LOCUS       sequence-367926-        9743 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 9743)
  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 9743)
  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..9743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     239..256
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4612..4914
                     /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     4793..4812
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             5263..6057
                     /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             5263..6057
                     /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(5317..5336)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5927..5946
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      6258..6267
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      8974..9562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9463..9482
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9716..9733
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagtttctg taccgcaaat gtatcgactt tttcgatggt gtgttattgt tgttttgttg
      361 cgaaatataa tagcttctgc tttatatcct ttgtgttgat ccttcccctc ctttcatact
      421 tgtaaaatta caattggtat tttcaaccaa tattaccaca cgtcatgcac taactcaaat
      481 gggcaaaaaa aagagagata ggaattacgt cattttatta tgtttttacg tggactagaa
      541 ggttaagata atttctggtt attttgttga taattgagtt ttgttcgacc tccaccaatc
      601 tttaaactct cgttgagtat tcattttaat gattttttga aatcagaagg ggtgtttcta
      661 acccaggatt gttttattga gaggttgaaa catcaatgtg ttagtttatt gatttgggtt
      721 ggtttaatag aaaaatatca gtaggtgttc aataattgta gtttattttc ttggctaaaa
      781 ttgttgcgaa aaattgacaa aataataaca aaaagtattt tagatgctaa aacataatag
      841 taggaatatc tacctgaacc atttacattc aacaactggc aagaaccttc ctaataattt
      901 gcaagagtaa tcacaattaa accaaaatca aagtttaccg tgtatatagt tggacagaaa
      961 gctgtcacat tggtttaata catagttgaa tatgtatgca tgaaacaaca acaatacaat
     1021 tacagctcat gatccttcaa ttaatttcaa cattcaaaca acataacacc ttgtgtaaat
     1081 gaggttatga gtacatgtga acaacaaatc ttatgactaa actcgggtcg tgtacttttg
     1141 acgattgagt catttttacc aattaggcaa ttaatttttg ctaaattttc attaaccgcg
     1201 ctggccatcc caacaaacaa tataaaagct gacaacgaaa aaaaaaaaaa aaaaaaaaaa
     1261 actccaattc tgaaaaattt ttcttcttct tcatactaaa caaaacaact ataaagataa
     1321 tcaattaata atgtcacaac aacctagtga aaataaatta tggggtggtc gattcactgg
     1381 cgccactgat ccattaatgg atttatacaa tgcttcatta ccttatgata aagttatgta
     1441 tgatgcagat ttaactggta ctaaagtata tactcaagga ttgaacaaat taggattaat
     1501 taccactgaa gaattgaatc tgattcatca aggtttagaa caaattagac aagaatggca
     1561 tgataataaa tttattatta aagctggtga tgaagatatt catactgcta atgaacgtag
     1621 attaggagaa atcattggga aaaacatttc tggtaaagtt catactggta gatccagaaa
     1681 tgatcaagtt gctactgata tgagaatttt tgttcgtgaa tcattactta atttaagtaa
     1741 gattttacat caattcatta cagcaattct tgaacgagct cataaagaaa ttgatgtatt
     1801 aatgcctggt tatactcatt tacaaaaagc tcaacctatt agatgggctc attggttaag
     1861 ttcatatgca acttatttca ctgaagatta taaacgttta caagaaatca ttactcgtgt
     1921 taatcaatct ccattaggat ctggtgcttt agctggtcat ccctacggta ttgatcgtga
     1981 atttttagct aaaggattag gatttgatgg tgttattggg aattctttaa ctgcagtttc
     2041 tgatcgtgat tttgttgttg aaagtttatt ttggagtaca ttatttatga atcatatttc
     2101 tcgattttca gaagatttaa ttatttattc tagtggtgaa tttggattta taaagttagc
     2161 tgatgcttac tctactggtt catcattaat gccacaaaag aaaaatcctg attcattaga
     2221 attattacgt gggaaaagtg gcagagtgtt tggtcagtta tctgggtttt taatgtcaat
     2281 taaatccatt ccatcaacct ataataaaga tatgcaagaa gataaagaac cattatttga
     2341 tgcattgact accgtagaac attcaatttt aattgctact ggtgtgattt ctactttact
     2401 gattgataaa caaaatatgg aaaaagcttt aactatggat atgttagcta ctgatttagc
     2461 agattattta gttagaaaag gtgttccatt tagagaaact catcatattt ctggtgaatg
     2521 tgttagaaaa gctgaagaag aaaaattgtc gggaattgac caattatcat ttgaacaatt
     2581 tcaacaaatt gattcacgtt ttgaaaaaga tgttatggaa acttttgatt ttgaagctag
     2641 tgtggaaaga agagatgcta ttggtggtac tgctaagagc gctgttttaa aacagttaga
     2701 aaatttgaaa tcaattttaa attaatttat aaataatcat atagtattca atgttttgtt
