LOCUS       sequence-377523-        9060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BiFC plasmid for N-terminal tagging of genes with yEmVenus (aa 
            1–172).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9060)
  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 9060)
  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: BiFC1
FEATURES             Location/Qualifiers
     source          1..9060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    90..111
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(126..131,132..149,150..156)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    164..180
                     /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     169..191
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     188..204
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     188..204
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        222..240
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     222..239
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2578..2597)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2579..2597)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2604..2621)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2604..2620)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2613..2635)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             3847..4149
                     /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     4028..4047
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             4498..5292
                     /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             4498..5292
                     /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(4552..4571)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5162..5181
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      5492..5501
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      8274..8862
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8763..8782
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9016..9033
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
       61 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
      121 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
      181 tttcacacag gaaacagcta tgaccatgat tacgccaagc tatttaggtg acactataga
      241 atactcaagc tatgcatcaa gcttggtacc gagctcggat ccactagtaa cggccgccag
      301 tgtgctggaa ttcgcccttg gatccaatca tcactggtgg gggtcaagaa agaggtttga
      361 gatcaggtac tttggcacca ccattagtag ctgggtttgg tgaagctgca agattaatga
      421 aacaagaatc tgcatttgac aagaaacata ttgagaggtt atctactaaa ttgaagaacg
      481 gtttgttatc cattccatct actcagttta acggttgtaa taacccaaca tatcaatacc
      541 caggttgtgt gaatgtttcc tttgcataca ttgaagggga atctttgtta atggcattga
      601 aagatattgc attgagttct ggttctgcat gtacctctgc atctttggaa ccatcgtatg
      661 ttttacatgc tttgggtgcc gatgatgcct tggctcattc ttccattaga ttcggtattg
      721 gtagattcac cactgaagca gaagttgact atgttattca agcaattaat gaaagagttg
      781 atttcttgag aaagatgtct ccattatggg aaatggttca agaaggtatt gacttgaact
      841 ctatcgaatg gagtggacat taaggttttt tgtttttttg taatttgtgt tgttatgtgt
      901 aattatcgtg tatagagtaa tttcggaaat tgtactttcg ttcatgtatc tgaaccagat
      961 gaataatacg ttcgtattgt tgaagaatga ctaactcctg taacggcgtg cgtgggttcc
     1021 gcagccgggt tcaatttttt ttttcttgtt cactataaaa atatataata aacgaaaaaa
     1081 atccgaggga ctcaggctat gaaatttttc actcaagaat tttttttcct tcatccttaa
     1141 ttcctttatt cactacccaa acccacaatg tcagttaaaa ctaaaactat tacaatttta
     1201 ccaggtgatc acgtcggtac cgaaattgtc aatgaagcta tcaaagtttt agaagcaatt
     1261 gaagctgcca ctccttacca aaaaatccat ttcgatttca aacatcattt aatcggtggt
     1321 gctgccattg atgccactgg tgtcccattg ccagatgacg cacttgaaag tgccaaaaaa
     1381 tctgatgccg tcttattggg tgctgttggt ggaccaaaat gggggactgg cgccgttcgt
     1441 ccagaacaag gtttattaaa gattcgtaaa gaattgaacc tttatgctaa catcagacca
     1501 tgtaactttg ccagtgactc cttattggaa ttatccccat tgagaccaga agtcgttaaa
     1561 ggtacaaact tgatcattgt tcgtgaatta gttggtggta tttatttcgg tgatcgtgaa
     1621 gaacaagaag aaagtgaaga taaacaaacc gcatgggata ctgaaaaata caccgtcgat
     1681 gaagtcacca gaatcacccg tatggctgcc ttcatggctt tacaacataa cccaccatta
     1741 ccaatctggt cattggataa ggctaatgtt ttagcttctt ctagattatg gagaagaact
