LOCUS       sequence-176199-        6536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FKBP-tagged Dronpa..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6536)
  AUTHORS   Mo GC, Ross B, Hertel F, Manna P, Yang X, Greenwald E, Booth C, 
            Plummer AM, Tenner B, Chen Z, Wang Y, Kennedy EJ, Cole PA, Fleming 
            KG, Palmer A, Jimenez R, Xiao J, Dedecker P, Zhang J
  TITLE     Genetically encoded biosensors for visualizing live-cell biochemical
            activity at super-resolution.
  JOURNAL   Nat Methods. 2017 Apr;14(4):427-434. doi: 10.1038/nmeth.4221. Epub 
            2017 Mar 13.
  PUBMED    28288122
REFERENCE   2  (bases 1 to 6536)
  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: 
            "10.1038/nmeth.4221"; journalName: "Nat Methods"; date: "2017-04"; 
            volume: "14"; issue: "4"; pages: "427-434"
COMMENT     Sequence Label: FKBP-Dronpa
FEATURES             Location/Qualifiers
     source          1..6536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..18
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             22..54
                     /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(58..66,67..81)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(88..90,91..411)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             433..1107
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVK
                     EGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNAT
                     NDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALSL
                     EGGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPRQ
                     AK"
     promoter        complement(1168..1186)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1169..1186)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1206..1223)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1212..1436
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1482..1910
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1569..1588)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1779..1800
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1919..1939)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1924..2253
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2104..2239
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2166..2185
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2320..3114
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2374..2393)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2984..3003
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3288..3409
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3325..3344)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3379..3398
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3458..3474)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3458..3474)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3471..3493)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3482..3498
                     /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)."
     promoter        complement(join(3506..3512,3513..3530,3531..3536))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3551..3572
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3689..3706)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3860..4448)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3940..3959)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4619..5410,5411..5479))
                     /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     5242..5261
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5480..5584)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5659..5678)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5850..6229
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6230..6433
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6384..6404
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6478..6497
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6478..6496
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6516..6525
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 catcatcatc atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat
       61 ctgtacgacg atgacgataa ggatccgatg ggagtgcagg tggaaaccat ctccccagga
      121 gacgggcgca ccttccccaa gcgcggccag acctgcgtgg tgcactacac cgggatgctt
      181 gaagatggaa agaaatttga ttcctcccgg gacagaaaca agccctttaa gtttatgcta
      241 ggcaagcagg aggtgatccg aggctgggaa gaaggggttg cccagatgag tgtgggtcag
      301 agagccaaac tgactatatc tccagattat gcctatggtg ccactgggca cccaggcatc
      361 atcccaccac atgccactct cgtcttcgat gtggagcttc taaaactgga aggtaccgga
      421 ggtagcggtt ctatgagtgt gattaaacca gacatgaaga tcaagctgcg tatggaaggc
      481 gctgtaaatg gacacccgtt cgcgattgaa ggagttggcc ttgggaagcc tttcgaggga
      541 aaacagagta tggaccttaa agtcaaagaa ggcggacctc tgcctttcgc ctatgacatc
      601 ttgacaactg tgttctgtta cggcaacagg gtattcgcca aatacccaga aaatatagta
      661 gactatttca agcagtcgtt tcctgagggc tactcttggg aacgaagcat gaattacgaa
      721 gacgggggca tttgtaacgc gacaaacgac ataaccctgg atggtgactg ttatatctat
      781 gaaattcgat ttgatggtgt gaactttcct gccaatggtc cagttatgca gaagaggact
      841 gtgaaatggg agccatccac tgagaaattg tatgtgcgtg atggagtgct gaagggtgat
      901 gttaacatgg ctctgtcgct tgaaggaggt ggccattacc gatgtgactt caaaactact
      961 tataaagcta agaaggttgt ccagttgcca gactatcact ttgtggacca ccacattgag
     1021 attaaaagcc acgacaaaga ttacagtaat gttaatctgc atgagcatgc cgaagcgcat
     1081 tctgagctgc cgaggcaggc caagtaagaa ttctgcagat atccatcaca ctggcggccg
     1141 ctcgagcatg catctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg
     1201 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     1261 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     1321 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     1381 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     1441 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     1501 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     1561 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     1621 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     1681 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     1741 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     1801 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     1861 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     1921 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     1981 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2041 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     2101 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     2161 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     2221 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     2281 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     2341 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     2401 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     2461 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     2521 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     2581 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     2641 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     2701 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     2761 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     2821 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     2881 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     2941 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     3001 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     3061 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     3121 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     3181 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     3241 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     3301 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3361 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     3421 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     3481 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     3541 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     3601 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3661 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3721 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3781 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3841 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3901 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3961 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4021 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4081 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4141 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4201 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4261 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4321 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4381 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4441 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4501 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4561 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4621 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4681 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4741 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4801 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4861 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4921 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4981 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5041 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5101 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5161 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5221 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5281 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5341 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5401 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5461 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5521 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5581 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     5641 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     5701 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     5761 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     5821 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
     5881 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     5941 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     6001 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     6061 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     6121 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     6181 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     6241 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     6301 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     6361 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     6421 tataagcaga gctctctggc taactagaga acccactgct tactggctta tcgaaattaa
     6481 tacgactcac tatagggaga cccaagcttg cggccgccac catggtgcgg ggttct
//
