LOCUS       sequence-320746-        4889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFA-MNase-CaURA3 was double digested to remove the URA3 auxotrophy 
            marker. The SAT1 marker was amplified from pFA-SAT1 and cloned into 
            the AscI-SacI-digested pFA-MNase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4889)
  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 4889)
  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-SAT1
FEATURES             Location/Qualifiers
     source          1..4889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        85..103
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     85..102
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             164..229
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             236..682
                     /codon_start=1
                     /product="micrococcal nuclease from Staphylococcus aureus"
                     /label=MNase
                     /translation="ATSTKKLHKEPATLIKAIDGDTVKLMYKGQPMTFRLLLVDTPETK
                     HPKKGVEKYGPEASAFTKKMVENAKKIEVEFDKGQRTDKYGRGLAYIYADGKMVNEALV
                     RQGLAKVAYVYKPNNTHEQHLRKSEAQAKKEKLNIWSEDNADSGQ"
     primer_bind     complement(2627..2646)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2628..2646)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2733..2750)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2904..3492)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2984..3003)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3663..4454,4455..4523))
                     /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     4286..4305
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4524..4628)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4696..4714
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4752..4774)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     join(4874..4889,1..4)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgtactgaga gtgcaccata tggacatatt gtcgttagaa cgcggctaca attaatacat
       61 aaccttatgt atcatacaca tacgatttag gtgacactat agaacgcggc cgccagctga
      121 agcttcgtac gctgcaggtc gacggatccc cgggttaatt aaagattata aggaccatga
      181 cggagattac aaagatcacg acattgacta caaagatgac gacgataaag ctgccgccac
      241 atctaccaag aaacttcata aggaaccagc tacattgatt aaggcaatag atggggacac
      301 cgtaaaactt atgtacaaag ggcagccaat gaccttcaga ttgttgttgg ttgacactcc
      361 tgaaacaaaa cacccaaaga aaggagtgga aaaatatggt cctgaggctt ccgctttcac
      421 aaagaaaatg gtagaaaacg caaagaagat cgaagtcgag ttcgacaaag gtcaacgtac
      481 cgataaatat ggaagaggac ttgcatacat ttacgccgat ggaaaaatgg tgaatgaagc
      541 attggtgaga caagggttag caaaagttgc atatgtctac aagccaaata acacacacga
      601 gcagcattta cgtaaatcag aggcacaggc aaagaaagag aaattaaaca tttggagtga
      661 ggataatgct gattcagggc aatagtagag gcgcgccgtc gagcgtcaaa actagagaat
      721 aataaagaaa acgatctttt caaaaagaaa aaacctttta gttttccttt gttgttgttg
      781 tgggtgtgtg ctatttatat tatatagttt actcataata ccataaaata ttcggtttga
      841 ttaggttatt ttaataagct aatttgtttc taatcgtgta atttatgctg tgtatattaa
      901 gtagtgtgtg cactgcccaa aaatgtttgt tgtttatagt cggttaaaga gaaaaaagaa
      961 aaaaagatcc atacacacac gttaattagt tgttcaacgt aatacactca tattttgttc
     1021 ttatttgctt tcggtcgctg ttctcaccaa gatttattgc caacgaaaca attttttttt
     1081 atatattttc agatttttct ttttttcctt tcctttcctt ttctaatttt cactcctggt
     1141 tttctttctt tcttagaaac attatctcga tattaatatt aaaaaaatat aatcattcaa
     1201 aatggacggt ggtatgtttt agtttagctt caattctaat tgattgatta atcagttgat
     1261 tggtttcaat atgacaaatg ggtagggtgg gaaaacttca ttttcaattc agatcaaact
     1321 tttttgttgt cgacataata tttctcgttt gggatgttac tgtcacatta ataatacaca
     1381 cacatcagct tataattttg aaagtaattt atcagatatg ttgtgacgat cagtggaaat
     1441 ggctaacttc aatgtatctg ttcttcccct ttttcaaagt tcacgttttt tgattgattg
     1501 attgatctgt cggcagtggt ttcaaaacca ttcggtgagt aatcctatca atcaatgtta
     1561 cgacaaaagg ctcaatattc aaaattgcaa tgttttatgt tttcctacgt gtacttgtgc
     1621 aaggcaattg attcaacatt gcttttggtg tttgacgagt ttctagtttg gacttgtgtt
     1681 gttatctggg ctatacagat ttcccggctc actatgaatt ttttttttcg acgctcagtg
     1741 cacacaacta taaacaacac aaacacaaac acagcaagaa aaaaaaaaaa cgaacattga
     1801 attgaaacca agccaactga aaaattcctt atttaaatga ctgtcatact aacccatttt
     1861 tatagaagaa gttgctgctt tagttatcga taacggttct catatgaaaa tttcggtgat
     1921 ccctgagcag gtggcggaaa cattggatgc tgagaaccat ttcattgttc gtgaagtgtt
     1981 cgatgtgcac ctatccgacc aaggctttga actatctacc agaagtgtga gcccctaccg
     2041 gaaggattac atctcggatg atgactctga tgaagactct gcttgctatg gcgcattcat
     2101 cgaccaagag cttgtcggga agattgaact caactcaaca tggaacgatc tagcctctat
     2161 cgaacacatt gttgtgtcgc acacgcaccg aggcaaagga gtcgcgcaca gtctcatcga
     2221 atttgcgaaa aagtgggcac taagcagaca gctccttggc atacgattag agacacaaac
     2281 gaacaatgta cctgcctgca atttgtacgc aaaatgtggc tttactctcg gcggcattga
     2341 cctcttcacg tataaaacta gacctcaagt ctcgaacgaa acagcgatgt actggtactg
     2401 gttctcggga gcacaggatg acgcctaaca tatgtgaagt gtgaaggggg agattttcac
     2461 tttattagat ttgtatatat gtataataaa taaataaata agttaaataa ataattagat
     2521 aagggtggta attattacta tttacaatca aaggtggtcc tgagctcgaa ttcatcgatg
     2581 atatcagatc cactagtggc ctatgcggcc gcggatctgc cggtctccct atagtgagtc
     2641 gtattaattt cgataagcca ggttaacctg cattaatgaa tcggccaacg cgcggggaga
     2701 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     2761 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     2821 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2881 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2941 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3001 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3061 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3121 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3181 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3241 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3301 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3361 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3421 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3481 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3541 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3601 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3661 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3721 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3781 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3841 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3901 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3961 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4021 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4081 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4141 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4201 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4261 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4321 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4381 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4441 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4501 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4561 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4621 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     4681 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     4741 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     4801 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     4861 tggcttaact atgcggcatc agagcagat
//
