LOCUS       sequence-377524-        8544 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 8544)
  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 8544)
  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: 2015-N
FEATURES             Location/Qualifiers
     source          1..8544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    117..138
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(153..158,159..176,177..183)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    191..207
                     /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     196..218
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     215..231
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     215..231
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        249..267
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     249..266
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2605..2624)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2606..2624)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2631..2648)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2631..2647)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2640..2662)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             3874..4176
                     /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     4055..4074
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             4525..5319
                     /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             4525..5319
                     /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(4579..4598)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5189..5208
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      5519..5528
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      7785..8373
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8274..8293
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8527..8544
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
       61 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
      121 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
      181 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
      241 gccaagctat ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag
      301 ctcggatcca ctagtaacgg ccgccagtgt gctggaattc gcccttggat ccaatcatca
      361 ctggtggggg tcaagaaaga ggtttgagat caggtacttt ggcaccacca ttagtagctg
      421 ggtttggtga agctgcaaga ttaatgaaac aagaatctgc atttgacaag aaacatattg
      481 agaggttatc tactaaattg aagaacggtt tgttatccat tccatctact cagtttaacg
      541 gttgtaataa cccaacatat caatacccag gttgtgtgaa tgtttccttt gcatacattg
      601 aaggggaatc tttgttaatg gcattgaaag atattgcatt gagttctggt tctgcatgta
      661 cctctgcatc tttggaacca tcgtatgttt tacatgcttt gggtgccgat gatgccttgg
      721 ctcattcttc cattagattc ggtattggta gattcaccac tgaagcagaa gttgactatg
      781 ttattcaagc aattaatgaa agagttgatt tcttgagaaa gatgtctcca ttatgggaaa
      841 tggttcaaga aggtattgac ttgaactcta tcgaatggag tggacattaa ggttttttgt
      901 ttttttgtaa tttgtgttgt tatgtgtaat tatcgtgtat agagtaattt cggaaattgt
      961 actttcgttc atgtatctga accagatgaa taatacgttc gtattgttga agaatgacta
     1021 actcctgtaa cggcgtgcgt gggttccgca gccgggttca attttttttt tcttgttcac
     1081 tataaaaata tataataaac gaaaaaaatc cgagggactc aggctatgaa atttttcact
     1141 caagaatttt ttttccttca tccttaattc ctttattcac tacccaaacc cacaatgtca
     1201 gttaaaacta aaactattac aattttacca ggtgatcacg tcggtaccga aattgtcaat
     1261 gaagctatca aagttttaga agcaattgaa gctgccactc cttaccaaaa aatccatttc
     1321 gatttcaaac atcatttaat cggtggtgct gccattgatg ccactggtgt cccattgcca
     1381 gatgacgcac ttgaaagtgc caaaaaatct gatgccgtct tattgggtgc tgttggtgga
     1441 ccaaaatggg ggactggcgc cgttcgtcca gaacaaggtt tattaaagat tcgtaaagaa
     1501 ttgaaccttt atgctaacat cagaccatgt aactttgcca gtgactcctt attggaatta
     1561 tccccattga gaccagaagt cgttaaaggt acaaacttga tcattgttcg tgaattagtt
     1621 ggtggtattt atttcggtga tcgtgaagaa caagaagaaa gtgaagataa acaaaccgca
     1681 tgggatactg aaaaatacac cgtcgatgaa gtcaccagaa tcacccgtat ggctgccttc