     1801 gtcgataaag tgatttctga agagttccca actttgtctg ttcaacatca attgattgac
     1861 tccgccgcca tgattttaat tcaaaaccca accaaattga atggtataat catcacttct
     1921 aacatgtttg gtgatatcat ttccgatgaa gcttcagtta ttccaggttc cttgggttta
     1981 ttaccatctg catctttggc ttcattgcca gataccaaca ccgcttttgg tctttatgaa
     2041 ccatgtcacg gttctgctcc tgatttacca gccaataaag ttaacccaat tgctactatt
     2101 ttatctgctg cttctatgtt gagattatct ttggattgtg tgaaagaagc tgaagcttta
     2161 gaagaagccg tgaaacaagt cttggataaa ggtatcagaa ctgcagattt aagaggtacc
     2221 agttcagcta ctgaagttgg tgacgcgatt gtcgaagctg ttactaaaat cttaaaagaa
     2281 aaagcttaag taaagtatac acaaaattta acgaatatta ttaatacaaa atgaacacca
     2341 cctaaatact tttttttaaa aatgtaccga ctcttctaaa aattgcgata attcaatatc
     2401 ggaaatttct ctcaccggtt tacttggatc ttcggaaata aatctatttg aagtaatggt
     2461 agcctcactt gattctgatt ggctctttct agacatgggt aggtaagggc gaattctgca
     2521 gatatccatc acactggcgg ccggccgctc gagcatgcat ctagagggcc caattcgccc
     2581 tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     2641 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     2701 agcgaagagg cccgcaccga tcgaccagat caaaatgcta gaacaaatta cgtctgcatc
     2761 aacagtcatc ttccttattt gcataaatgc taaaatggca aacctgtaaa ccgtttgaaa
     2821 agcaaattga gatagttagc tcggtttttt tttcacgtgg ccataatcca acaaaaaaaa
     2881 ggaatttaag gggggagaga aggaggcttg aatttgtacc acagagcgag caagaaatta
     2941 ataatatact tggcttagct gaaatgagaa tgaaaaagca aatttcaagt gaaacaccac
     3001 tactagccaa gtaaattggc caatttgctc tcgtgcttta atttcctaac aagcttttga
     3061 gaccgaaaaa aatcaaccaa tgatgatgga aaaatacttc cgggagtcaa tttatccata
     3121 atttcacgtc gttttttttt tgcacatggc aaatatattg gaatcctgag attttcttgg
     3181 agtcgtgtac aataacacca ggcgggaaca tgccgttatg atccagcata gcggccgcta
     3241 tgtgttcaaa tattgaaacc gtgaaaccag tttgatacag ctgatattgt gtgctaaaaa
     3301 aagatgagca ccgaaagagt ctttagttta ttgccaagta aagggtcaaa gaatacctat
     3361 tctttatgaa cacagacttg ctctaatggt gaattagtgt ggagtaatag agaaggaggc
     3421 gaaacaaaaa taattggtta aacactcgaa tgtcacacaa tcctaaatag gttaaattca
     3481 gttaccgccg tgatgcggga acaaagccga gcggtaatat ttttggattc tatgtttttt
     3541 tggtgtatat tgcacgactt ggtgccaaca actggtgttc caacgcacaa aactaggttc
     3601 aactggatac tgttttctag aaattatttg cggcaattca tgattacggt cctaaatttg
     3661 aaataaaagt ttccaattat gtgtcttgac aatttccatt gggttcaaaa agatctagaa
     3721 tgtatgtaat tacgcaacaa tttgcatgca aagaaataaa aacaaaccta aaatccaact
     3781 gctgttgttc tggtgatgat gtagtaaaat aagagcacgg ccaagaacaa ggtgatttac
     3841 gtacggcagt ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat
     3901 gtacagagtg atattattga cacgccgggg cgacggatgg tgatccccct ggccagtgca
     3961 cgtctgctgt cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa
     4021 agctggcgca tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa
     4081 gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg
     4141 ggaatataaa tgtcaggcat gagattatca aaaaggatct tcacctagat ccttttcacg
     4201 tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc
     4261 tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg
     4321 cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg
     4381 ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg
     4441 atctgatggc gcaggggatc aagctctgat caagagacag gatgaggatc gtttcgcatg
     4501 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4561 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4621 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     4681 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4741 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4801 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4861 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4921 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4981 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     5041 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5101 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5161 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5221 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5281 gagttcttct gaattattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat
     5341 ctgtgcggta tttcacaccg catcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5401 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5461 ataaatgctt caataatagc acgtgaggag ggccaccatg gccaagttga ccagtgccgt
