LOCUS       sequence-374284-        9660 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BiFC plasmid for C-terminal tagging of genes with yEmVenus (aa 
            173–238).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9660)
  AUTHORS   Subotic A, Swinnen E, Demuyser L, De Keersmaecker H, Mizuno H, 
            Tournu H, Van Dijck P
  TITLE     A Bimolecular Fluorescence Complementation Tool for Identification 
            of Protein-Protein Interactions in Candida albicans.
  JOURNAL   G3 (Bethesda). 2017 Oct 5;7(10):3509-3520. doi: 
            10.1534/g3.117.300149.
  PUBMED    28860184
REFERENCE   2  (bases 1 to 9660)
  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: "10.1534/g3.117"; 
            journalName: "G3 (Bethesda)"; date: "2017-10-5- 5"; volume: "7"; 
            issue: "10"; pages: "3509-3520"
COMMENT     Sequence Label: BiFC4
FEATURES             Location/Qualifiers
     source          1..9660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..44
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    161..182
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(197..202,203..220,221..227)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    235..251
                     /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)."
     primer_bind     240..262
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     259..275
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     259..275
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        293..311
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     293..310
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4677..4979
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4858..4877
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             5328..6122
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5328..6122
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5382..5401)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5992..6011
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      6322..6331
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      8945..9533
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9434..9453
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
       61 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
      121 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
      181 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
      241 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctatttaggt
      301 gacactatag aatactcaag ctatgcatca agcttggtac cgagctcgga tccactagta
      361 acggccgcca gtgtgctgga attcgccctt aagaacattt ctgtaccgca aatgtatcga
      421 ctttttcgat ggtgtgttat tgttgttttg ttgcgaaata taatagcttc tgctttatat
      481 cctttgtgtt gatccttccc ctcctttcat acttgtaaaa ttacaattgg tattttcaac
      541 caatattacc acacgtcatg cactaactca aatgggcaaa aaaaagagag ataggaatta
      601 cgtcatttta ttatgttttt acgtggacta gaaggttaag ataatttctg gttattttgt
      661 tgataattga gttttgttcg acctccacca atctttaaac tctcgttgag tattcatttt
      721 aatgattttt tgaaatcaga aggggtgttt ctaacccagg attgttttat tgagaggttg
      781 aaacatcaat gtgttagttt attgatttgg gttggtttaa tagaaaaata tcagtaggtg
      841 ttcaataatt gtagtttatt ttcttggcta aaattgttgc gaaaaattga caaaataata
      901 acaaaaagta ttttagatgc taaaacataa tagtaggaat atctacctga accatttaca
      961 ttcaacaact ggcaagaacc ttcctaataa tttgcaagag taatcacaat taaaccaaaa
     1021 tcaaagttta ccgtgtatat agttggacag aaagctgtca cattggttta atacatagtt
     1081 gaatatgtat gcatgaaaca acaacaatac aattacagct catgatcctt caattaattt
     1141 caacattcaa acaacataac accttgtgta aatgaggtta tgagtacatg tgaacaacaa
     1201 atcttatgac taaactcggg tcgtgtactt ttgacgattg agtcattttt accaattagg
     1261 caattaattt ttgctaaatt ttcattaacc gcgctggcca tcccaacaaa caatataaaa
     1321 gctgacaacg nanaaaaaaa aaaaaaaaaa ctccaattct gaaaaatttt tcttcttctt
     1381 catactaaac aaaacaacta taaagataat caattaataa tgtcacaaca acctagtgaa
     1441 aataaattat ggggtggtcg attcactggc gccactgatc cattaatgga tttatacaat
     1501 gcttcattac cttatgataa agttatgtat gatgcagatt taactggtac taaagtatat
     1561 actcaaggat tgaacaaatt aggattaatt accactgaag aattgaatct gattcatcaa
     1621 ggtttagaac aaattagaca agaatggcat gataataaat ttattattaa agctggtgat
     1681 gaagatattc atactgctaa tgaacgtaga ttaggagaaa tcattgggaa aaacatttct
     1741 ggtaaagttc atactggtag atccagaaat gatcaagttg ctactgatat gagaattttt
     1801 gttcgtgaat cattacttaa tttaagtaag attttacatc aattcattac agcaattctt
     1861 gaacgagctc ataaagaaat tgatgtatta atgcctggtt atactcattt acaaaaagct
     1921 caacctatta gatgggctca ttggttaagt tcatatgcaa cttatttcac tgaagattat
