LOCUS       sequence-106351-        9774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  prey plasmid for an adapted two-hybrid system for C. albicans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9774)
  AUTHORS   Stynen B, Van Dijck P, Tournu H
  TITLE     A CUG codon adapted two-hybrid system for the pathogenic fungus 
            Candida albicans.
  JOURNAL   Nucleic Acids Res. 2010 Oct;38(19):e184. doi: 10.1093/nar/gkq725. 
            Epub 2010 Aug 17.
  PUBMED    20719741
REFERENCE   2  (bases 1 to 9774)
  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"; journalName: "Nucleic 
            Acids Res."; date: "2010-10"; volume: "38(19):e184. doi"; pages: " 
            10.1093/nar/gkq725. Epub 2010 Aug 17"
COMMENT     Sequence Label: pC2HP
FEATURES             Location/Qualifiers
     source          1..9774
                     /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             4624..4926
                     /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     4805..4824
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             5275..6069
                     /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             5275..6069
                     /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(5329..5348)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5939..5958
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      6269..6278
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7776..7796
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7947..8012
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     rep_origin      9005..9593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9494..9513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9747..9764
                     /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 ctagtaacgg ccgccagtgt gctggaattc gcccttaaga acatttctgt accgcaaatg
      361 tatcgacttt ttcgatggtg tgttattgtt gttttgttgc gaaatataat agcttctgct
      421 ttatatcctt tgtgttgatc cttcccctcc tttcatactt gtaaaattac aattggtatt
      481 ttcaaccaat attaccacac gtcatgcact aactcaaatg ggcaaaaaaa agagagatag
      541 gaattacgtc attttattat gtttttacgt ggactagaag gttaagataa tttctggtta
      601 ttttgttgat aattgagttt tgttcgacct ccaccaatct ttaaactctc gttgagtatt
      661 cattttaatg attttttgaa atcagaaggg gtgtttctaa cccaggattg ttttattgag
      721 aggttgaaac atcaatgtgt tagtttattg atttgggttg gtttaataga aaaatatcag
      781 taggtgttca ataattgtag tttattttct tggctaaaat tgttgcgaaa aattgacaaa
      841 ataataacaa aaagtatttt agatgctaaa acataatagt aggaatatct acctgaacca
      901 tttacattca acaactggca agaaccttcc taataatttg caagagtaat cacaattaaa
      961 ccaaaatcaa agtttaccgt gtatatagtt ggacagaaag ctgtcacatt ggtttaatac
     1021 atagttgaat atgtatgcat gaaacaacaa caatacaatt acagctcatg atccttcaat
     1081 taatttcaac attcaaacaa cataacacct tgtgtaaatg aggttatgag tacatgtgaa
     1141 caacaaatct tatgactaaa ctcgggtcgt gtacttttga cgattgagtc atttttacca
     1201 attaggcaat taatttttgc taaattttca ttaaccgcgc tggccatccc aacaaacaat
     1261 ataaaagctg acaacgaaaa aaaaaaaaaa aaaaactcca attctgaaaa atttttcttc
     1321 ttcttcatac taaacaaaac aactataaag ataatcaatt aataatgtca caacaaccta
     1381 gtgaaaataa attatggggt ggtcgattca ctggcgccac tgatccatta atggatttat
     1441 acaatgcttc attaccttat gataaagtta tgtatgatgc agatttaact ggtactaaag
     1501 tatatactca aggattgaac aaattaggat taattaccac tgaagaattg aatctgattc
     1561 atcaaggttt agaacaaatt agacaagaat ggcatgataa taaatttatt attaaagctg
     1621 gtgatgaaga tattcatact gctaatgaac gtagattagg agaaatcatt gggaaaaaca
     1681 tttctggtaa agttcatact ggtagatcca gaaatgatca agttgctact gatatgagaa
     1741 tttttgttcg tgaatcatta cttaatttaa gtaagatttt acatcaattc attacagcaa
     1801 ttcttgaacg agctcataaa gaaattgatg tattaatgcc tggttatact catttacaaa
     1861 aagctcaacc tattagatgg gctcattggt taagttcata tgcaacttat ttcactgaag
     1921 attataaacg tttacaagaa atcattactc gtgttaatca atctccatta ggatctggtg
     1981 ctttagctgg tcatccctac ggtattgatc gtgaattttt agctaaagga ttaggatttg
     2041 atggtgttat tgggaattct ttaactgcag tttctgatcg tgattttgtt gttgaaagtt
     2101 tattttggag tacattattt atgaatcata tttctcgatt ttcagaagat ttaattattt
     2161 attctagtgg tgaatttgga tttataaagt tagctgatgc ttactctact ggttcatcat
     2221 taatgccaca aaagaaaaat cctgattcat tagaattatt acgtgggaaa agtggcagag
     2281 tgtttggtca gttatctggg tttttaatgt caattaaatc cattccatca acctataata
