LOCUS       sequence-180921-        3460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of a fluorescent protein, monomeric Honeydew 
            (mHoneydew).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3460)
  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 3460)
  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 mHoneydew
FEATURES             Location/Qualifiers
     source          1..3460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(33..50)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(204..792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(284..303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(963..1754,1755..1823))
                     /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     1586..1605
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1824..1928)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1954..2409)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2091..2112)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2303..2322
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(2506..2553)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2549..2567
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(2633..3328)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mHoneydew
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMWGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWAATTERMYPEDGALKGEIK
                     MRLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKIDGKLDITSHNEDYTIVEQYERAEGRH
                     STGGMDELYK"
     primer_bind     complement(2710..2733)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2908..2927)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3177..3195
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     RBS             3336..3358
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3389..3408)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3390..3408)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
       61 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
      121 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      181 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      241 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      301 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      361 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
      421 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
      481 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
      541 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
      601 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
      661 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
      721 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
      781 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
      841 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
      901 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
      961 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     1021 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     1081 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     1141 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     1201 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     1261 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     1321 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     1381 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     1441 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     1501 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     1561 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     1621 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     1681 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     1741 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     1801 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     1861 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     1921 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     1981 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     2041 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     2101 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     2161 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     2221 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     2281 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     2341 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     2401 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     2461 cctcttcgct attacgccag atccggatat agttcctcct ttcagcaaaa aacccctcaa
     2521 gacccgttta gaggccccaa ggggttatgc tagttattgc tcagcggtgg cagcagccaa
     2581 ctcagcttcc tttcgggctt tgttagcagc cggatcaagc ttcgaattct tacttgtaca
     2641 gctcgtccat gccgccggtg gagtggcggc cctcggcgcg ctcgtactgt tccacgatgg
     2701 tgtagtcctc gttgtgggag gtgatgtcca gcttcccgtc aatcttgtag gcgccgggca
     2761 gctgcacggg cttcttggcc atgtaggtgg tcttgacctc ggcgtcgtag tggccgccgt
     2821 ccttcagctt cagcctcatc ttgatctcgc ccttcagggc gccgtcctcg gggtacatcc
     2881 gctcggtggt ggccgcccag cccatggtct tcttctgcat tacggggccg tcggagggga
     2941 agttggtgcc gcgcagcttc accttgtaga tgaactcgcc gtcctgcagg gaggagtcct
     3001 gggtcacggt caccacgccg ccgtcctcga agttcatcac gcgctcccac ttgaagccct
     3061 cggggaagga cagcttcaag tagtcgggga tgtcggcggg gtgcttcacg taggccttgg
     3121 agccccacat gaactgaggg gacaggatgt cccaggcgaa gggcaggggg ccgcccttgg
     3181 tcaccttcag cttggcggtc tgggtgccct cgtaggggcg gccctcgccc tcgccctcga
     3241 tctcgaactc gtggccgttc acggagccct ccatgcgcac cttgaagcgc atgaactcct
     3301 tgatgacctc ctcgcccttg ctcaccatat gtatatctcc ttcttaaagt taaacaaaat
     3361 tatttctaga gggaaaccgt tgtggtctcc ctatagtgag tcgtattaat ttcgcgggat
     3421 cgagatcctg cattaatgaa tcggccaacg cgcggggaga
//
