LOCUS       sequence-374282-        9461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone vector for generation of BiFC plasmids.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9461)
  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 9461)
  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: 2016-N
FEATURES             Location/Qualifiers
     source          1..9461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     42..59
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    176..197
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(212..217,218..235,236..242)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    250..266
                     /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     255..277
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     274..290
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     274..290
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        308..326
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     308..325
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4694..4996
                     /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     4875..4894
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             5345..6139
                     /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             5345..6139
                     /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(5399..5418)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6009..6028
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      6339..6348
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      8761..9349
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9250..9269
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
       61 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      121 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      181 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
      241 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
      301 ccaagctatt taggtgacac tatagaatac tcaagctatg catcaagctt ggtaccgagc
      361 tcggatccac tagtaacggc cgccagtgtg ctggaattcg cccttaagaa catttctgta
      421 ccgcaaatgt atcgactttt tcgatggtgt gttattgttg ttttgttgcg aaatataata
      481 gcttctgctt tatatccttt gtgttgatcc ttcccctcct ttcatacttg taaaattaca
      541 attggtattt tcaaccaata ttaccacacg tcatgcacta actcaaatgg gcaaaaaaaa
      601 gagagatagg aattacgtca ttttattatg tttttacgtg gactagaagg ttaagataat
      661 ttctggttat tttgttgata attgagtttt gttcgacctc caccaatctt taaactctcg
      721 ttgagtattc attttaatga ttttttgaaa tcagaagggg tgtttctaac ccaggattgt
      781 tttattgaga ggttgaaaca tcaatgtgtt agtttattga tttgggttgg tttaatagaa
      841 aaatatcagt aggtgttcaa taattgtagt ttattttctt ggctaaaatt gttgcgaaaa
      901 attgacaaaa taataacaaa aagtatttta gatgctaaaa cataatagta ggaatatcta
      961 cctgaaccat ttacattcaa caactggcaa gaaccttcct aataatttgc aagagtaatc
     1021 acaattaaac caaaatcaaa gtttaccgtg tatatagttg gacagaaagc tgtcacattg
     1081 gtttaataca tagttgaata tgtatgcatg aaacaacaac aatacaatta cagctcatga
     1141 tccttcaatt aatttcaaca ttcaaacaac ataacacctt gtgtaaatga ggttatgagt
     1201 acatgtgaac aacaaatctt atgactaaac tcgggtcgtg tacttttgac gattgagtca
     1261 tttttaccaa ttaggcaatt aatttttgct aaattttcat taaccgcgct ggccatccca
     1321 acaaacaata taaaagctga caacgaaaaa aaaaaaaana aaaactccaa ttctgaaaaa
     1381 tttttcttct tcttcatact aaacaaaaca actataaaga taatcaatta ataatgtcac
     1441 aacaacctag tgaaaataaa ttatggggtg gtcgattcac tggcgccact gatccattaa
     1501 tggatttata caatgcttca ttaccttatg ataaagttat gtatgatgca gatttaactg
     1561 gtactaaagt atatactcaa ggattgaaca aattaggatt aattaccact gaagaattga
     1621 atctgattca tcaaggttta gaacaaatta gacaagaatg gcatgataat aaatttatta
     1681 ttaaagctgg tgatgaagat attcatactg ctaatgaacg tagattagga gaaatcattg
     1741 ggaaaaacat ttctggtaaa gttcatactg gtagatccag aaatgatcaa gttgctactg
     1801 atatgagaat ttttgttcgt gaatcattac ttaatttaag taagatttta catcaattca
     1861 ttacagcaat tcttgaacga gctcataaag aaattgatgt attaatgcct ggttatactc
     1921 atttacaaaa agctcaacct attagatggg ctcattggtt aagttcatat gcaacttatt
     1981 tcactgaaga ttataaacgt ttacaagaaa tcattactcg tgttaatcaa tctccattag
     2041 gatctggtgc tttagctggt catccctacg gtattgatcg tgaattttta gctaaaggat
     2101 taggatttga tggtgttatt gggaattctt taactgcagt ttctgatcgt gattttgttg