     2761 ttgttttttt tttttttttt atggaaatgt acaaacagta tataatcaat taatagaaag
     2821 ataccctgta ttccattgta tcgcctttgg gatgtctata ataccaactt tcaagaatga
     2881 gaaattaaac aacaatttct ttcaccaatc aactcgtcta tattcaataa actcattctg
     2941 gtcttttcaa atatttaatg ttatcctgat caaacttaca aatcaaaaat gtatcaaaat
     3001 cattcattag ttcttcttct tttgttatct ttgtcactat atttctctcc atccagacca
     3061 aaaatacttc cattgtgaag atcgtctgaa tcataatcag caactacctc ttccatttca
     3121 gagaattcag atttttcaaa ataggtatca cgggccaagt acacacaaga caaacgatac
     3181 gagtgctggg atatttcttt caatgtatca ctaaattcgt tgagatttaa agagagccct
     3241 ttgactgctt tcattacttt cctcacatca tgagcatcca cattcaaatt tcctgtagcc
     3301 ctttattctt agttttcttt tcttattctt ttcttcgtac attgaaatca caaatggtga
     3361 tatcaaacga aggtcacact gacttatgtc taatacctcc aatgatgata ctggtaactt
     3421 aaccaggtct actcttttca aacgatttct cattagtgtc aatttacgta ggttcttcaa
     3481 cctttcaaac gatgcaaata gcttttgtaa cggccgccag tgtgctggaa ttgcccttaa
     3541 gggcgaattc tgcagatatc catcacactg gcggccggcc gctcgagcat gcatctagag
     3601 ggcccttgaa gaattggaag aagttggcag ttaaaaaaca atgggaaaga tttaatcttt
     3661 tttttcttct ttcaaaaatt tttaaccgct gatgatatat taatctctct ccaccactct
     3721 cgttaaacac cacaattatt aataataaga tatctctcct actaaatatt tgttagtaag
     3781 tatattgttg attgattggt tgggttaaaa attttttttt tcataaagaa aataatgaaa
     3841 gtgggaaaaa tttaaacccc acattacata ctcaacacaa cactacacac acgcatttaa
     3901 caatcatcat cagttggaat atattttaca aacaattacg attgtttaat gaaaactatt
     3961 tctgttatta ttatttaacc tttttttttt cttgttgggt tgaccctttt tctattaact
     4021 atctaactac cacataacat ttgaagttga actattgaac cttaatttgc ggccgcaaat
     4081 ttaatcttct ttcttcttcc accttttttt tatttttttg aaatgggggg tggtgggaac
     4141 tttaggcgtg ttaaatttgt cacatctgtt tcacttgctt ctattcgtat aattttaatt
     4201 gtcttgtgag atcaatcttg ttcttgctct tgttttttct tgctttgatt aactaccatt
     4261 attattagta tgcaatgctg tttattacca tttcataact tatattataa ctcattgatt
     4321 tcaatacctg ctcaacaatg agtcgttatt attactgtgc tctgtattct gtactgctgc
     4381 acgtacgtta tgtccattga agacaagcaa agaaagacca acattcaatt gtatctgctt
     4441 accagagttg tttacctctg ttaccaatat ttggaaatta attttttggt ggtggacgtt
     4501 tcttcctaaa acacctagat gtgctagtat aattgcacaa ccagcacgca cactgcgtct
     4561 ggccggtcct ggactacatt ttgtctctaa cattcgtagc tgcacgtacg gcagtttaag
     4621 gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt
     4681 attgacacgc cggggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat
     4741 aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg
     4801 accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc
     4861 caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca
     4921 ggcatgagat tatcaaaaag gatcttcacc tagatccttt tcacgtagaa agccagtccg
     4981 cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac aagggaaaac
     5041 gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata gctagactgg
     5101 gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt
     5161 gggaagccct gcaaagtaaa ctggatggct ttctcgccgc caaggatctg atggcgcagg
     5221 ggatcaagct ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     5281 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5341 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5401 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     5461 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5521 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5581 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5641 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5701 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5761 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     5821 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5881 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5941 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     6001 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgaatt
     6061 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     6121 caccgcatac aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6181 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6241 aatagcacgt gaggagggcc accatggcca agttgaccag tgccgttccg gtgctcaccg
