LOCUS       sequence-180849-        3577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of a fluorescent protein, monomeric Strawberry 
            (mStrawberry).
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 mStrawberry
FEATURES             Location/Qualifiers
     source          1..3577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     28..45
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     130..149
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        130..148
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             180..202
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             222..239
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             243..275
                     /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(279..287,288..302)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      309..318
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             315..1025
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent 
                     protein"
                     /label=mStrawberry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILTPNFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKMRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVELYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(460..478)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     728..747
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1088..1106)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1102..1149
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1246..1701
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1333..1352)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1543..1564
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1727..1831
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1832..1900,1901..2692)
                     /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(2050..2069)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2863..3451
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3352..3371
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
       61 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag gatctcgatc
      121 ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct agaaataatt
      181 ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat catcatcatg
      241 gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac gatgacgata
      301 aggatcccgc caccatggtg agcaagggcg aggagaataa catggccatc atcaaggagt
      361 tcatgcgctt caaggtgcgc atggagggct ccgtgaacgg ccacgagttc gagatcgagg
      421 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
      481 gtggccccct gcccttcgcc tgggacatcc taacccccaa cttcacctac ggctccaagg
      541 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
      601 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
      661 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
      721 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
      781 accccgagga cggcgccctg aagggcgaga tcaagatgag gctgaagctg aaggacggcg
      841 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
      901 gcgcctacat cgtcggcatc aagttggaca tcacctccca caacgaggac tacaccatcg
      961 tggaactgta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     1021 agtaagaatt cgaagcttga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     1081 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     1141 ggttttttgc tgaaaggagg aactatatcc ggatctggcg taatagcgaa gaggcccgca
     1201 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     1261 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1321 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     1381 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     1441 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     1501 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     1561 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     1621 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     1681 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1741 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     1801 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     1861 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     1921 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     1981 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     2041 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     2101 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2161 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2221 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2281 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2341 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     2401 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     2461 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2521 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2581 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2641 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     2701 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     2761 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2821 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2881 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2941 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3001 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3061 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3121 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3181 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3241 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3301 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3361 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3421 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3481 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3541 ggataaccgt attaccgcct ttgagtgagc tgatacc
//