     2161 ttgaaagttt attttggagt acattattta tgaatcatat ttctcgattt tcagaagatt
     2221 taattattta ttctagtggt gaatttggat ttataaagtt agctgatgct tactctactg
     2281 gttcatcatt aatgccacaa aagaaaaatc ctgattcatt agaattatta cgtgggaaaa
     2341 gtggcagagt gtttggtcag ttatctgggt ttttaatgtc aattaaatcc attccatcaa
     2401 cctataataa agatatgcaa gaagataaag aaccattatt tgatgcattg actaccgtag
     2461 aacattcaat tttaattgct actggtgtga tttctacttt actgattgat aaacaaaata
     2521 tggaaaaagc tttaactatg gatatgttag ctactgattt agcagattat ttagttagaa
     2581 aaggtgttcc atttagagaa actcatcata tttctggtga atgtgttaga aaagctgaag
     2641 aagaaaaatt gtcgggaatt gaccaattat catttgaaca atttcaacaa attgattcac
     2701 gttttgaaaa agatgttatg gaaacttttg attttgaagc tagtgtggaa agaagagatg
     2761 ctattggtgg tactgctaag agcgctgttt taaaacagtt agaaaatttg aaatcaattt
     2821 taaattaatt tataaataat catatagtat tcaatgtttt gttttgtttt tttttttttt
     2881 tttatggaaa tgtacaaaca gtatataatc aattaataga aagataccct gtattccatt
     2941 gtatcgcctt tgggatgtct ataataccaa ctttcaagaa tgagaaatta aacaacaatt
     3001 tctttcacca atcaactcgt ctatattcaa taaactcatt ctggtctttt caaatattta
     3061 atgttatcct gatcaaactt acaaatcaaa aatgtatcaa aatcattcat tagttcttct
     3121 tcttttgtta tctttgtcac tatatttctc tccatccaga ccaaaaatac ttccattgtg
     3181 aagatcgtct gaatcataat cagcaactac ctcttccatt tcagagaatt cagatttttc
     3241 aaaataggta tcacgggcca agtacacaca agacaaacga tacgagtgct gggatatttc
     3301 tttcaatgta tcactaaatt cgttgagatt taaagagagc cctttgactg ctttcattac
     3361 tttcctcaca tcatgagcat ccacattcaa atttcctgta gccctttatt cttagttttc
     3421 ttttcttatt cttttcttcg tacattgaaa tcacaaatgg tgatatcaaa cgaaggtcac
     3481 actgacttat gtctaatacc tccaatgatg atactggtaa cttaaccagg tctactcttt
     3541 tcaaacgatt tctcattagt gtcaatttac gtaggttctt caacctttca aacgatgcaa
     3601 atagcttttg taagggcgaa ttctgcagat atccatcaca ctggcggccg gccgctcgag
     3661 catgcatcta gagggccctt gaagaattgg aagaagttag tagttaaaaa aacaatggga
     3721 aagatttaat ctttttttct tttttcaaaa atttttaacc gctgatgata tattaatctc
     3781 tctccaccac tctctttaaa caccacaatt attaataata agatatctct cctactagtt
     3841 atttgttagt aagtatattg ttgattgatt ggttgggtta aaaatttttt ttttcataaa
     3901 gaaaataatg aaagtgagaa aaatttaaac cccacattac atactcaaca caacactaca
     3961 cacacgcatt taacaatcat catcagttga aatatatttt acaaacaatt acgattgttt
     4021 aatgaaaact atttctgttg ttattattat ttaacctttt ttttcttgtt gggttgaccc
     4081 tttttctatt aactatctaa ctaccacata acatttgaag ttgaactatt aaaccttaat
     4141 ttgcggccgc aaatctaatc tactttcttc ttcttctacc ctatttattc tgaaatgggg
     4201 gggggggggg gtggtgggaa ctttaggcgt gttaaatttg tcacatctgt tctcacttgc
     4261 ttctattcgt ataattttaa ttgtcttgtg agatcaatct tgttcttgtt cgagtttttt
     4321 cttgctttga ttaactacta ttattattag tatgcaatgc tgtttattac catttcataa
     4381 cttatattat aactcattga tttcaatacc tgctcaacaa tgagtcgtta ttattactgt
     4441 gctctgtatt ctgtactgct gcacgtacgt tatgtccatt gaagacaagc aaagaaagac
     4501 caacattcaa ttgtatctgc ttaccagagt tgtttacctc tgttaccaat atttggaaat
     4561 taattttttg gtggtggacg tttcttccta aaacacctag atgtgctagt ataattgcac
     4621 aaccagcacg cacactgcgt ctggccggtc ctggactaca ttttgtctct aacattcgta
     4681 gctgcacgta cggcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt
     4741 tgtggatgta cagagtgata ttattgacac gccggggcga cggatggtga tccccctggc
     4801 cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg
     4861 ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg
     4921 ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat
     4981 gttctgggga atataaatgt caggcatgag attatcaaaa aggatcttca cctagatcct
     5041 tttcacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg
     5101 ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct
     5161 tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc
     5221 tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc
     5281 gccaaggatc tgatggcgca ggggatcaag ctctgatcaa gagacaggat gaggatcgtt
