LOCUS       sequence-77348-e        3606 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Photoconverible Fluorescent Protein_ Fixative Resistant (monomeric) 
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3606)
  AUTHORS   Paez-Segala MG, Sun MG, Shtengel G, Viswanathan S, Baird MA, Macklin
            JJ, Patel R, Allen JR, Howe ES, Piszczek G, Hess HF, Davidson MW, 
            Wang Y, Looger LL
  TITLE     Fixation-resistant photoactivatable fluorescent proteins for CLEM.
  JOURNAL   Nat Methods. 2015 Jan 12. doi: 10.1038/nmeth.3225.
  PUBMED    25581799
REFERENCE   2  (bases 1 to 3606)
  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 
            Methods. 2015 Jan 12. doi: 10.1038/nmeth.3225."
COMMENT     Sequence Label: pRSETa_ mEos4b
FEATURES             Location/Qualifiers
     source          1..3606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..38
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        19..37
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             68..90
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             110..127
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             131..163
                     /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(167..175,176..190)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             206..889
                     /codon_start=1
                     /product="fixation-resistant truly monomeric green-to-red 
                     photoswitchable fluorescent protein (Paez-Segala et al., 
                     2015)"
                     /label=mEos4b
                     /note="derived from mEos2"
                     /translation="MVSAIKPDMRIKLRMEGNVNGHHFVIDGDGTGKPYEGKQTMDLEV
                     KEGGPLPFAFDILTTAFHYGNRVFVKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNA
                     RNDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIEMALL
                     LEGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHAIEILSHDKDYNKVKLYEHAVAHSGLP
                     DNARR"
     primer_bind     complement(952..970)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      966..1013
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1110..1565
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1197..1216)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1407..1428
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1646..1750
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1751..1819,1820..2611)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRPPRYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLID*MEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2782..3370
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3271..3290
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3524..3541
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
       61 gaataatttt gtttaacttt aagaaggaga tatacatatg cggggttctc atcatcatca
      121 tcatcatggt atggctagca tgactggtgg acagcaaatg ggtcgggatc tgtacgacga
      181 tgacgataag gatcgatggg gatccatggt gagtgcgatt aagccagaca tgaggatcaa
      241 actccgtatg gaaggcaacg taaacgggca ccactttgtg atcgacggag atggtacagg
      301 caagccttat gagggaaaac agaccatgga tcttgaagtc aaagagggcg gacctctgcc
      361 ttttgccttt gatatcctga ccactgcatt ccattacggc aacagggtat tcgtgaaata
      421 tccagacaac atacaagact attttaagca gtcgtttcct aaggggtatt cgtgggaacg
      481 aagcttgact ttcgaagacg ggggcatttg caatgccaga aacgacataa caatggaagg
      541 ggacactttc tataataaag ttcgatttta tggtaccaac tttcccgcca atggtccagt
      601 tatgcagaag aagacgctga aatgggagcc ctccactgag aaaatgtatg tgcgtgatgg
      661 agtgctgacg ggtgatattg agatggcttt gttgcttgaa ggaaatgccc attaccgatg
      721 tgacttcaga actacttaca aagctaagga gaagggtgtc aagttaccag gcgcccactt
      781 tgtggaccac gccattgaga ttttaagcca tgacaaagat tacaacaagg ttaagctgta
      841 tgagcatgct gttgctcatt ctggattgcc tgacaatgcc agacgataag aattcgaagc
      901 ttgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
      961 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     1021 gaggaactat atccggatct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     1081 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     1141 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     1201 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     1261 tcgggggctc cctttagggt tccgatttag agctttacgg cacctcgacc gcaaaaaact
     1321 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     1381 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     1441 ccctatcgcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     1501 aaaaaatgag ctgatttaac aaatatttaa cgcgaatttt aacaaaatat taacgtttac
     1561 aatttcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     1621 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1681 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1741 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1801 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1861 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1921 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtgatac
     1981 actattatcc cgtattgacg ccgggcaaga gcaactcggt cgccccccgc ggtattctca
     2041 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2101 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2161 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2221 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagagtga
     2281 caccacgatg cctgtagcaa tgccaacaac gttgcgcaaa ctattaactg gcgaactact
     2341 tactctagct tcccggcaac aattaataga ctgaatggag gcggataaag ttgcaggacc
     2401 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2461 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagcgct cccgtatcgt
     2521 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2581 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2641 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2701 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2761 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2821 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2881 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     2941 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3001 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3061 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3121 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3181 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3241 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3301 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3361 cctatggaaa aacgccagca acgcggcctt tttacggttc ctgggctttt gctggccttt
     3421 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3481 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3541 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     3601 atgcag
//