     5521 tccggtgctc accgcgcgcg acgtcttcca attagaggtt gtggatgcaa aattgaaaaa
     5581 aaaatgacta ttggtagcgt gttctatttt tcttatgcgc tgaaaaacta cgaacaattg
     5641 tctattccaa gcctgtggct ttttttggtg attcacaata cataaattgg caaatacctc
     5701 ttctatagac tttaaccttt gatgaaatgg tatggataaa ccacttttgt ggtcagggta
     5761 tatatatttt tattccaatc tggcctttat aaattcaaat ttccaagggg actctgctaa
     5821 accttctgat ataactataa ctctaataat ggctgtaaaa agtgtgaaaa aaagatgata
     5881 ttcacaaaaa agataaaggg gatttcatat tgggtatcta gatttaaaca tcgtatttca
     5941 acaatgaaac tagaatcaat acgaaatcca gggcacaact gtgttcattc ccttgggaat
     6001 tatactgagt aatgggaaga atgtgcaaaa aatacccgtg tcttatataa tttccatttg
     6061 tgtagggctt aaaagtggac aacttcaggt tacgaaacca cgaaaatgtc taaacgtttg
     6121 atcaaattcg taactcatgg aaaatcacag caacaaatta aggatggata tatccggtac
     6181 ttacaaataa aaacactaaa gaagtataaa ttttgaactg aaattctata gaaaatcttt
     6241 ttttttgggt aaaattatgc gattgtggct catagtaacg taatcataca gttcaacaaa
     6301 catcattttg tcaaaggatt tctaatccta ttctttttct acttggcttt tgagtatata
     6361 tatatatata tatacatcct ttgttcatca tcatgttcga ttgaattttt ttcttggttg
     6421 ttattagatt gtctacttga gtttttcttg gtgcaaattg tggtgttttt ctaacaaatt
     6481 tttcaacatt taaagtattt ctggaacttt tctacaagga acgacataga tccattaatg
     6541 cgcccctcta aaacaatacc cataatgtct tcttaagtat gcatctgctt ataaatgttt
     6601 ctggatttac tttagttaga atgtcgacac gacaatgaac tattgctgta gatcacgtgc
     6661 gatcactgaa ttatgagttt ctttttattt ttctttttgg gctttttttt ttgcaatttt
     6721 ttttttccac ctttgccaat tgcgtagtca catataatgt atacaatttt tgcaactctg
     6781 acactataat tgatccaata ataaagtata taatagatca tcaagtatac gtaatctccc
     6841 caaaaaaaaa attgcataaa ataaaatcta taaacctttt ttagattttt agttgttcct
     6901 tttttctttc tattcttcaa cttccaagtg ttgtcacttt cttcctgtta actatattta
     6961 aaagtaaata ccctccccag aaaactccgg aatgtctaaa ggtgaagaat tattcactgg
     7021 tgttgtccca attttggttg aattagatgg tgatgttaat ggtcacaaat tttctgtctc
     7081 cggtgaaggt gaaggtgatg ctacttacgg taaattgacc ttaaaattga tttgtactac
     7141 tggtaaattg ccagttccat ggccaacctt agtcactact ttaggttatg gtttgcaatg
     7201 ttttgctaga tacccagatc atatgaaaca acatgacttt ttcaagtctg ccatgccaga
     7261 aggttatgtt caagaaagaa ctattttttt caaagatgac ggtaactaca agaccagagc
     7321 tgaagtcaag tttgaaggtg ataccttagt taatagaatc gaattaaaag gtattgattt
     7381 taaagaagat ggtaacattt taggtcacaa attggaatac aactataact ctcacaatgt
     7441 ttacatcact gctgacaaac aaaagaatgg tatcaaagct aacttcaaaa ttagacacaa
     7501 cattgaagct agcggaggtg gaggttctgg tggaggtggt tcaagcctag ggacccgggg
     7561 gcgcgcctaa acgcgtagta cttaaccgcg ccgtcgacga gtgaaattct ggaaatctgg
     7621 aaatctggtt ttgtattctt gttattcttc tttttgttat tacatatata acttgttact
     7681 tttttaaaaa aatctttgtt tattttataa atatataaaa ctaaatttaa gaaaaagaga
     7741 aaaatgtttt atttgagaga ttgaaatttt acttgaattt agcttagctt ttataaagta
     7801 ttattatgta aaaaaacaaa acaaatatac attaaaaagt taagactata aaatagccac
     7861 ccaaggcatt tctatatctt gttgttgttg ttttcatctt ctgtatcaga ggaacttatt
     7921 ttattatttt cgtcacgggt attttctctt gtttgatgat tcatcccatt cattccatca
     7981 taaaattcta gaaggaccac ctttgattgt aaatagtaat aattaccacc cttatttaat
     8041 tatttattta acttatttat ttatttatta tacatatata caaatctaat aaagtgaaaa
     8101 tctccccctt cacacgtggc cgaggagcag gactgacacg tgctaaaact tcatttttaa
     8161 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     8221 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     8281 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     8341 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     8401 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     8461 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     8521 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     8581 cggtcgggct gaacgggggg gtcgtgcaca cagcccagct tggagcgaac gacctacacc
     8641 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     8701 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     8761 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     8821 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     8881 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     8941 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     9001 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
//
