LOCUS       sequence-474128-        3583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of rsZACRO, a positively reversibly 
            photoswitchable fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3583)
  AUTHORS   Ozaki-Noma R, Wazawa T, Kakizuka T, Shidara H, Takemoto K, Nagai T
  TITLE     Positive-Type Reversibly Photoswitching Red Fluorescent Protein for 
            Dual-Color Superresolution Imaging with Single Light Exposure for 
            Off-Switching.
  JOURNAL   ACS Nano. 2025 Feb 25;19(7):7188-7201. doi: 10.1021/acsnano.4c16847.
            Epub 2025 Feb 12.
  PUBMED    39937184
REFERENCE   2  (bases 1 to 3583)
  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/acsnano.4c16847"; journalName: "ACS Nano"; date: 
            "2025-02-25- 25"; volume: "19"; issue: "7"; pages: "7188-7201"
COMMENT     Sequence Label: rsZACRO/pRSETB
FEATURES             Location/Qualifiers
     source          1..3583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             112..129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             133..165
                     /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(169..177,178..192)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(984..1002)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      998..1045
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1142..1597
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1229..1248)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1439..1460
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1623..1727
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1728..1796,1797..2588)
                     /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(1946..1965)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2759..3347
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3248..3267
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3501..3518
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
      181 gatgacgata aggatcccac catgggttcg gaggtcggcc ccgccctgtt ccagcgggac
      241 atgaccttca aaatcttcat cgacggcgag gttaacggcc agaagttcac catcgtggcc
      301 gacggcagca gcaagttccc ccacggcgac ttcaacgtgc acgccgtgtg cgagaccggc
      361 aagctgccca tgagctggaa gcccatctgc cacctgatcc agtacggcga gcccttcttc
      421 gcccgctacc ccgacggcat catccatttc agtcaggagt gcttccccga gggcctgagc
      481 atcgaccgca ccgtgcgctt cgagaacgac ggcaccatga ccagccacca cacctacgag
      541 ctggacggca cctgcgtggt gagccgcatc accgtgaact gcgacggctt cctgcccgac
      601 ggccccatca tgcgcgacca gctggtggac attcgtccct gcgagacccg catactcccc
      661 tacggcccca acgccgtgcg ccagaccgcc accatgggct tcaccaccgc cgacggcggc
      721 aaggtgatgg gccacttcga cagcaagatg accttcaacg gcagccgcgc catcgagatc
      781 cccggcccac acttcgtgac catcatcacc gagcagacga gggaccctag cgacaagcgc
      841 gaccacgtgt gccagcgcga ggtggcctac gcccacagcg ggccccgcaa caccagcgcc
      901 atcggtagcg acgaggatta agaattcgaa gcttgatccg gctgctaaca aagcccgaaa
      961 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     1021 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat ctggcgtaat
     1081 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     1141 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     1201 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     1261 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     1321 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     1381 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     1441 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     1501 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     1561 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     1621 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1681 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1741 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1801 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1861 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     1921 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     1981 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     2041 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     2101 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     2161 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2221 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     2281 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     2341 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     2401 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     2461 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     2521 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     2581 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     2641 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     2701 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2761 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2821 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2881 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2941 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3001 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3061 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3121 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3181 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3241 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3301 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3361 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3421 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3481 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     3541 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cag
//
