LOCUS       sequence-377526-        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-C
FEATURES             Location/Qualifiers
     source          1..8544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1017..1605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1506..1525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1759..1776
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1893..1914
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1929..1934,1935..1952,1953..1959)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1967..1983
                     /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     1972..1994
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1991..2007
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1991..2007
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2025..2043
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2025..2042
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4381..4400)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4382..4400)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4407..4424)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4407..4423)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4416..4438)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             5650..5952
                     /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     5831..5850
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             6301..7095
                     /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             6301..7095
                     /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(6355..6374)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6965..6984
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      7295..7304
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gcgtagtcac atataatgta tacaattttt gcaactctga cactataatt gatccaataa
       61 taaagtatat aatagatcat caagtatacg taatctcccc aaaaaaaaaa ttgcataaaa
      121 taaaatctat aaaccttttt tagattttta gttgttcctt ttttctttct attcttcaac
      181 ttccaagtgt tgtcactttc ttcctgttaa ctatatttaa aagtaaatac cctccccaga
      241 aaactccgga atgagcctag ggacccgggg gcgcgccgga ggtggaggtt ctggtggagg
      301 tggttcagct agcacgcgta gtacttaacc gcgccgtcga cgagtgaaat tctggaaatc
      361 tggaaatctg gttttgtatt cttgttattc ttctttttgt tattacatat ataacttgtt
      421 acttttttaa aaaaatcttt gtttatttta taaatatata aaactaaatt taagaaaaag
      481 agaaaaatgt tttatttgag agattgaaat tttacttgaa tttagcttag cttttataaa
      541 gtattattat gtaaaaaaac aaaacaaata tacattaaaa agttaagact ataaaatagc
      601 cacccaaggc atttctatat cttgttgttg ttgttttcat cttctgtatc agaggaactt
      661 attttattat tttcgtcacg ggtattttct cttgtttgat gattcatccc attcattcca
      721 tcataaaatt ctagaaggac cacctttgat tgtaaatagt aataattacc acccttattt
      781 aattatttat ttaacttatt tatttattta ttatacatat atacaaatct aataaagtga
      841 aaatctcccc cttcacacgt ggccgaggag caggactgac acgtgctaaa acttcatttt
      901 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
      961 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1021 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1081 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1141 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1201 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1261 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1321 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1381 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1441 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1501 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1561 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1621 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1681 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     1741 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     1801 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     1861 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     1921 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     1981 caatttcaca caggaaacag ctatgaccat gattacgcca agctatttag gtgacactat
