LOCUS       sequence-213218-        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(1..15,16..21,22..38,39..45)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    21..37
                     /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    53..69
                     /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             88..99
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             118..135
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             145..825
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="MSAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFTEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFHGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIHMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     terminator      871..965
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1009..1668
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1075..1094)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1733..1819
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1866..1888)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1974..2114
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2029..2048
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2129..2146)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2300..2888)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2380..2399)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3059..3850,3851..3919))
                     /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     3682..3701
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3920..4024)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4092..4110
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc aattgtgagc
       61 ggataacaat ttcacacaga attcattaaa gaggagaaat taactatgag aggatctcac
      121 catcaccatc accatgggat ccatatgagt gcgattaagc cagacatgaa gatcaacctc
      181 cgtatggaag gcaacgtaaa cgggcaccac tttgtgatcg acggagatgg tacaggcaag
      241 ccttttgagg gaaaacagag tatggatctt gaagtcaaag agggcggacc tctgcctttt
      301 gcctttgata tcctgaccac tgcattccat tacggcaaca gggtattcac cgaatatcca
      361 gaccacatac aagactattt taagcagtcg tttcctaagg ggtattcgtg ggaacgaagc
      421 ttgactttcg aagacggggg catttgcatt gccagaaacg acataacaat ggaaggggac
      481 actttctata ataaagttcg atttcacggt acaaactttc ccgccaatgg tccagttatg
      541 cagaagaaga cgctgaaatg ggagccctcc actgagaaaa tgtatgtgcg tgatggagtg
      601 ctgacgggtg atattcacat ggctttgttg cttgaaggaa atgcccatta ccgatgtgac
      661 ttcagaacta cttacaaagc taaggagaag ggtgtcaagt taccaggcta ccactttgtg
      721 gaccactgca ttgagatttt aagccatgac aaagattaca acaaggttaa gctgtatgag
      781 catgctgttg ctcattctgg attgcctgac aatgccagac gataaggtac cccgggtcga
      841 cctgcagcca agcttaatta gctgagcttg gactcctgtt gatagatcca gtaatgacct
      901 cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg
      961 agaatccaag ctagcttggc gagattttca ggagctaagg aagctaaaat ggagaaaaaa
     1021 atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca
     1081 tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt
     1141 ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc
     1201 cgcctgatga atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata
     1261 tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg
     1321 ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg
     1381 gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc
     1441 gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac
     1501 aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg
     1561 atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg
     1621 cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc
     1681 agttattggt gcccttaaac gcctggggta atgactctct agcttgaggc atcaaataaa
     1741 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     1801 tctcctgagt aggacaaatc cgccctctag agctgcctcg cgcgtttcgg tgatgacggt
     1861 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     1921 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
     1981 atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc
     2041 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     2101 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2161 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2221 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2281 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2341 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2401 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2461 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2521 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2581 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2641 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2701 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     2761 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2821 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2881 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2941 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3001 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3061 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3121 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3181 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3241 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3301 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3361 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3421 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3481 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3541 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3601 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3661 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3721 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3781 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3841 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3901 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3961 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4021 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     4081 taacctataa aaataggcgt atcacgaggc cctttcgtct tcacctcgag aaa
//
