LOCUS       sequence-320701-        4623 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-CaURA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4623)
  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 4623)
  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-CaURA3
FEATURES             Location/Qualifiers
     source          1..4623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     12..31
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        112..130
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     112..129
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             191..256
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             263..709
                     /codon_start=1
                     /product="micrococcal nuclease from Staphylococcus aureus"
                     /label=MNase
                     /translation="ATSTKKLHKEPATLIKAIDGDTVKLMYKGQPMTFRLLLVDTPETK
                     HPKKGVEKYGPEASAFTKKMVENAKKIEVEFDKGQRTDKYGRGLAYIYADGKMVNEALV
                     RQGLAKVAYVYKPNNTHEQHLRKSEAQAKKEKLNIWSEDNADSGQ"
     primer_bind     complement(2388..2407)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2389..2407)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2494..2511)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2665..3253)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2745..2764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3424..4215,4216..4284))
                     /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     4047..4066
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4285..4389)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4457..4475
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4513..4535)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgg acatattgtc
       61 gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg
      121 acactataga acgcggccgc cagctgaagc ttcgtacgct gcaggtcgac ggatccccgg
      181 gttaattaaa gattataagg accatgacgg agattacaaa gatcacgaca ttgactacaa
      241 agatgacgac gataaagctg ccgccacatc taccaagaaa cttcataagg aaccagctac
      301 attgattaag gcaatagatg gggacaccgt aaaacttatg tacaaagggc agccaatgac
      361 cttcagattg ttgttggttg acactcctga aacaaaacac ccaaagaaag gagtggaaaa
      421 atatggtcct gaggcttccg ctttcacaaa gaaaatggta gaaaacgcaa agaagatcga
      481 agtcgagttc gacaaaggtc aacgtaccga taaatatgga agaggacttg catacattta
      541 cgccgatgga aaaatggtga atgaagcatt ggtgagacaa gggttagcaa aagttgcata
      601 tgtctacaag ccaaataaca cacacgagca gcatttacgt aaatcagagg cacaggcaaa
      661 gaaagagaaa ttaaacattt ggagtgagga taatgctgat tcagggcaat agtagaggcg
      721 cgccattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
      781 atcagttatt acccgggaat ctcggtcgta atgatttcta taatgacgaa aaaaaaaaaa
      841 ttggaaagaa aaagcttcat ggcctttata aaaaggaact atccaatacc tcgccagaac
      901 caagtaacag tattttacgg ggcacaaatc aagaacaata agacaggact gtaaagatgg
      961 acgcatagat ccggatggta taaacggaaa caaaaaaaag agctggtact actttcttta
     1021 aaattatttt attatttgat tttatttaat agtatatatt atattttgaa cgtagattat
     1081 tttgttgaaa gttgctgtag tgccattgat tcgtaacact aattctgtat tagtcattcc
     1141 tcttgtttga tagtatccaa aaaaacggct atttttttgc aatcttattt cctgcatatt
     1201 atacagataa cataatgaaa gaaaaaatct ttttttttgt tcttcaatga tgatttcaac
     1261 cattctttta aacattgatc aattcctgag caacaacccc atacacactg gtttatatat
     1321 cgcccctttt acagttgaag aaagaaatag aaatagaaat agcaaacaaa agatatgaca
     1381 gtcaacacta agacctatag tgagagagca gaaactcatg cctcaccagt agcacaacga
     1441 ttatttcgat taatggaact gaagaaaacc aatttatgtg catcaattga tgttgatacc
     1501 actaaggaat tccttgaatt aattgataaa ttgggtcctt atgtatgctt aatcaagact
     1561 catattgata taatcaatga tttttcctat gaatccacta ttgaaccatt attagaactt
     1621 tcacgtaaac atcaatttat gatttttgaa gatagaaaat ttgctgatat tggtaatacc
     1681 gtgaagaaac aatatattgg tggagtttat aaaattagta gttgggcaga tattactaat
     1741 gctcatggtg tcactgggaa tggagtagtt gaaggattaa aacagggagc taaagaaacc
     1801 accaccaacc aagagccaag agggttattg atgttagctg aattatcatc agtgggatca
     1861 ttagcatatg gagaatattc tcaaaaaact gttgaaattg ctaaatccga taaggaattt
     1921 gttgttggat ttattgccca acgtgatatg ggtggacaag aagaaggatt tgattggctt
     1981 attatgacac ctggagttgg attagatgat aaaggtgatg gattaggaca acaatataga
     2041 actgttgatg aagttgttag cactggaact gatattatca ttgttggtag aggattgttt
     2101 ggtaaaggaa gagatccaga tattgaaggt aaaaggtata gagatgctgg ttggaatgct
     2161 tatttgaaaa aaactggcca attataaatg tgaaggggga gattttcact ttattagatt
     2221 tgtatatatg tataataaat aaataaataa gttaaataaa taattaaata agggtggtaa
     2281 ttattactat ttacaatcaa aggtggtcct tctagtttaa acgagctcga attcatcgat
     2341 gatatcagat ccactagtgg cctatgcggc cgcggatctg ccggtctccc tatagtgagt
     2401 cgtattaatt tcgataagcc aggttaacct gcattaatga atcggccaac gcgcggggag
     2461 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     2521 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2581 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2641 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2701 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2761 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2821 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2881 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2941 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3001 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3061 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3121 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3181 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3241 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3301 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3361 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3421 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3481 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3541 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3601 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3661 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3721 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3781 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3841 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3901 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3961 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4021 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4081 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4141 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4201 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4261 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4321 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4381 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     4441 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     4501 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     4561 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     4621 ctg
//
