LOCUS       sequence-354479-        3630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of mClover3 in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3630)
  AUTHORS   Bajar BT, Wang ES, Lam AJ, Kim BB, Jacobs CL, Howe ES, Davidson MW, 
            Lin MZ, Chu J
  TITLE     Improving brightness and photostability of green and red fluorescent
            proteins for live cell imaging and FRET reporting.
  JOURNAL   Sci Rep. 2016 Feb 16;6:20889. doi: 10.1038/srep20889.
  PUBMED    26879144
REFERENCE   2  (bases 1 to 3630)
  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: "Sci Rep."; 
            date: "2016-02-16"; volume: "6:20889. doi"; pages: " 
            10.1038/srep20889"
COMMENT     Sequence Label: pNCS-mClover3
FEATURES             Location/Qualifiers
     source          1..3630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(154..742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(234..253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(913..1704,1705..1773))
                     /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     1536..1555
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1774..1878)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1904..2359)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2041..2062)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2253..2272
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(2456..2503)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2499..2517
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(join(2578..3291,3292..3294,3295..3297))
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2623..2644)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3231..3252
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(join(3304..3318,3319..3327))
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             complement(3331..3363)
                     /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             complement(3367..3384)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3509..3528)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3510..3528)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3613..3630)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
       61 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
      121 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
      181 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
      241 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      301 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      361 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      421 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
      481 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
      541 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
      601 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
      661 atccggcaaa aaaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
      721 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
      781 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
      841 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
      901 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
      961 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     1021 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     1081 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     1141 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     1201 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     1261 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     1321 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     1381 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     1441 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     1501 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     1561 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     1621 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     1681 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     1741 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     1801 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     1861 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     1921 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     1981 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2041 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2101 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     2161 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2221 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2281 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2341 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     2401 tcggtgcggg cctcttcgct attacgccag atccggatat agttcctcct ttcagcaaaa
     2461 aacccctcaa gacccgttta gaggccccaa ggggttatgc tagttattgc tcagcggtgg
     2521 cagcagccaa ctcagcttcc tttcgggctt tgttagcagc cggatcaagc ttcgaattca
     2581 cttgtacagc tcgtccatgc catgtgtaat cccggcggcg gtcacgaact ccagcaggac
     2641 catgtgatcg cgcttctcgt tggggtcttt gctcagcttg gactgatggc tcaggtagtg
     2701 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
     2761 gagctgcacg ctgccgtcct caacgttgtg gcggatcttg aagttagcct tgatgcagtt
     2821 cttctgcttg tcggccgtga tatagacgta gtggctgttg aagttgtact ccagcttgtg
     2881 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
     2941 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag gtaccgtcgt ccttgaaaga
     3001 gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt
     3061 catgtggtcg gggtagcggc tgaagcaggc cacgccgtag ccgaaggtgg tcacgagggt
     3121 gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgtt
     3181 ggtggcatcg ccctcgccct cgccgcggac gctgaacttg tggccgttta cgtcgccgtc
     3241 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatcgg
     3301 atccttatcg tcatcgtcgt acagatcccg acccatttgc tgtccaccag tcatgctagc
     3361 cataccatga tgatgatgat gatgagaacc ccgcatatgt atatcccctt cttaaagtta
     3421 aacaaaacag tactttccta ttatatcatt catcgcatca ggtgtcaaga ataaactcgg
     3481 tgattctaga gggaaaccgt tgtggtctcc ctatagtgag tcgtattaat ttcgcgggat
     3541 cgagatcctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3601 ctcttccgct tcctcgctca ctgactcgct
//