     1741 atggctttac aacataaccc accattacca atctggtcat tggataaggc taatgtttta
     1801 gcttcttcta gattatggag aagaactgtc gataaagtga tttctgaaga gttcccaact
     1861 ttgtctgttc aacatcaatt gattgactcc gccgccatga ttttaattca aaacccaacc
     1921 aaattgaatg gtataatcat cacttctaac atgtttggtg atatcatttc cgatgaagct
     1981 tcagttattc caggttcctt gggtttatta ccatctgcat ctttggcttc attgccagat
     2041 accaacaccg cttttggtct ttatgaacca tgtcacggtt ctgctcctga tttaccagcc
     2101 aataaagtta acccaattgc tactatttta tctgctgctt ctatgttgag attatctttg
     2161 gattgtgtga aagaagctga agctttagaa gaagccgtga aacaagtctt ggataaaggt
     2221 atcagaactg cagatttaag aggtaccagt tcagctactg aagttggtga cgcgattgtc
     2281 gaagctgtta ctaaaatctt aaaagaaaaa gcttaagtaa agtatacaca aaatttaacg
     2341 aatattatta atacaaaatg aacaccacct aaatactttt ttttaaaaat gtaccgactc
     2401 ttctaaaaat tgcgataatt caatatcgga aatttctctc accggtttac ttggatcttc
     2461 ggaaataaat ctatttgaag taatggtagc ctcacttgat tctgattggc tctttctaga
     2521 catgggtagg taagggcgaa ttctgcagat atccatcaca ctggcggccg gccgctcgag
     2581 catgcatcta gagggcccaa ttcgccctat agtgagtcgt attacaattc actggccgtc
     2641 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     2701 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg accagatcaa
     2761 aatgctagaa caaattacgt ctgcatcaac agtcatcttc cttatttgca taaatgctaa
     2821 aatggcaaac ctgtaaaccg tttgaaaagc aaattgagat agttagctcg gttttttttt
     2881 cacgtggcca taatccaaca aaaaaaagga atttaagggg ggagagaagg aggcttgaat
     2941 ttgtaccaca gagcgagcaa gaaattaata atatacttgg cttagctgaa atgagaatga
     3001 aaaagcaaat ttcaagtgaa acaccactac tagccaagta aattggccaa tttgctctcg
     3061 tgctttaatt tcctaacaag cttttgagac cgaaaaaaat caaccaatga tgatggaaaa
     3121 atacttccgg gagtcaattt atccataatt tcacgtcgtt ttttttttgc acatggcaaa
     3181 tatattggaa tcctgagatt ttcttggagt cgtgtacaat aacaccaggc gggaacatgc
     3241 cgttatgatc cagcatagcg gccgctatgt gttcaaatat tgaaaccgtg aaaccagttt
     3301 gatacagctg atattgtgtg ctaaaaaaag atgagcaccg aaagagtctt tagtttattg
     3361 ccaagtaaag ggtcaaagaa tacctattct ttatgaacac agacttgctc taatggtgaa
     3421 ttagtgtgga gtaatagaga aggaggcgaa acaaaaataa ttggttaaac actcgaatgt
     3481 cacacaatcc taaataggtt aaattcagtt accgccgtga tgcgggaaca aagccgagcg
     3541 gtaatatttt tggattctat gtttttttgg tgtatattgc acgacttggt gccaacaact
     3601 ggtgttccaa cgcacaaaac taggttcaac tggatactgt tttctagaaa ttatttgcgg
     3661 caattcatga ttacggtcct aaatttgaaa taaaagtttc caattatgtg tcttgacaat
     3721 ttccattggg ttcaaaaaga tctagaatgt atgtaattac gcaacaattt gcatgcaaag
     3781 aaataaaaac aaacctaaaa tccaactgct gttgttctgg tgatgatgta gtaaaataag
     3841 agcacggcca agaacaaggt gatttacgta cggcagttta aggtttacac ctataaaaga
     3901 gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac gccggggcga
     3961 cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt
     4021 tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt
     4081 gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc
     4141 aaaaacgcca ttaacctgat gttctgggga atataaatgt caggcatgag attatcaaaa
     4201 aggatcttca cctagatcct tttcacgtag aaagccagtc cgcagaaacg gtgctgaccc
     4261 cggatgaatg tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag
     4321 caggtagctt gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca
     4381 agcgaaccgg aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta
     4441 aactggatgg ctttcttgcc gccaaggatc tgatggcgca ggggatcaag ctctgatcaa
     4501 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4561 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4621 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4681 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     4741 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4801 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4861 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4921 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4981 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5041 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5101 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5161 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5221 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5281 cgcatcgcct tctatcgcct tcttgacgag ttcttctgaa ttattaacgc ttacaatttc
