LOCUS       sequence-181593-        3541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of the primed conversion capable protein 
            pr-Kaede (Kaede-A69T)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3541)
  AUTHORS   Mohr MA, Kobitski AY, Sabater LR, Nienhaus K, Obara CJ, 
            Lippincott-Schwartz J, Nienhaus GU, Pantazis P
  TITLE     Rational Engineering of Photoconvertible Fluorescent Proteins for 
            Dual-Color Fluorescence Nanoscopy Enabled by a Triplet-State 
            Mechanism of Primed Conversion.
  JOURNAL   Angew Chem Int Ed Engl. 2017 Jun 29. doi: 10.1002/anie.201706121.
  PUBMED    28661566
REFERENCE   2  (bases 1 to 3541)
  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: "Angew Chem 
            Int Ed Engl. 2017 Jun 29. doi: 10.1002/anie.201706121."
COMMENT     Sequence Label: pRSET-pr-Kaede (Kaede-A69T)
FEATURES             Location/Qualifiers
     source          1..3541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     256..275
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        256..274
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             306..328
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             348..365
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             369..401
                     /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(405..413,414..428)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             435..1115
                     /codon_start=1
                     /product="Kaede green-to-red photoconvertible fluorescent 
                     protein (Ando et al., 2002)"
                     /label=Kaede
                     /translation="MVSLIKPEMKIKLLMEGNVNGHQFVIEGDGKGHPFEGKQSMDLVV
                     KEGAPLPFAYDILTTAFHYGNRVFTKYPDHIPDYFKQSFPKGFSWERSLMFEDGGVCIA
                     TNDITLKGDTFFNKVRFDGVNFPPNGPVMQKKTLKWEASTEKMYLRDGVLTGDITMALL
                     LKGDVHYRCDFRTTYKSRQEGVKLPGYHFVDHCISILRHDKDYNEVKLYEHAVAHSGLP
                     DNVK"
     primer_bind     complement(1178..1196)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1192..1239
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1336..1791
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1423..1442)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1633..1654
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1817..1921
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1922..1990,1991..2782)
                     /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(2140..2159)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2953..3541
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3442..3461
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcaggatc
      241 tcgatcccgc gaaattaata cgactcacta tagggagacc acaacggttt ccctctagaa
      301 ataattttgt ttaactttaa gaaggagata tacatatgcg gggttctcat catcatcatc
      361 atcatggtat ggctagcatg actggtggac agcaaatggg tcgggatctg tacgacgatg
      421 acgataagga tccgatggtg agtctgatta aaccagaaat gaagatcaag ctgcttatgg
      481 aaggcaatgt aaacgggcac cagtttgtta ttgagggaga tggaaaaggc catccttttg
      541 agggaaaaca gagtatggac cttgtagtca aagaaggcgc acctctccct tttgcctacg
      601 atatcttgac aacagcattc cattatggta acagggtttt taccaaatac ccagaccata
      661 taccagacta cttcaagcag tcgtttccca aagggttttc ttgggagcga agcctgatgt
      721 tcgaggacgg gggcgtttgc atcgctacaa atgacataac actgaaagga gacacttttt
      781 ttaacaaagt tcgatttgat ggcgtaaact ttcccccaaa tggtcctgtt atgcagaaga
      841 agactctgaa atgggaggca tccactgaga aaatgtattt gcgtgatgga gtgttgacgg
      901 gcgatattac aatggctctg ctgcttaaag gagatgtcca ttaccgatgt gacttcagaa
      961 ctacttacaa atctaggcag gagggtgtca agttgccagg atatcacttt gtcgatcact
     1021 gcatcagcat attgaggcat gacaaagact acaacgaggt taagctgtat gagcacgctg
     1081 ttgcccattc tggattgccg gacaacgtca agtaagaatt cgaagcttga tccggctgct
     1141 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     1201 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     1261 ggatctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     1321 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     1381 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     1441 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     1501 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     1561 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     1621 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     1681 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     1741 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     1801 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     1861 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     1921 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     1981 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2041 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     2101 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2161 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2221 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2281 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2341 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2401 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     2461 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     2521 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     2581 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     2641 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     2701 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     2761 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     2821 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     2881 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2941 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3001 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3061 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3121 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3181 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3241 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3301 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3361 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3421 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3481 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3541 a
//
