LOCUS       sequence-476344-        3550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of RSFP pcStar; photoconvertible red 
            fluorescent protein with high brightness, high photoconversion 
            contrast, and early detectable fluorescence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3550)
  AUTHORS   Zhang M, Fu Z, Li C, Liu A, Peng D, Xue F, He W, Gao S, Xu F, Xu D, 
            Yuan L, Zhang F, Xu Z, Xu T, Xu P
  TITLE     Fast Super-Resolution Imaging Technique and Immediate Early 
            Nanostructure Capturing by a Photoconvertible Fluorescent Protein.
  JOURNAL   Nano Lett. 2020 Apr 8;20(4):2197-2208. doi: 
            10.1021/acs.nanolett.9b02855. Epub 2019 Oct 7.
  PUBMED    31576756
REFERENCE   2  (bases 1 to 3550)
  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"; doi: 
            "10.1021/acs.nanolett.9b02855"; journalName: "Nano Lett"; date: 
            "2020-04-8- 8"; volume: "20"; issue: "4"; pages: "2197-2208"
COMMENT     Sequence Label: pRSET A-pcStar
FEATURES             Location/Qualifiers
     source          1..3550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(684..698,699..707))
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             complement(711..743)
                     /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(747..764)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             784..806
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(837..856)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(838..856)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(941..958)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1112..1700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1192..1211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1871..2662,2663..2731))
                     /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     2494..2513
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2732..2836)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2862..3317)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2999..3020)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     terminator      complement(3414..3461)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3457..3475
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(join(3538..3550,1..668))
                     /codon_start=1
                     /product="improved monomeric variant of green-to-red 
                     photoswitchable fluorescent protein EosFP (Zhang et al., 
                     2012)"
                     /label=mEos3.2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGEGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSMTFEDGGICNAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALLL
                     EGGAHYRCDFRTTYKAKEKGVKLPGAHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
ORIGIN
        1 ttgtcaggca atccagaatg agcaacagca tgctcataca gcttaacctt gttgtaatct
       61 ttgtcatggc ttaaaatctc aatgcagtgg tccacaaagt gggcgcctgg taacttgaca
      121 cccttctcct tagctttgta agtagttctg aagtcacatc ggtaatgggc acctccttca
      181 agcaacaaag ccatctcaat atcacccgtc agcactccat cacgcacata cattttctca
      241 gtggagggct cccatttcag cgtcttcttc tgcataactg gaccattggc gggaaagttg
      301 gtaccataaa atcgaacttt attatagaaa gtgtcccctt ccattgttat gtcgtttctg
      361 gcgttgcaaa tgcccccgtc ttcgaaagtc atgcttcgtt cccacgaata ccccttagga
      421 aacgactgct taaaatagtc ttgtatgttg tctggatatt tggcgaatac cctgttgccg
      481 taatggaatg cagtggtcag gatatcaaag gcaaaaggca gaggtccgcc ctctttgact
      541 tcaagatcca tactctgttt tccctcaaaa ggcttgcctg taccttctcc gtcgatcaca
      601 aagtggtgcc cgtttacgtt gccttccata cggagtttga tcttcatgtc tggcttaatc
      661 gcactcatgg atccccatcg atccttatcg tcatcgtcgt acagatcccg acccatttgc
      721 tgtccaccag tcatgctagc cataccatga tgatgatgat gatgagaacc ccgcatatgt
      781 atatctcctt cttaaagtta aacaaaatta tttctagagg gaaaccgttg tggtctccct
      841 atagtgagtc gtattaattt cgcgggatcg agatcctgca ttaatgaatc ggccaacgcg
      901 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
      961 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1021 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1081 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1141 atcacaaaaa tcgacgctca agtcagaggt gtcgaaaccc gacaggacta taaagatacc
     1201 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1261 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1321 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1381 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1441 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1501 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     1561 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1621 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1681 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     1741 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     1801 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     1861 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     1921 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     1981 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     2041 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     2101 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     2161 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     2221 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     2281 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     2341 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     2401 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     2461 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     2521 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     2581 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     2641 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     2701 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     2761 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     2821 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     2881 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2941 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3001 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3061 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3121 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3181 gggaaagccg gcgaacgtgg cgagaaagga agggaataaa gcgaaaggag cgggcgctag
     3241 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3301 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     3361 ggtgcgggcc tcttcgctat tacgccagat ccggatatag ttcctccttt cagcaaaaaa
     3421 cccctcaaga cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca
     3481 gcagccaact cagcttcctt tcgggctttg ttagcagccg gatcaagctt cgaattctta
     3541 tcgtctggca
//