     2341 aagatatgca agaagataaa gaaccattat ttgatgcatt gactaccgta gaacattcaa
     2401 ttttaattgc tactggtgtg atttctactt tactgattga taaacaaaat atggaaaaag
     2461 ctttaactat ggatatgtta gctactgatt tagcagatta tttagttaga aaaggtgttc
     2521 catttagaga aactcatcat atttctggtg aatgtgttag aaaagctgaa gaagaaaaat
     2581 tgtcgggaat tgaccaatta tcatttgaac aatttcaaca aattgattca cgttttgaaa
     2641 aagatgttat ggaaactttt gattttgaag ctagtgtgga aagaagagat gctattggtg
     2701 gtactgctaa gagcgctgtt ttaaaacagt tagaaaattt gaaatcaatt ttaaattaat
     2761 ttataaataa tcatatagta ttcaatgttt tgttttgttt tttttttttt tttatggaaa
     2821 tgtacaaaca gtatataatc aattaataga aagataccct gtattccatt gtatcgcctt
     2881 tgggatgtct ataataccaa ctttcaagaa tgagaaatta aacaacaatt tctttcacca
     2941 atcaactcgt ctatattcaa taaactcatt ctggtctttt caaatattta atgttatcct
     3001 gatcaaactt acaaatcaaa aatgtatcaa aatcattcat tagttcttct tcttttgtta
     3061 tctttgtcac tatatttctc tccatccaga ccaaaaatac ttccattgtg aagatcgtct
     3121 gaatcataat cagcaactac ctcttccatt tcagagaatt cagatttttc aaaataggta
     3181 tcacgggcca agtacacaca agacaaacga tacgagtgct gggatatttc tttcaatgta
     3241 tcactaaatt cgttgagatt taaagagagc cctttgactg ctttcattac tttcctcaca
     3301 tcatgagcat ccacattcaa atttcctgta gccctttatt cttagttttc ttttcttatt
     3361 cttttcttcg tacattgaaa tcacaaatgg tgatatcaaa cgaaggtcac actgacttat
     3421 gtctaatacc tccaatgatg atactggtaa cttaaccagg tctactcttt tcaaacgatt
     3481 tctcattagt gtcaatttac gtaggttctt caacctttca aacgatgcaa atagcttttg
     3541 taagggcgaa ttctgcagat atccatcaca ctggcggccg gccgctcgag catgcatcta
     3601 gagggccctt gaagaattgg aagaagttag tagttaaaaa aacaatggga aagatttaat
     3661 ctttttttct tttttcaaaa atttttaacc gctgatgata tattaatctc tctccaccac
     3721 tctctttaaa caccacaatt attaataata agatatctct cctactagtt atttgttagt
     3781 aagtatattg ttgattgatt ggttgggtta aaaatttttt ttttcataaa gaaaataatg
     3841 aaagtgggaa aaatttaaac cccacattac atactcaaca caacactaca cacacgcatt
     3901 taacaatcat catcagttga aatatatttt acaaacaatt acgattgttt aatgaaaact
     3961 atttctgttg ttattattat ttaacctttt ttttcttgtt gggttgaccc tttttctatt
     4021 aactatctaa ctaccacata acatttgaag ttgaactatt aaaccttaat ttgcggccgc
     4081 aaatctaatc tactttcttc ttcttctacc ctatttattc tgaaatgggg gggggggggg
     4141 gtggtgggaa ctttaggcgt gttaaatttg tcacatctgt tctcacttgc ttctattcgt
     4201 ataattttaa ttgtcttgtg agatcaatct tgttcttgtt cgagtttttt cttgctttga
     4261 ttaactacta ttattattag tatgcaatgc tgtttattac catttcataa cttatattat
     4321 aactcattga tttcaatacc tgctcaacaa tgagtcgtta ttattactgt gctctgtatt
     4381 ctgtactgct gcacgtacgt tatgtccatt gaagacaagc aaagaaagac caacattcaa
     4441 ttgtatctgc ttaccagagt tgtttacctc tgttaccaat atttggaaat taattttttg
     4501 gtggtggacg tttcttccta aaacacctag atgtgctagt ataattgcac aaccagcacg
     4561 cacactgcgt ctggccggtc ctggactaca ttttgtctct aacattcgta gctgcacgta
     4621 cggcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
     4681 cagagtgata ttattgacac gccggggcga cggatggtga tccccctggc cagtgcacgt
     4741 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
     4801 tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg
     4861 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga
     4921 atataaatgt caggcatgag attatcaaaa aggatcttca cctagatcct tttcacgtag
     4981 aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg
     5041 acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga
     5101 tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc
     5161 tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc
     5221 tgatggcgca ggggatcaag ctctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5281 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5341 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5401 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     5461 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     5521 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     5581 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     5641 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     5701 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5761 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     5821 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5881 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5941 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6001 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6061 ttcttctgaa ttattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