     5341 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat caggtggcac
     5401 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5461 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatagcacg tgaggagggc
     5521 caccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcttccaatt
     5581 agaggttgtg gatgcaaaat tgaaaaaaaa atgactattg gtagcgtgtt ctatttttct
     5641 tatgcgctga aaaactacga acaattgtct attccaagcc tgtggctttt tttggtgatt
     5701 cacaatacat aaattggcaa atacctcttc tatagacttt aacctttgat gaaatggtat
     5761 ggataaacca cttttgtggt cagggtatat atatttttat tccaatctgg cctttataaa
     5821 ttcaaatttc caaggggact ctgctaaacc ttctgatata actataactc taataatggc
     5881 tgtaaaaagt gtgaaaaaaa gatgatattc acaaaaaaga taaaggggat ttcatattgg
     5941 gtatctagat ttaaacatcg tatttcaaca atgaaactag aatcaatacg aaatccaggg
     6001 cacaactgtg ttcattccct tgggaattat actgagtaat gggaagaatg tgcaaaaaat
     6061 acccgtgtct tatataattt ccatttgtgt agggcttaaa agtggacaac ttcaggttac
     6121 gaaaccacga aaatgtctaa acgtttgatc aaattcgtaa ctcatggaaa atcacagcaa
     6181 caaattaagg atggatatat ccggtactta caaataaaaa cactaaagaa gtataaattt
     6241 tgaactgaaa ttctatagaa aatctttttt tttgggtaaa attatgcgat tgtggctcat
     6301 agtaacgtaa tcatacagtt caacaaacat cattttgtca aaggatttct aatcctattc
     6361 tttttctact tggcttttga gtatatatat atatatatat acatcctttg ttcatcatca
     6421 tgttcgattg aatttttttc ttggttgtta ttagattgtc tacttgagtt tttcttggtg
     6481 caaattgtgg tgtttttcta acaaattttt caacatttaa agtatttctg gaacttttct
     6541 acaaggaacg acatagatcc attaatgcgc ccctctaaaa caatacccat aatgtcttct
     6601 taagtatgca tctgcttata aatgtttctg gatttacttt agttagaatg tcgacacgac
     6661 aatgaactat tgctgtagat cacgtgcgat cactgaatta tgagtttctt tttatttttc
     6721 tttttgggct tttttttttg caattttttt tttccacctt tgccaattgc gtagtcacat
     6781 ataatgtata caatttttgc aactctgaca ctataattga tccaataata aagtatataa
     6841 tagatcatca agtatacgta atctccccaa aaaaaaaatt gcataaaata aaatctataa
     6901 acctttttta gatttttagt tgttcctttt ttctttctat tcttcaactt ccaagtgttg
     6961 tcactttctt cctgttaact atatttaaaa gtaaataccc tccccagaaa actccggagc
     7021 tagcggaggt ggaggttctg gtggaggtgg ttcaagccta gggacccggg ggcgcgccta
     7081 aacgcgtagt acttaaccgc gccgtcgacg agtgaaattc tggaaatctg gaaatctggt
     7141 tttgtattct tgttattctt ctttttgtta ttacatatat aacttgttac ttttttaaaa
     7201 aaatctttgt ttattttata aatatataaa actaaattta agaaaaagag aaaaatgttt
     7261 tatttgagag attgaaattt tacttgaatt tagcttagct tttataaagt attattatgt
     7321 aaaaaaacaa aacaaatata cattaaaaag ttaagactat aaaatagcca cccaaggcat
     7381 ttctatatct tgttgttgtt gttttcatct tctgtatcag aggaacttat tttattattt
     7441 tcgtcacggg tattttctct tgtttgatga ttcatcccat tcattccatc ataaaattct
     7501 agaaggacca cctttgattg taaatagtaa taattaccac ccttatttaa ttatttattt
     7561 aacttattta tttatttatt atacatatat acaaatctaa taaagtgaaa atctccccct
     7621 tcacacgtgg ccgaggagca ggactgacac gtgctaaaac ttcattttta atttaaaagg
     7681 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     7741 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     7801 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7861 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7921 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7981 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     8041 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     8101 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     8161 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     8221 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     8281 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     8341 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     8401 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     8461 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     8521 gagcgcagcg agtcagtgag cgag
//
