LOCUS       sequence-176120-        6311 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRB-tagged N-terminal Dronpa fragment for BiFC; localizes to plasma 
            membrane..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6311)
  AUTHORS   Hertel F, Mo GC, Duwe S, Dedecker P, Zhang J
  TITLE     RefSOFI for Mapping Nanoscale Organization of Protein-Protein 
            Interactions in Living Cells.
  JOURNAL   Cell Rep. 2016 Jan 12;14(2):390-400. doi: 
            10.1016/j.celrep.2015.12.036. Epub 2015 Dec 31.
  PUBMED    26748717
REFERENCE   2  (bases 1 to 6311)
  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: 
            "10.1016/j.celrep.2015.12.036"; journalName: "Cell Rep"; date: 
            "2016-01-12- 12"; volume: "14"; issue: "2"; pages: "390-400"
COMMENT     Sequence Label: pcDNA3-Lyn-FRB-DMVF-181-N
FEATURES             Location/Qualifiers
     source          1..6311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             40..318
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     promoter        complement(958..976)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(959..976)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(996..1013)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1002..1226
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1272..1700
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1359..1378)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1569..1590
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1709..1729)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1714..2043
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1894..2029
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1956..1975
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2110..2904
                     /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(2164..2183)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2774..2793
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3078..3199
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3115..3134)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3169..3188
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3248..3264)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3248..3264)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3261..3283)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3272..3288
                     /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(3296..3302,3303..3320,3321..3326))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3341..3362
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3479..3496)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3650..4238)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3730..3749)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4409..5200,5201..5269))
                     /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     5032..5051
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5270..5374)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5449..5468)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5640..6019
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6020..6223
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6174..6194
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6268..6287
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6268..6286
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      join(6306..6311,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atgggctgca tcaagagcaa gcgcaaggac aaggatccca tcctctggca tgagatgtgg
       61 catgaaggcc tggaagaggc atctcgtttg tactttgggg aaaggaacgt gaaaggcatg
      121 tttgaggtgc tggagccctt gcatgctatg atggaacggg gcccccagac tctgaaggaa
      181 acatccttta atcaggccta tggtcgagat ttaatggagg cccaagagtg gtgcaggaag
      241 tacatgaaat cagggaatgt caaggacctc ctccaagcct gggacctcta ttatcatgtg
      301 ttccgacgaa tctcaaagac tagtggtacc ggtggaggtg gatctggagg aatgagtgtg
      361 attaaaccag acatgaagat caagctgcgt atggaaggcg ctgtaaatgg acacccgttc
      421 gcgattgaag gagttggcct tgggaagcct ttcgagggaa aacagagtat agaccttaaa
      481 gtcaaagaag gcggacctct gcctttcgcc tatgacatct tgacaactgc gttctgttac
      541 ggcaacaggg tattcgccaa atacccagaa aatatagtag actatttcaa gcagtcgttt
      601 cctgagggct actcttggga acgaagcatg aattacgaag acgggggcat ttgtaacgcg
      661 acaaacgaca taaccctgga tggtgactgt tatatctatg aaattcgatt tgatggtgtg
      721 aactttcctg ccaatggtcc agttatgcag aagaggactg tgaaatggga gccatccact
      781 gagaaattgt atgtgcgtga tggagtgctg aagggtgatg ttaacatggc tctgtcgctt
      841 gaaggaggtg gccattaccg atgtgacgtt aaaactactt ataaagctaa gaagtaagaa
      901 ttctgcagat atccatcaca ctggcggccg ctcgagcatg catctagagg gccctattct
      961 atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
     1021 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     1081 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     1141 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     1201 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     1261 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     1321 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     1381 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     1441 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     1501 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     1561 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     1621 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     1681 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     1741 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     1801 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     1861 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     1921 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     1981 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     2041 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     2101 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     2161 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     2221 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     2281 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     2341 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     2401 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     2461 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     2521 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     2581 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     2641 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     2701 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     2761 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     2821 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     2881 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     2941 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     3001 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     3061 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3121 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3181 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     3241 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3301 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3361 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3421 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3481 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3541 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3601 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3661 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3721 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3781 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3841 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3901 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3961 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4021 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4081 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4141 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4201 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4261 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4321 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4381 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4441 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4501 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4561 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4621 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4681 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4741 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4801 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4861 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4921 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4981 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5041 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5101 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5161 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5221 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5281 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5341 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     5401 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
     5461 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     5521 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     5581 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
     5641 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     5701 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     5761 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     5821 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     5881 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     5941 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     6001 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     6061 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     6121 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     6181 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
     6241 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagcttg
     6301 cggccgccac c
//
