LOCUS       sequence-320745-        5231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DNA of the 3xFLAG epitope-MNase module for C. albicans. The 
            PacI-AscI 3xFLAG-MNase fragment was cloned in the PacI-AscI-digested
            pFA-TAP-CaARG4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5231)
  AUTHORS   Tebbji F, Khemiri I, Sellam A
  TITLE     High-Resolution Genome-Wide Occupancy in Candida spp. Using 
            ChEC-seq.
  JOURNAL   mSphere. 2020 Oct 14;5(5). pii: 5/5/e00646-20. doi: 
            10.1128/mSphere.00646-20.
  PUBMED    33055256
REFERENCE   2  (bases 1 to 5231)
  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: "mSphere."; 
            date: "2020-10-14"; volume: "5(5). pii: 5/5/e00646-20. doi"; pages: 
            " 10.1128/mSphere.00646-20"
COMMENT     Sequence Label: pFA-MNase-CaARG4
FEATURES             Location/Qualifiers
     source          1..5231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     16..35
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        116..134
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     116..133
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             195..260
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             267..713
                     /codon_start=1
                     /product="micrococcal nuclease from Staphylococcus aureus"
                     /label=MNase
                     /translation="ATSTKKLHKEPATLIKAIDGDTVKLMYKGQPMTFRLLLVDTPETK
                     HPKKGVEKYGPEASAFTKKMVENAKKIEVEFDKGQRTDKYGRGLAYIYADGKMVNEALV
                     RQGLAKVAYVYKPNNTHEQHLRKSEAQAKKEKLNIWSEDNADSGQ"
     primer_bind     complement(3000..3019)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3001..3019)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3106..3123)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3277..3865)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3357..3376)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4036..4827,4828..4896))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4659..4678
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4897..5001)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5069..5087
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5125..5147)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atggacatat
       61 tgtcgttaga acgcggctac aattaataca taaccttatg tatcatacac atacgattta
      121 ggtgacacta tagaacgcgg ccgccagctg aagcttcgta cgctgcaggt cgacggatcc
      181 ccgggttaat taaagattat aaggaccatg acggagatta caaagatcac gacattgact
      241 acaaagatga cgacgataaa gctgccgcca catctaccaa gaaacttcat aaggaaccag
      301 ctacattgat taaggcaata gatggggaca ccgtaaaact tatgtacaaa gggcagccaa
      361 tgaccttcag attgttgttg gttgacactc ctgaaacaaa acacccaaag aaaggagtgg
      421 aaaaatatgg tcctgaggct tccgctttca caaagaaaat ggtagaaaac gcaaagaaga
      481 tcgaagtcga gttcgacaaa ggtcaacgta ccgataaata tggaagagga cttgcataca
      541 tttacgccga tggaaaaatg gtgaatgaag cattggtgag acaagggtta gcaaaagttg
      601 catatgtcta caagccaaat aacacacacg agcagcattt acgtaaatca gaggcacagg
      661 caaagaaaga gaaattaaac atttggagtg aggataatgc tgattcaggg caatagtaga
      721 ggcgcgccat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
      781 ttatatcagt tattacccgg gaatctcggt cgtaatgatt tctataatga cgaaaaaaaa
      841 aaaattggaa agaaaaagct tcatggcctt tataaaaagg aactatccaa tacctcgcca
      901 gaaccaagta acagtatttt acggggcaca aatcaagaac aataagacag gactgtaaag
      961 atggacgcat agatcccccc tttagtaaga tttttcaaga gtagtctcaa ataaaccaga
     1021 atgaaggatt gaaggtttta ttcaataaat aattggacag aaagttattg tacagtttta
     1081 acaacaacaa tttcaaaatt tcaatttatg acatattgac cgacataatt cagtcatttc
     1141 taatattcaa ataacaagac accttgtata aatgaatcca caatcgtata tgaacaacca
     1201 aattttatga ctaactttgg gtcgtgtact tttgacgatt gagtcatttt taccaattag
     1261 gtaattgaat tttgttaatt tgtcattaac cgcgctggcc atcccaacaa taacaacaat
     1321 caatttaaaa acttgacaag gcaaaaaaaa aaaaatttta tctttgaaaa atttttcttc
     1381 cttttcttct tctgttcttt acataacagc tatcaagaat aatcaattaa taatgtcaca
     1441 acaacaagat aaacaaccta gtgaaaataa attatggggt ggtcgtttca ccggtgccac
     1501 tgatccattg atggatttat ataatgcttc attaccttat gataaagtta tgtatgatgc
     1561 tgatttaact ggtactaaag tatacacaca aggattaaac aaattaggat taattaccac
     1621 tgaagaattg catttgattc atcaaggatt agaacaaatc agacaagaat ggcatgataa
     1681 taaattcatt attaaggcag gtgatgagga tattcatact gctaatgaac gtcgattagg
     1741 agaaatcatt gggaaaaaca tttctggtaa agttcacact ggtagatcaa gaaatgatca
     1801 agttgctacc gatatgagaa ttttcgttcg tgaatcatta cttaatttaa gtaaaatttt
     1861 acatcaattc atcacggcaa ttcttgaacg ggcacataaa gaaatcgatg tattaatgcc
     1921 tggttatact catttacaaa aagctcaacc tattagatgg gcacattggt taagttcata
     1981 tgcaacttat ttcaccgaag attataaacg tttacaagaa attataactc gtgttaacca
     2041 atctccatta ggatctggtg ctttagcagg tcatccttat ggtattgatc gtgaattttt
     2101 agctaaagga ttaggatttg atggtgtcat tgggaattct ttaactgcag tttcagatcg
     2161 tgattttgtt gttgaaagtt tattttggag tacattattt atgaatcata tttctcgatt
     2221 ctcagaagat ctaattattt attctagtgg tgaatttggg ttcataaaat tagctgatgc
     2281 ttattctact ggttcttcat tgatgccaca aaagaaaaac cctgactcat tagaattatt
     2341 acgtgggaaa agtggtagag tatttggtca attatctgga tttttaatgt cgatcaaatc
     2401 tattccatct acttataata aagatatgca agaggataaa gaaccattat ttgatgcatt
     2461 gactacagtg gaacattcaa ttttaattgc tactggtgtg atttctactt tactgattga
     2521 taaacaaaat atggaaaaag ctttaactat ggatatgttg gctactgatt tagctgatta
     2581 tttagttaga aaaggagttc catttagaga aactcatcat atatctggtg aatgtgttag
     2641 aaaagctgaa gaagaaaaat tatctggaat tgatcaatta tcttttgaac aatttcaaca
     2701 aattgattct cgttttgaaa aagacgttat ggaaactttt gattttgaag ctagtgtgga
     2761 aagaagagat gctattggtg gtactgctaa aagtgccgtt ttaaaacaat tagaaaattt
     2821 aaaatcaatt ttaagttaat ttataaatag tcatataata atcacagtat tgtttaactt
     2881 acataaaata taatcaatta acacagagat accttgtact acaatagttt aaacgagctc
     2941 gaattcatcg atgatatcag atccactagt ggcctatgcg gccgcggatc tgccggtctc
     3001 cctatagtga gtcgtattaa tttcgataag ccaggttaac ctgcattaat gaatcggcca
     3061 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3121 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3181 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3241 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3301 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3361 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3421 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3481 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3541 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3601 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3661 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3721 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3781 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3841 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3901 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3961 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4021 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4081 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4141 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4201 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4261 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4321 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4381 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4441 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4501 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4561 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4621 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4681 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4741 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4801 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4861 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4921 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4981 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5041 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     5101 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     5161 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     5221 cgggtgtcgg g
//
