LOCUS       sequence-181572-        4133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of the primed conversion capable protein 
            pr-mEosFP (mEosFP-V69T)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4133)
  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 4133)
  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: pQE32-pr-mEosFP (mEosFP-V69T)
FEATURES             Location/Qualifiers
     source          1..4133
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..24,25..30,31..47,48..54)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    62..78
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             127..144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             154..834
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="MSAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFTEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFHGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIHMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     terminator      880..974
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1018..1677
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1084..1103)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1742..1828
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1875..1897)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1983..2123
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2038..2057
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2138..2155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2309..2897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2389..2408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3068..3859,3860..3928))
                     /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     3691..3710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3929..4033)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4101..4119
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
      121 ggatctcacc atcaccatca ccatgggatc catatgagtg cgattaagcc agacatgaag
      181 atcaacctcc gtatggaagg caacgtaaac gggcaccact ttgtgatcga cggagatggt
      241 acaggcaagc cttttgaggg aaaacagagt atggatcttg aagtcaaaga gggcggacct
      301 ctgccttttg cctttgatat cctgaccact gcattccatt acggcaacag ggtattcacc
      361 gaatatccag accacataca agactatttt aagcagtcgt ttcctaaggg gtattcgtgg
      421 gaacgaagct tgactttcga agacgggggc atttgcattg ccagaaacga cataacaatg
      481 gaaggggaca ctttctataa taaagttcga tttcacggta caaactttcc cgccaatggt
      541 ccagttatgc agaagaagac gctgaaatgg gagccctcca ctgagaaaat gtatgtgcgt
      601 gatggagtgc tgacgggtga tattcacatg gctttgttgc ttgaaggaaa tgcccattac
      661 cgatgtgact tcagaactac ttacaaagct aaggagaagg gtgtcaagtt accaggctac
      721 cactttgtgg accactgcat tgagatttta agccatgaca aagattacaa caaggttaag
      781 ctgtatgagc atgctgttgc tcattctgga ttgcctgaca atgccagacg ataaggtacc
      841 ccgggtcgac ctgcagccaa gcttaattag ctgagcttgg actcctgttg atagatccag
      901 taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt
      961 ttattggtga gaatccaagc tagcttggcg agattttcag gagctaagga agctaaaatg
     1021 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
     1081 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
     1141 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
     1201 attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag
     1261 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
     1321 ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
     1381 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
     1441 atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
     1501 aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac
     1561 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
     1621 ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt
     1681 ttttaaggca gttattggtg cccttaaacg cctggggtaa tgactctcta gcttgaggca
     1741 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     1801 ggtgaacgct ctcctgagta ggacaaatcc gccctctaga gctgcctcgc gcgtttcggt
     1861 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     1921 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     1981 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
     2041 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     2101 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2161 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2221 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2281 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2341 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2401 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2461 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2521 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2581 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2641 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2701 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     2761 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2821 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2881 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2941 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3001 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3061 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3121 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3181 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3241 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3301 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3361 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3421 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3481 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3541 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3601 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3661 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3721 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3781 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3841 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3901 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3961 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4021 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     4081 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt cac
//
