbach PA, Giepmans BN, Palmer AE, Tsien 
            RY
  TITLE     Improved monomeric red, orange and yellow fluorescent proteins 
            derived from Discosoma sp. red fluorescent protein.
  JOURNAL   Nat Biotechnol. 2004 Dec;22(12):1567-72. Epub 2004 Nov 21.
  PUBMED    15558047
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"; journalName: "Nat 
            Biotechnol."; date: "2004-12"; volume: "22(12)"; pages: "1567-72. 
            Epub 2004 Nov 21"
COMMENT     Sequence Label: pNCS mOrange
FEATURES             Location/Qualifiers
     source          1..3577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             31..53
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             73..90
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             94..126
                     /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(130..138,139..153)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      160..169
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             166..876
                     /codon_start=1
                     /product="orange monomeric derivative of DsRed fluorescent 
                     protein (Shanet et al., 2004)"
                     /label=mOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     579..598
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(939..957)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      953..1000
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1097..1552
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1184..1203)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1394..1415
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1578..1682
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1683..1751,1752..2543)
                     /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(1901..1920)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2714..3302
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3203..3222
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3456..3473
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3558..3577
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3558..3576
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 agaccacaac ggtttccctc tagaaataat tttgtttaac tttaagaagg agatatacat
       61 atgcggggtt ctcatcatca tcatcatcat ggtatggcta gcatgactgg tggacagcaa
      121 atgggtcggg atctgtacga cgatgacgat aaggatcccg ccaccatggt gagcaagggc
      181 gaggagaata acatggccat catcaaggag ttcatgcgct tcaaggtgcg catggagggc
      241 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
      301 tttcagaccg ctaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
      361 ctgtcccctc agttcaccta cggctccaag gcctacgtga agcaccccgc cgacatcccc
      421 gactacttca agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
      481 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
      541 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
      601 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
      661 atcaagatga ggctgaagct gaaggacggc ggccactaca cctccgaggt caagaccacc
      721 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca tcgtcggcat caagttggac
      781 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
      841 cactccaccg gcggcatgga cgagctgtac aagtaagaat tcgaagcttg atccggctgc
      901 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
      961 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     1021 cggatctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     1081 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     1141 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     1201 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     1261 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     1321 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     1381 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     1441 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     1501 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     1561 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1621 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1681 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1741 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1801 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     1861 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     1921 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     1981 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2041 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2101 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2161 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     2221 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     2281 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     2341 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2401 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2461 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2521 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     2581 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2641 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2701 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2761 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2821 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2881 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2941 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3001 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3061 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3121 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3181 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3241 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3301 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3361 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3421 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3481 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagga
     3541 tctcgatccc gcgaaattaa tacgactcac tataggg
//