     1981 aaacgtttac aagaaatcat tactcgtgtt aatcaatctc cattaggatc tggtgcttta
     2041 gctggtcatc cctacggtat tgatcgtgaa tttttagcta aaggattagg atttgatggt
     2101 gttattggga attctttaac tgcagtttct gatcgtgatt ttgttgttga aagtttattt
     2161 tggagtacat tatttatgaa tcatatttct cgattttcag aagatttaat tatttattct
     2221 agtggtgaat ttggatttat aaagttagct gatgcttact ctactggttc atcattaatg
     2281 ccacaaaaga aaaatcctga ttcattagaa ttattacgtg ggaaaagtgg cagagtgttt
     2341 ggtcagttat ctgggttttt aatgtcaatt aaatccattc catcaaccta taataaagat
     2401 atgcaagaag ataaagaacc attatttgat gcattgacta ccgtagaaca ttcaatttta
     2461 attgctactg gtgtgatttc tactttactg attgataaac aaaatatgga aaaagcttta
     2521 actatggata tgttagctac tgatttagca gattatttag ttagaaaagg tgttccattt
     2581 agagaaactc atcatatttc tggtgaatgt gttagaaaag ctgaagaaga aaaattgtcg
     2641 ggaattgacc aattatcatt tgaacaattt caacaaattg attcacgttt tgaaaaagat
     2701 gttatggaaa cttttgattt tgaagctagt gtggaaagaa gagatgctat tggtggtact
     2761 gctaagagcg ctgttttaaa acagttagaa aatttgaaat caattttaaa ttaatttata
     2821 aataatcata tagtattcaa tgttttgttt tgtttttttt ttttttttat ggaaatgtac
     2881 aaacagtata taatcaatta atagaaagat accctgtatt ccattgtatc gcctttggga
     2941 tgtctataat accaactttc aagaatgaga aattaaacaa caatttcttt caccaatcaa
     3001 ctcgtctata ttcaataaac tcattctggt cttttcaaat atttaatgtt atcctgatca
     3061 aacttacaaa tcaaaaatgt atcaaaatca ttcattagtt cttcttcttt tgttatcttt
     3121 gtcactatat ttctctccat ccagaccaaa aatacttcca ttgtgaagat cgtctgaatc
     3181 ataatcagca actacctctt ccatttcaga gaattcagat ttttcaaaat aggtatcacg
     3241 ggccaagtac acacaagaca aacgatacga gtgctgggat atttctttca atgtatcact
     3301 aaattcgttg agatttaaag agagcccttt gactgctttc attactttcc tcacatcatg
     3361 agcatccaca ttcaaatttc ctgtagccct ttattcttag ttttcttttc ttattctttt
     3421 cttcgtacat tgaaatcaca aatggtgata tcaaacgaag gtcacactga cttatgtcta
     3481 atacctccaa tgatgatact ggtaacttaa ccaggtctac tcttttcaaa cgatttctca
     3541 ttagtgtcaa tttacgtagg ttcttcaacc tttcaaacga tgcaaatagc ttttgtaagg
     3601 gcgaattctg cagatatcca tcacactggc ggccggccgc tcgagcatgc atctagaggg
     3661 cccttgaaga attggaagaa gttagtagtt aaaaaaacaa tgggaaagat ttaatctttt
     3721 tttctttttt caaaaatttt taaccgctga tgatatatta atctctctcc accactctct
     3781 ttaaacacca caattattaa taataagata tctctcctac tagttatttg ttagtaagta
     3841 tattgttgat tgattggttg ggttaaaaat tttttttttc ataaagaaaa taatgaaagt
     3901 gagaaaaatt taaaccccac attacatact caacacaaca ctacacacac gcatttaaca
     3961 atcatcatca gttgaaatat attttacaaa caattacgat tgtttaatga aaactatttc
     4021 tgttgttatt attatttaac cttttttttc ttgttgggtt gacccttttt ctattaacta
     4081 tctaactacc acataacatt tgaagttgaa ctattaaacc ttaatttgcg gccgcaaatc
     4141 taatctactt tcttcttctt ctaccctatt tattctgaaa tggggggggg ggggtggtgg
     4201 gaactttagg cgtgttaaat ttgtcacatc tgttctcact tgcttctatt cgtataattt
     4261 taattgtctt gtgagatcaa tcttgttctt gttcgagttt tttcttgctt tgattaacta
     4321 ctattattat tagtatgcaa tgctgtttat taccatttca taacttatat tataactcat
     4381 tgatttcaat acctgctcaa caatgagtcg ttattattac tgtgctctgt attctgtact
     4441 gctgcacgta cgttatgtcc attgaagaca agcaaagaaa gaccaacatt caattgtatc
     4501 tgcttaccag agttgtttac ctctgttacc aatatttgga aattaatttt ttggtggtgg
     4561 acgtttcttc ctaaaacacc tagatgtgct agtataattg cacaaccagc acgcacactg
     4621 cgtctggccg gtcctggact acattttgtc tctaacattc gtagctgcac gtacggcagt
     4681 ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg
     4741 atattattga cacgccgggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt
     4801 cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca
     4861 tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc
     4921 tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa
     4981 tgtcaggcat gagattatca aaaaggatct tcacctagat ccttttcacg tagaaagcca
     5041 gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc tggacaaggg
     5101 aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag
     5161 actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta
     5221 aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc
