LOCUS       sequence-377525-        9063 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BiFC plasmid for C-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 9063)
  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 9063)
  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: BiFC2
FEATURES             Location/Qualifiers
     source          1..9063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    93..114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(129..134,135..152,153..159)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    167..183
                     /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     172..194
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     191..207
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     191..207
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        225..243
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     225..242
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2581..2600)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2582..2600)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2607..2624)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2607..2623)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2616..2638)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             3850..4152
                     /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     4031..4050
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             4501..5295
                     /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             4501..5295
                     /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(4555..4574)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5165..5184
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      5495..5504
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      8280..8868
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8769..8788
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9022..9039
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
       61 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
      121 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
      181 caatttcaca caggaaacag ctatgaccat gattacgcca agctatttag gtgacactat
      241 agaatactca agctatgcat caagcttggt accgagctcg gatccactag taacggccgc
      301 cagtgtgctg gaattcgccc ttggatccaa tcatcactgg tgggggtcaa gaaagaggtt
      361 tgagatcagg tactttggca ccaccattag tagctgggtt tggtgaagct gcaagattaa
      421 tgaaacaaga atctgcattt gacaagaaac atattgagag gttatctact aaattgaaga
      481 acggtttgtt atccattcca tctactcagt ttaacggttg taataaccca acatatcaat
      541 acccaggttg tgtgaatgtt tcctttgcat acattgaagg ggaatctttg ttaatggcat
      601 tgaaagatat tgcattgagt tctggttctg catgtacctc tgcatctttg gaaccatcgt
      661 atgttttaca tgctttgggt gccgatgatg ccttggctca ttcttccatt agattcggta
      721 ttggtagatt caccactgaa gcagaagttg actatgttat tcaagcaatt aatgaaagag
      781 ttgatttctt gagaaagatg tctccattat gggaaatggt tcaagaaggt attgacttga
      841 actctatcga atggagtgga cattaaggtt ttttgttttt ttgtaatttg tgttgttatg
      901 tgtaattatc gtgtatagag taatttcgga aattgtactt tcgttcatgt atctgaacca
      961 gatgaataat acgttcgtat tgttgaagaa tgactaactc ctgtaacggc gtgcgtgggt
     1021 tccgcagccg ggttcaattt tttttttctt gttcactata aaaatatata ataaacgaaa
     1081 aaaatccgag ggactcaggc tatgaaattt ttcactcaag aatttttttt ccttcatcct
     1141 taattccttt attcactacc caaacccaca atgtcagtta aaactaaaac tattacaatt
     1201 ttaccaggtg atcacgtcgg taccgaaatt gtcaatgaag ctatcaaagt tttagaagca
     1261 attgaagctg ccactcctta ccaaaaaatc catttcgatt tcaaacatca tttaatcggt
     1321 ggtgctgcca ttgatgccac tggtgtccca ttgccagatg acgcacttga aagtgccaaa
     1381 aaatctgatg ccgtcttatt gggtgctgtt ggtggaccaa aatgggggac tggcgccgtt
     1441 cgtccagaac aaggtttatt aaagattcgt aaagaattga acctttatgc taacatcaga
     1501 ccatgtaact ttgccagtga ctccttattg gaattatccc cattgagacc agaagtcgtt
     1561 aaaggtacaa acttgatcat tgttcgtgaa ttagttggtg gtatttattt cggtgatcgt
     1621 gaagaacaag aagaaagtga agataaacaa accgcatggg atactgaaaa atacaccgtc
     1681 gatgaagtca ccagaatcac ccgtatggct gccttcatgg ctttacaaca taacccacca
     1741 ttaccaatct ggtcattgga taaggctaat gttttagctt cttctagatt atggagaaga
     1801 actgtcgata aagtgatttc tgaagagttc ccaactttgt ctgttcaaca tcaattgatt