     2041 agaatactca agctatgcat caagcttggt accgagctcg gatccactag taacggccgc
     2101 cagtgtgctg gaattcgccc ttggatccaa tcatcactgg tgggggtcaa gaaagaggtt
     2161 tgagatcagg tactttggca ccaccattag tagctgggtt tggtgaagct gcaagattaa
     2221 tgaaacaaga atctgcattt gacaagaaac atattgagag gttatctact aaattgaaga
     2281 acggtttgtt atccattcca tctactcagt ttaacggttg taataaccca acatatcaat
     2341 acccaggttg tgtgaatgtt tcctttgcat acattgaagg ggaatctttg ttaatggcat
     2401 tgaaagatat tgcattgagt tctggttctg catgtacctc tgcatctttg gaaccatcgt
     2461 atgttttaca tgctttgggt gccgatgatg ccttggctca ttcttccatt agattcggta
     2521 ttggtagatt caccactgaa gcagaagttg actatgttat tcaagcaatt aatgaaagag
     2581 ttgatttctt gagaaagatg tctccattat gggaaatggt tcaagaaggt attgacttga
     2641 actctatcga atggagtgga cattaaggtt ttttgttttt ttgtaatttg tgttgttatg
     2701 tgtaattatc gtgtatagag taatttcgga aattgtactt tcgttcatgt atctgaacca
     2761 gatgaataat acgttcgtat tgttgaagaa tgactaactc ctgtaacggc gtgcgtgggt
     2821 tccgcagccg ggttcaattt tttttttctt gttcactata aaaatatata ataaacgaaa
     2881 aaaatccgag ggactcaggc tatgaaattt ttcactcaag aatttttttt ccttcatcct
     2941 taattccttt attcactacc caaacccaca atgtcagtta aaactaaaac tattacaatt
     3001 ttaccaggtg atcacgtcgg taccgaaatt gtcaatgaag ctatcaaagt tttagaagca
     3061 attgaagctg ccactcctta ccaaaaaatc catttcgatt tcaaacatca tttaatcggt
     3121 ggtgctgcca ttgatgccac tggtgtccca ttgccagatg acgcacttga aagtgccaaa
     3181 aaatctgatg ccgtcttatt gggtgctgtt ggtggaccaa aatgggggac tggcgccgtt
     3241 cgtccagaac aaggtttatt aaagattcgt aaagaattga acctttatgc taacatcaga
     3301 ccatgtaact ttgccagtga ctccttattg gaattatccc cattgagacc agaagtcgtt
     3361 aaaggtacaa acttgatcat tgttcgtgaa ttagttggtg gtatttattt cggtgatcgt
     3421 gaagaacaag aagaaagtga agataaacaa accgcatggg atactgaaaa atacaccgtc
     3481 gatgaagtca ccagaatcac ccgtatggct gccttcatgg ctttacaaca taacccacca
     3541 ttaccaatct ggtcattgga taaggctaat gttttagctt cttctagatt atggagaaga
     3601 actgtcgata aagtgatttc tgaagagttc ccaactttgt ctgttcaaca tcaattgatt
     3661 gactccgccg ccatgatttt aattcaaaac ccaaccaaat tgaatggtat aatcatcact
     3721 tctaacatgt ttggtgatat catttccgat gaagcttcag ttattccagg ttccttgggt
     3781 ttattaccat ctgcatcttt ggcttcattg ccagatacca acaccgcttt tggtctttat
     3841 gaaccatgtc acggttctgc tcctgattta ccagccaata aagttaaccc aattgctact
     3901 attttatctg ctgcttctat gttgagatta tctttggatt gtgtgaaaga agctgaagct
     3961 ttagaagaag ccgtgaaaca agtcttggat aaaggtatca gaactgcaga tttaagaggt
     4021 accagttcag ctactgaagt tggtgacgcg attgtcgaag ctgttactaa aatcttaaaa
     4081 gaaaaagctt aagtaaagta tacacaaaat ttaacgaata ttattaatac aaaatgaaca
     4141 ccacctaaat actttttttt aaaaatgtac cgactcttct aaaaattgcg ataattcaat
     4201 atcggaaatt tctctcaccg gtttacttgg atcttcggaa ataaatctat ttgaagtaat
     4261 ggtagcctca cttgattctg attggctctt tctagacatg ggtaggtaag ggcgaattct
     4321 gcagatatcc atcacactgg cggccggccg ctcgagcatg catctagagg gcccaattcg
     4381 ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa
     4441 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     4501 aatagcgaag aggcccgcac cgatcgacca gatcaaaatg ctagaacaaa ttacgtctgc
     4561 atcaacagtc atcttcctta tttgcataaa tgctaaaatg gcaaacctgt aaaccgtttg
     4621 aaaagcaaat tgagatagtt agctcggttt ttttttcacg tggccataat ccaacaaaaa
     4681 aaaggaattt aaggggggag agaaggaggc ttgaatttgt accacagagc gagcaagaaa
     4741 ttaataatat acttggctta gctgaaatga gaatgaaaaa gcaaatttca agtgaaacac
     4801 cactactagc caagtaaatt ggccaatttg ctctcgtgct ttaatttcct aacaagcttt
     4861 tgagaccgaa aaaaatcaac caatgatgat ggaaaaatac ttccgggagt caatttatcc
     4921 ataatttcac gtcgtttttt ttttgcacat ggcaaatata ttggaatcct gagattttct
     4981 tggagtcgtg tacaataaca ccaggcggga acatgccgtt atgatccagc atagcggccg
     5041 ctatgtgttc aaatattgaa accgtgaaac cagtttgata cagctgatat tgtgtgctaa
     5101 aaaaagatga gcaccgaaag agtctttagt ttattgccaa gtaaagggtc aaagaatacc