     5281 gcaggggatc aagctctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     5341 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     5401 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     5461 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     5521 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     5581 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     5641 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     5701 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5761 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5821 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     5881 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5941 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     6001 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     6061 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     6121 gaattattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     6181 tttcacaccg catcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     6241 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     6301 caataatagc acgtgaggag ggccaccatg gccaagttga ccagtgccgt tccggtgctc
     6361 accgcgcgcg acgtcgtcta ttccaagcct gtggcttttt ttggtgattc acaatacata
     6421 aattggcaaa tacctcttct atagacttta acctttgatg aaatggtatg gataaaccac
     6481 ttttgtggtc agggtatata tatttttatt ccaatctggc ctttataaat tcaaatttcc
     6541 aaggggactc tgctaaacct tctgatataa ctataactct aataatggct gtaaaaagtg
     6601 tgaaaaaaag atgatattca caaaaaagat aaaggggatt tcatattggg tatctagatt
     6661 taaacatcgt atttcaacaa tgaaactaga atcaatacga aatccagggc acaactgtgt
     6721 tcattccctt gggaattata ctgagtaatg ggaagaatgt gcaaaaaata cccgtgtctt
     6781 atataatttc catttgtgta gggcttaaaa gtggacaact tcaggttacg aaaccacgaa
     6841 aatgtctaaa cgtttgatca aattcgtaac tcatggaaaa tcacagcaac aaattaagga
     6901 tggatatatc cggtacttac aaataaaaac actaaagaag tataaatttt gaactgaaat
     6961 tctatagaaa atcttttttt ttgggtaaaa ttatgcgatt gtggctcata gtaacgtaat
     7021 catacagttc aacaaacatc attttgtcaa aggatttcta atcctattct ttttctactt
     7081 ggcttttgag tatatatata tatatatata catcctttgt tcatcatcat gttcgattga
     7141 atttttttct tggttgttat tagattgtct acttgagttt ttcttggtgc aaattgtggt
     7201 gtttttctaa caaatttttc aacatttaaa gtatttctgg aacttttcta caaggaacga
     7261 catagatcca ttaatgcgcc cctctaaaac aatacccata atgtcttctt aagtatgcat
     7321 ctgcttataa atgtttctgg atttacttta gttagaatgt cgacacgaca atgaactatt
     7381 gctgtagatc acgtgcgatc actgaattat gagtttcttt ttatttttct ttttgggctt
     7441 ttttttttgc aatttttttt ttccaccttt gccaattgcg tagtcacata taatgtatac
     7501 aatttttgca actctgacac tataattgat ccaataataa agtatataat agatcatcaa
     7561 gtatacgtaa tctccccaaa aaaaaaattg cataaaataa aatctataaa ccttttttag
     7621 atttttagtt gttccttttt tctttctatt cttcaacttc caagtgttgt cactttcttc
     7681 ctgttaacta tatttaaaag taaataccct ccccagaaaa ctccggaatg agcctaggga
     7741 cccgggggcg cgccggaggt ggaggttctg gtggaggtgg ttcagctagc gatggtggtg
     7801 ttcaattagc tgaccattat caacaaaata ctccaattgg tgatggtcca gtcttgttac
     7861 cagacaacca ttacttatcc tatcaatcta aattatccaa agatccaaac gaaaagagag
     7921 accacatggt cttgttagaa tttgttactg ctgctggtat tacccatggt atggatgaat
     7981 tgtacaaata aacgcgtagt acttaagatc ctgaagacgg gatcgttcga agagtgaaat
     8041 tctggaaatc tggaaatctg gttttgtatt cttgttattc ttctttttgt tattacatat
     8101 ataacttgtt acttttttaa aaaaatcttt gtttatttta taaatatata aaactaaatt
     8161 taagaaaaag agaaaaatgt tttatttgag agattgaaat tttacttgaa tttagcttag
     8221 cttttataaa gtattattat gtaaaaaaac aaaacaaata tacattaaaa agttaagact
     8281 ataaaatagc cacccaaggc atttctatat cttgttgttg ttgttttcat cttctgtatc
     8341 agaggaactt attttattat tttcgtcacg ggtattttct cttgtttgat gattcatccc
     8401 attcattcca tcataaaatt ctagaaggac cacctttgat tgtaaatagt aataattacc
     8461 acccttattt aattatttat ttaacttatt tatttattta ttatacatat atacaaatct
     8521 aataaagtga aaatctcccc cttcacaccg gtgtggtccg ggacgacgtg accctgttca
     8581 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     8641 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     8701 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     8761 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctaaaac
     8821 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     8881 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     8941 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     9001 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     9061 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     9121 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     9181 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     9241 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     9301 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     9361 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     9421 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     9481 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     9541 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     9601 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
//