     1861 gactccgccg ccatgatttt aattcaaaac ccaaccaaat tgaatggtat aatcatcact
     1921 tctaacatgt ttggtgatat catttccgat gaagcttcag ttattccagg ttccttgggt
     1981 ttattaccat ctgcatcttt ggcttcattg ccagatacca acaccgcttt tggtctttat
     2041 gaaccatgtc acggttctgc tcctgattta ccagccaata aagttaaccc aattgctact
     2101 attttatctg ctgcttctat gttgagatta tctttggatt gtgtgaaaga agctgaagct
     2161 ttagaagaag ccgtgaaaca agtcttggat aaaggtatca gaactgcaga tttaagaggt
     2221 accagttcag ctactgaagt tggtgacgcg attgtcgaag ctgttactaa aatcttaaaa
     2281 gaaaaagctt aagtaaagta tacacaaaat ttaacgaata ttattaatac aaaatgaaca
     2341 ccacctaaat actttttttt aaaaatgtac cgactcttct aaaaattgcg ataattcaat
     2401 atcggaaatt tctctcaccg gtttacttgg atcttcggaa ataaatctat ttgaagtaat
     2461 ggtagcctca cttgattctg attggctctt tctagacatg ggtaggtaag ggcgaattct
     2521 gcagatatcc atcacactgg cggccggccg ctcgagcatg catctagagg gcccaattcg
     2581 ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa
     2641 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     2701 aatagcgaag aggcccgcac cgatcgacca gatcaaaatg ctagaacaaa ttacgtctgc
     2761 atcaacagtc atcttcctta tttgcataaa tgctaaaatg gcaaacctgt aaaccgtttg
     2821 aaaagcaaat tgagatagtt agctcggttt ttttttcacg tggccataat ccaacaaaaa
     2881 aaaggaattt aaggggggag agaaggaggc ttgaatttgt accacagagc gagcaagaaa
     2941 ttaataatat acttggctta gctgaaatga gaatgaaaaa gcaaatttca agtgaaacac
     3001 cactactagc caagtaaatt ggccaatttg ctctcgtgct ttaatttcct aacaagcttt
     3061 tgagaccgaa aaaaatcaac caatgatgat ggaaaaatac ttccgggagt caatttatcc
     3121 ataatttcac gtcgtttttt ttttgcacat ggcaaatata ttggaatcct gagattttct
     3181 tggagtcgtg tacaataaca ccaggcggga acatgccgtt atgatccagc atagcggccg
     3241 ctatgtgttc aaatattgaa accgtgaaac cagtttgata cagctgatat tgtgtgctaa
     3301 aaaaagatga gcaccgaaag agtctttagt ttattgccaa gtaaagggtc aaagaatacc
     3361 tattctttat gaacacagac ttgctctaat ggtgaattag tgtggagtaa tagagaagga
     3421 ggcgaaacaa aaataattgg ttaaacactc gaatgtcaca caatcctaaa taggttaaat
     3481 tcagttaccg ccgtgatgcg ggaacaaagc cgagcggtaa tatttttgga ttctatgttt
     3541 ttttggtgta tattgcacga cttggtgcca acaactggtg ttccaacgca caaaactagg
     3601 ttcaactgga tactgttttc tagaaattat ttgcggcaat tcatgattac ggtcctaaat
     3661 ttgaaataaa agtttccaat tatgtgtctt gacaatttcc attgggttca aaaagatcta
     3721 gaatgtatgt aattacgcaa caatttgcat gcaaagaaat aaaaacaaac ctaaaatcca
     3781 actgctgttg ttctggtgat gatgtagtaa aataagagca cggccaagaa caaggtgatt
     3841 tacgtacggc agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg
     3901 gatgtacaga gtgatattat tgacacgccg gggcgacgga tggtgatccc cctggccagt
     3961 gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat
     4021 gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa
     4081 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc
     4141 tggggaatat aaatgtcagg catgagatta tcaaaaagga tcttcaccta gatccttttc
     4201 acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct
     4261 atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca
     4321 tggcgatagc tagactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg
     4381 gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca
     4441 aggatctgat ggcgcagggg atcaagctct gatcaagaga caggatgagg atcgtttcgc
     4501 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4561 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4621 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4681 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4741 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4801 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4861 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4921 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4981 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     5041 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5101 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5161 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5221 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5281 gacgagttct tctgaattat taacgcttac aatttcctga tgcggtattt tctccttacg
     5341 catctgtgcg gtatttcaca ccgcatcagg tggcactttt cggggaaatg tgcgcggaac
     5401 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5461 ctgataaatg cttcaataat agcacgtgag gagggccacc atggccaagt tgaccagtgc
