LOCUS       sequence-176114-        5999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FKBP-tagged C-terminal Dronpa fragment for BiFC..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5999)
  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 5999)
  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-FKBP-DMVF-181-C
FEATURES             Location/Qualifiers
     source          1..5999
                     /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"
     promoter        complement(631..649)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(632..649)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(669..686)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    675..899
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      945..1373
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1032..1051)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1242..1263
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1382..1402)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1387..1716
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1567..1702
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1629..1648
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1783..2577
                     /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(1837..1856)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2447..2466
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2751..2872
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2788..2807)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2842..2861
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2921..2937)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2921..2937)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2934..2956)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2945..2961
                     /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(2969..2975,2976..2993,2994..2999))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3014..3035
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3152..3169)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3323..3911)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3403..3422)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4082..4873,4874..4942))
                     /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     4705..4724
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4943..5047)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5122..5141)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5313..5692
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5693..5896
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5847..5867
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5941..5960
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5941..5959
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5979..5988
                     /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 ggtagcggag gtggaagtgt tgtccagttg ccagactatc actttgtgga ccaccacatt
      481 gagattaaaa gccacgacaa agattacagt aatgttaatc tgcatgagca tgccgaagcg
      541 cattctgagc tgccgaggca ggccaagtaa gaattctgca gatatccatc acactggcgg
      601 ccgctcgagc atgcatctag agggccctat tctatagtgt cacctaaatg ctagagctcg
      661 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
      721 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
      781 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
      841 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
      901 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
      961 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1021 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     1081 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1141 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     1201 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     1261 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     1321 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     1381 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     1441 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     1501 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     1561 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     1621 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     1681 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     1741 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     1801 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     1861 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     1921 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     1981 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     2041 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     2101 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     2161 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     2221 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     2281 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     2341 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     2401 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     2461 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     2521 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     2581 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     2641 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     2701 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     2761 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2821 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     2881 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     2941 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3001 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3061 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3121 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3181 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3241 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3301 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3361 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3421 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3481 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3541 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3601 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3661 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3721 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3781 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3841 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3901 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     3961 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4021 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4081 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4141 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4201 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4261 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4321 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4381 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4441 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4501 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4561 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4621 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4681 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4741 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4801 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4861 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4921 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4981 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5041 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     5101 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     5161 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     5221 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     5281 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
     5341 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     5401 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     5461 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     5521 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     5581 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     5641 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     5701 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     5761 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     5821 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     5881 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
     5941 taatacgact cactataggg agacccaagc ttgcggccgc caccatggtg cggggttct
//