     6301 cgcgcgacgt cttccaatta gaggttgtgg atgcaaaatt gaaaaaaaaa tgactattgg
     6361 tagcgtgttc tatttttctt atgcgctgaa aaactacgaa caattgtcta ttccaagcct
     6421 gtggcttttt ttggtgattc acaatacata aattggcaaa tacctcttct atagacttta
     6481 acctttgatg aaatggtatg gataaaccac ttttgtggtc agggtatata tatttttatt
     6541 ccaatctggc ctttataaat tcaaatttcc aaggggactc tgctaaacct tctgatataa
     6601 ctataactct aataatggct gtaaaaagtg tgaaaaaaag atgatattca caaaaaagat
     6661 aaaggggatt tcatattggg tatctagatt taaacatcgt atttcaacaa tgaaactaga
     6721 atcaatacga aatccagggc acaactgtgt tcattccctt gggaattata ctgagtaatg
     6781 ggaagaatgt gcaaaaaata cccgtgtctt atataatttc catttgtgta gggcttaaaa
     6841 gtggacaact tcaggttacg aaaccacgaa aatgtctaaa cgtttgatca aattcgtaac
     6901 tcatggaaaa tcacagcaac aaattaagga tggatatatc cggtacttac aaataaaaac
     6961 actaaagaag tataaatttt gaactgaaat tctatagaaa atcttttttt ttgggtaaaa
     7021 ttatgcgatt gtggctcata gtaacgtaat catacagttc aacaaacatc attttgtcaa
     7081 aggatttcta atcctattct ttttctactt ggcttttgag tatatatata tatatatata
     7141 catcctttgt tcatcatcat gttcgattga atttttttct tggttgttat tagattgtct
     7201 acttgagttt ttcttggtgc aaattgtggt gtttttctaa caaatttttc aacatttaaa
     7261 gtatttctgg aacttttcta caaggaacga catagatcca ttaatgcgcc cctctaaaac
     7321 aatacccata atgtcttctt aagtatgcat ctgcttataa atgtttctgg atttacttta
     7381 gttagaatgt cgacacgaca atgaactatt gctgtagatc acgtgcgatc actgaattat
     7441 gagtttcttt ttatttttct ttttgggctt ttttttttgc aatttttttt ttccaccttt
     7501 gccaattgcg tagtcacata taatgtatac aatttttgca actctgacac tataattgat
     7561 ccaataataa agtatataat agatcatcaa gtatacgtaa tctccccaaa aaaaaaattg
     7621 cataaaataa aatctataaa ccttttttag atttttagtt gttccttttt tctttctatt
     7681 cttcaacttc caagtgttgt cactttcttc ctgttaacta tatttaaaag taaataccct
     7741 ccccagaaaa ctccggaatg agcctaggga cccgggggcg cgccggaggt ggaggttctg
     7801 gtggaggtgg ttcagctagc gatggtggtg ttcaattagc tgaccattat caacaaaata
     7861 ctccaattgg tgatggtcca gtcttgttac cagacaacca ttacttatcc tatcaatcta
     7921 aattatccaa agatccaaac gaaaagagag accacatggt cttgttagaa tttgttactg
     7981 ctgctggtat tacccatggt atggatgaat tgtacaaata aacgcgtagt acttaagatc
     8041 ctgaagacgg gatcgttcga agagtgaaat tctggaaatc tggaaatctg gttttgtatt
     8101 cttgttattc ttctttttgt tattacatat ataacttgtt acttttttaa aaaaatcttt
     8161 gtttatttta taaatatata aaactaaatt taagaaaaag agaaaaatgt tttatttgag
     8221 agattgaaat tttacttgaa tttagcttag cttttataaa gtattattat gtaaaaaaac
     8281 aaaacaaata tacattaaaa agttaagact ataaaatagc cacccaaggc atttctatat
     8341 cttgttgttg ttgttttcat cttctgtatc agaggaactt attttattat tttcgtcacg
     8401 ggtattttct cttgtttgat gattcatccc attcattcca tcataaaatt ctagaaggac
     8461 cacctttgat tgtaaatagt aataattacc acccttattt aattatttat ttaacttatt
     8521 tatttattta ttatacatat atacaaatct aataaagtga aaatctcccc cttcacccgg
     8581 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga
     8641 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga
     8701 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca
     8761 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc
     8821 cgaggagcag gactgacacg tgctaaaact tcatttttaa tttaaaagga tctaggtgaa
     8881 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     8941 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     9001 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     9061 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     9121 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     9181 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     9241 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     9301 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     9361 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     9421 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     9481 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     9541 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctgggctt
     9601 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     9661 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     9721 gtcagtgagc gaggaagcgg aag
//