     5521 cgttccggtg ctcaccgcgc gcgacgtctt ccaattagag gttgtggatg caaaattgaa
     5581 aaaaaaatga ctattggtag cgtgttctat ttttcttatg cgctgaaaaa ctacgaacaa
     5641 ttgtctattc caagcctgtg gctttttttg gtgattcaca atacataaat tggcaaatac
     5701 ctcttctata gactttaacc tttgatgaaa tggtatggat aaaccacttt tgtggtcagg
     5761 gtatatatat ttttattcca atctggcctt tataaattca aatttccaag gggactctgc
     5821 taaaccttct gatataacta taactctaat aatggctgta aaaagtgtga aaaaaagatg
     5881 atattcacaa aaaagataaa ggggatttca tattgggtat ctagatttaa acatcgtatt
     5941 tcaacaatga aactagaatc aatacgaaat ccagggcaca actgtgttca ttcccttggg
     6001 aattatactg agtaatggga agaatgtgca aaaaataccc gtgtcttata taatttccat
     6061 ttgtgtaggg cttaaaagtg gacaacttca ggttacgaaa ccacgaaaat gtctaaacgt
     6121 ttgatcaaat tcgtaactca tggaaaatca cagcaacaaa ttaaggatgg atatatccgg
     6181 tacttacaaa taaaaacact aaagaagtat aaattttgaa ctgaaattct atagaaaatc
     6241 tttttttttg ggtaaaatta tgcgattgtg gctcatagta acgtaatcat acagttcaac
     6301 aaacatcatt ttgtcaaagg atttctaatc ctattctttt tctacttggc ttttgagtat
     6361 atatatatat atatatacat cctttgttca tcatcatgtt cgattgaatt tttttcttgg
     6421 ttgttattag attgtctact tgagtttttc ttggtgcaaa ttgtggtgtt tttctaacaa
     6481 atttttcaac atttaaagta tttctggaac ttttctacaa ggaacgacat agatccatta
     6541 atgcgcccct ctaaaacaat acccataatg tcttcttaag tatgcatctg cttataaatg
     6601 tttctggatt tactttagtt agaatgtcga cacgacaatg aactattgct gtagatcacg
     6661 tgcgatcact gaattatgag tttcttttta tttttctttt tgggcttttt tttttgcaat
     6721 tttttttttc cacctttgcc aattgcgtag tcacatataa tgtatacaat ttttgcaact
     6781 ctgacactat aattgatcca ataataaagt atataataga tcatcaagta tacgtaatct
     6841 ccccaaaaaa aaaattgcat aaaataaaat ctataaacct tttttagatt tttagttgtt
     6901 ccttttttct ttctattctt caacttccaa gtgttgtcac tttcttcctg ttaactatat
     6961 ttaaaagtaa ataccctccc cagaaaactc cggaatgagc ctagggaccc gggggcgcgc
     7021 cggaggtgga ggttctggtg gaggtggttc agctagcatg tctaaaggtg aagaattatt
     7081 cactggtgtt gtcccaattt tggttgaatt agatggtgat gttaatggtc acaaattttc
     7141 tgtctccggt gaaggtgaag gtgatgctac ttacggtaaa ttgaccttaa aattgatttg
     7201 tactactggt aaattgccag ttccatggcc aaccttagtc actactttag gttatggttt
     7261 gcaatgtttt gctagatacc cagatcatat gaaacaacat gactttttca agtctgccat
     7321 gccagaaggt tatgttcaag aaagaactat ttttttcaaa gatgacggta actacaagac
     7381 cagagctgaa gtcaagtttg aaggtgatac cttagttaat agaatcgaat taaaaggtat
     7441 tgattttaaa gaagatggta acattttagg tcacaaattg gaatacaact ataactctca
     7501 caatgtttac atcactgctg acaaacaaaa gaatggtatc aaagctaact tcaaaattag
     7561 acacaacatt gaataaacgc gtagtactta accgcgccgt cgacgagtga aattctggaa
     7621 atctggaaat ctggttttgt attcttgtta ttcttctttt tgttattaca tatataactt
     7681 gttacttttt taaaaaaatc tttgtttatt ttataaatat ataaaactaa atttaagaaa
     7741 aagagaaaaa tgttttattt gagagattga aattttactt gaatttagct tagcttttat
     7801 aaagtattat tatgtaaaaa aacaaaacaa atatacatta aaaagttaag actataaaat
     7861 agccacccaa ggcatttcta tatcttgttg ttgttgtttt catcttctgt atcagaggaa
     7921 cttattttat tattttcgtc acgggtattt tctcttgttt gatgattcat cccattcatt
     7981 ccatcataaa attctagaag gaccaccttt gattgtaaat agtaataatt accaccctta
     8041 tttaattatt tatttaactt atttatttat ttattataca tatatacaaa tctaataaag
     8101 tgaaaatctc ccccttcaca cgtggccgag gagcaggact gacacgtgct aaaacttcat
     8161 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     8221 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     8281 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     8341 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     8401 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     8461 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     8521 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     8581 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     8641 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     8701 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     8761 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     8821 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     8881 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     8941 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     9001 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     9061 cgc
//