     5341 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     5401 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     5461 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     5521 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     5581 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5641 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5701 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5761 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     5821 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     5881 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5941 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     6001 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     6061 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     6121 tcttgacgag ttcttctgaa ttattaacgc ttacaatttc ctgatgcggt attttctcct
     6181 tacgcatctg tgcggtattt cacaccgcat caggtggcac ttttcgggga aatgtgcgcg
     6241 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     6301 aaccctgata aatgcttcaa taatagcacg tgaggagggc caccatggcc aagttgacca
     6361 gtgccgttcc ggtgctcacc gcgcgcgacg tcgtctattc caagcctgtg gctttttttg
     6421 gtgattcaca atacataaat tggcaaatac ctcttctata gactttaacc tttgatgaaa
     6481 tggtatggat aaaccacttt tgtggtcagg gtatatatat ttttattcca atctggcctt
     6541 tataaattca aatttccaag gggactctgc taaaccttct gatataacta taactctaat
     6601 aatggctgta aaaagtgtga aaaaaagatg atattcacaa aaaagataaa ggggatttca
     6661 tattgggtat ctagatttaa acatcgtatt tcaacaatga aactagaatc aatacgaaat
     6721 ccagggcaca actgtgttca ttcccttggg aattatactg agtaatggga agaatgtgca
     6781 aaaaataccc gtgtcttata taatttccat ttgtgtaggg cttaaaagtg gacaacttca
     6841 ggttacgaaa ccacgaaaat gtctaaacgt ttgatcaaat tcgtaactca tggaaaatca
     6901 cagcaacaaa ttaaggatgg atatatccgg tacttacaaa taaaaacact aaagaagtat
     6961 aaattttgaa ctgaaattct atagaaaatc tttttttttg ggtaaaatta tgcgattgtg
     7021 gctcatagta acgtaatcat acagttcaac aaacatcatt ttgtcaaagg atttctaatc
     7081 ctattctttt tctacttggc ttttgagtat atatatatat atatatacat cctttgttca
     7141 tcatcatgtt cgattgaatt tttttcttgg ttgttattag attgtctact tgagtttttc
     7201 ttggtgcaaa ttgtggtgtt tttctaacaa atttttcaac atttaaagta tttctggaac
     7261 ttttctacaa ggaacgacat agatccatta atgcgcccct ctaaaacaat acccataatg
     7321 tcttcttaag tatgcatctg cttataaatg tttctggatt tactttagtt agaatgtcga
     7381 cacgacaatg aactattgct gtagatcacg tgcgatcact gaattatgag tttcttttta
     7441 tttttctttt tgggcttttt tttttgcaat tttttttttc cacctttgcc aattgcgtag
     7501 tcacatataa tgtatacaat ttttgcaact ctgacactat aattgatcca ataataaagt
     7561 atataataga tcatcaagta tacgtaatct ccccaaaaaa aaaattgcat aaaataaaat
     7621 ctataaacct tttttagatt tttagttgtt ccttttttct ttctattctt caacttccaa
     7681 gtgttgtcac tttcttcctg ttaactatat ttaaaagtaa ataccctccc cagaaaactc
     7741 cggagctagc ggaggtggag gttctggtgg aggtggttca agcctaggga cccgggggcg
     7801 cgcctaaacg cgtagtactt aagatcctga agacgggatc gttcgaagag tgaaattctg
     7861 gaaatctgga aatctggttt tgtattcttg ttattcttct ttttgttatt acatatataa
     7921 cttgttactt ttttaaaaaa atctttgttt attttataaa tatataaaac taaatttaag
     7981 aaaaagagaa aaatgtttta tttgagagat tgaaatttta cttgaattta gcttagcttt
     8041 tataaagtat tattatgtaa aaaaacaaaa caaatataca ttaaaaagtt aagactataa
     8101 aatagccacc caaggcattt ctatatcttg ttgttgttgt tttcatcttc tgtatcagag
     8161 gaacttattt tattattttc gtcacgggta ttttctcttg tttgatgatt catcccattc
     8221 attccatcat aaaattctag aaggaccacc tttgattgta aatagtaata attaccaccc
     8281 ttatttaatt atttatttaa cttatttatt tatttattat acatatatac aaatctaata
     8341 aagtgaaaat ctcccccttc acaccggtgt ggtccgggac gacgtgaccc tgttcatcag
     8401 cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct
     8461 ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg
     8521 gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc
     8581 ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc taaaacttca
     8641 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     8701 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     8761 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     8821 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     8881 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     8941 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     9001 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     9061 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     9121 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     9181 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     9241 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     9301 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     9361 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     9421 gttatcccct gattctgtgg ataaccgtat taccgccttt g
//