     5161 tattctttat gaacacagac ttgctctaat ggtgaattag tgtggagtaa tagagaagga
     5221 ggcgaaacaa aaataattgg ttaaacactc gaatgtcaca caatcctaaa taggttaaat
     5281 tcagttaccg ccgtgatgcg ggaacaaagc cgagcggtaa tatttttgga ttctatgttt
     5341 ttttggtgta tattgcacga cttggtgcca acaactggtg ttccaacgca caaaactagg
     5401 ttcaactgga tactgttttc tagaaattat ttgcggcaat tcatgattac ggtcctaaat
     5461 ttgaaataaa agtttccaat tatgtgtctt gacaatttcc attgggttca aaaagatcta
     5521 gaatgtatgt aattacgcaa caatttgcat gcaaagaaat aaaaacaaac ctaaaatcca
     5581 actgctgttg ttctggtgat gatgtagtaa aataagagca cggccaagaa caaggtgatt
     5641 tacgtacggc agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg
     5701 gatgtacaga gtgatattat tgacacgccg gggcgacgga tggtgatccc cctggccagt
     5761 gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat
     5821 gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa
     5881 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc
     5941 tggggaatat aaatgtcagg catgagatta tcaaaaagga tcttcaccta gatccttttc
     6001 acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct
     6061 atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca
     6121 tggcgatagc tagactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg
     6181 gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca
     6241 aggatctgat ggcgcagggg atcaagctct gatcaagaga caggatgagg atcgtttcgc
     6301 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     6361 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     6421 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     6481 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     6541 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     6601 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     6661 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     6721 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     6781 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     6841 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6901 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6961 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     7021 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     7081 gacgagttct tctgaattat taacgcttac aatttcctga tgcggtattt tctccttacg
     7141 catctgtgcg gtatttcaca ccgcatcagg tggcactttt cggggaaatg tgcgcggaac
     7201 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     7261 ctgataaatg cttcaataat agcacgtgag gagggccacc atggccaagt tgaccagtgc
     7321 cgttccggtg ctcaccgcgc gcgacgtctt ccaattagag gttgtggatg caaaattgaa
     7381 aaaaaaatga ctattggtag cgtgttctat ttttcttatg cgctgaaaaa ctacgaacaa
     7441 ttgtctattc caagcctgtg gctttttttg gtgattcaca atacataaat tggcaaatac
     7501 ctcttctata gactttaacc tttgatgaaa tggtatggat aaaccacttt tgtggtcagg
     7561 gtatatatat ttttattcca atctggcctt tataaattca aatttccaag gggactctgc
     7621 taaaccttct gatataacta taactctaat aatggctgta aaaagtgtga aaaaaagatg
     7681 atattcacaa aaaagataaa ggggatttca tattgggtat ctagatttaa acatcgtatt
     7741 tcaacaatga aactagaatc aatacgaaat ccagggcaca actgtgttca ttcccttggg
     7801 aattatactg agtaatggga agaatgtgca aaaaataccc gtgtcttata taatttccat
     7861 ttgtgtaggg cttaaaagtg gacaacttca ggttacgaaa ccacgaaaat gtctaaacgt
     7921 ttgatcaaat tcgtaactca tggaaaatca cagcaacaaa ttaaggatgg atatatccgg
     7981 tacttacaaa taaaaacact aaagaagtat aaattttgaa ctgaaattct atagaaaatc
     8041 tttttttttg ggtaaaatta tgcgattgtg gctcatagta acgtaatcat acagttcaac
     8101 aaacatcatt ttgtcaaagg atttctaatc ctattctttt tctacttggc ttttgagtat
     8161 atatatatat atatatacat cctttgttca tcatcatgtt cgattgaatt tttttcttgg
     8221 ttgttattag attgtctact tgagtttttc ttggtgcaaa ttgtggtgtt tttctaacaa
     8281 atttttcaac atttaaagta tttctggaac ttttctacaa ggaacgacat agatccatta
     8341 atgcgcccct ctaaaacaat acccataatg tcttcttaag tatgcatctg cttataaatg
     8401 tttctggatt tactttagtt agaatgtcga cacgacaatg aactattgct gtagatcacg
     8461 tgcgatcact gaattatgag tttcttttta tttttctttt tgggcttttt tttttgcaat
     8521 tttttttttc cacctttgcc aatt
//