     6121 tgcggtattt cacaccgcat caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6181 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6241 aatgcttcaa taatagcacg tgaggagggc caccatggcc aagttgacca gtgccgttcc
     6301 ggtgctcacc gcgcgcgacg tcgtctattc caagcctgtg gctttttttg gtgattcaca
     6361 atacataaat tggcaaatac ctcttctata gactttaacc tttgatgaaa tggtatgcta
     6421 ttccaagcct gtggcttttt ttggtgattc acaatacata aattggcaaa tacctcttct
     6481 atagacttta acctttgatg aaatggtatg gataaaccac ttttgtggtc agggtatata
     6541 tatttttatt ccaatctggc ctttataaat tcaaatttcc aaggggactc tgctaaacct
     6601 tctgatataa ctataactct aataatggct gtaaaaagtg tgaaaaaaag atgatattca
     6661 caaaaaagat aaaggggatt tcatattggg tatctagatt taaacatcgt atttcaacaa
     6721 tgaaactaga atcaatacga aatccagggc acaactgtgt tcattccctt gggaattata
     6781 ctgagtaatg ggaagaatgt gcaaaaaata cccgtgtctt atataatttc catttgtgta
     6841 gggcttaaaa gtggacaact tcaggttacg aaaccacgaa aatgtctaaa cgtttgatca
     6901 aattcgtaac tcatggaaaa tcacagcaac aaattaagga tggatatatc cggtacttac
     6961 aaataaaaac actaaagaag tataaatttt gaactgaaat tctatagaaa atcttttttt
     7021 ttgggtaaaa ttatgcgatt gtggctcata gtaacgtaat catacagttc aacaaacatc
     7081 attttgtcaa aggatttcta atcctattct ttttctactt ggcttttgag tatatatata
     7141 tatatatata catcctttgt tcatcatcat gttcgattga atttttttct tggttgttat
     7201 tagattgtct acttgagttt ttcttggtgc aattgtggtg ttttctaaca atttttcaac
     7261 attaaagtat ttctggaact tttctacaag gaacgacata gatccattaa tgcgcccctc
     7321 taaaacaata cccataatgt cttcttaagt atgcatctgc ttataaatgt ttctggattt
     7381 actttagtta gaatgtcgac acgacaatga actattgctg tagatcacgt gcgatcactg
     7441 aattatgagt ttctttttat ttttcttttt gggctttttt ttttgcaatt ttttttttcc
     7501 acctttgcca attgcgtagt cacatataat gtatacaatt tttgcaactc tgacactata
     7561 attgatccaa taataaagta tataatagat catcaagtat acgtaatctc cccaaaaaaa
     7621 aaattgcata aaataaaatc tataaacctt ttttagattt ttagttgttc cttttttctt
     7681 tctattcttc aacttccaag tgttgtcact ttcttcctgt taactatatt taaaagtaaa
     7741 taccctcccc agaaaactcc ggaatgggtg ctcctccaaa aaagaagaga aaggtagctc
     7801 cgcggtctcc agggcccggt ttcaccccac acgattctgc tccatacggt gctttggata
     7861 tggctgattt cgaattcgaa caaatgttca ccgatgcttt gggtattgat gaatacggtg
     7921 gtgctagcgg tgctggtggt gttggtgact acaaagacca tgacggtgat tataaagatc
     7981 atgacatcga ttacaaggat gacgatgaca agggagcagg aggcgtcgga aggcctcccg
     8041 ggaggcgcgc cacgcgtagt acttaagatc ctgaagacgg gatcgttcga agagtgaaat
     8101 tctggaaatc tggaaatctg gttttgtatt cttgttattc ttctttttgt tattacatat
     8161 ataacttgtt acttttttaa aaaaatcttt gtttatttta taaatatata aaactaaatt
     8221 taagaaaaag agaaaaatgt tttatttgag agattgaaat tttacttgaa tttagcttag
     8281 cttttataaa gtattattat gtaaaaaaac aaaacaaata tacattaaaa agttaagact
     8341 ataaaatagc cacccaaggc atttctatat cttgttgttg ttgttttcat cttctgtatc
     8401 agaggaactt attttattat tttcgtcacg ggtattttct cttgtttgat gattcatccc
     8461 attcattcca tcataaaatt ctagaaggac cacctttgat tgtaaatagt aataattacc
     8521 acccttattt aattatttat ttaacttatt tatttattta ttatacatat atacaaatct
     8581 aataaagtga aaatctcccc cttcacaccg gtgtggtccg ggacgacgtg accctgttca
     8641 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     8701 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     8761 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     8821 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctaaaac
     8881 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     8941 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     9001 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     9061 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     9121 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     9181 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     9241 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     9301 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     9361 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcctcccg
     9421 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     9481 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     9541 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     9601 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     9661 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     9721 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaag
//
