LOCUS       sequence-129162-        4694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ChEC-tagging vector encoding a short linker and aa 83-231 of Mnase 
            in pFA6a-3HA-kanMX6 replacing the 3xHA tag. These vectors retain 
            compatibility with the F2/R1 primer pairs commonly used for epitope.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4694)
  AUTHORS   Zentner GE, Kasinathan S, Xin B, Rohs R, Henikoff S
  TITLE     ChEC-seq kinetics discriminates transcription factor binding sites 
            by DNA sequence and shape in vivo.
  JOURNAL   Nat Commun. 2015 Oct 22;6:8733. doi: 10.1038/ncomms9733.
  PUBMED    26490019
REFERENCE   2  (bases 1 to 4694)
  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: "Nat 
            Commun."; date: "2015-10-22"; volume: "6:8733. doi"; pages: " 
            10.1038/ncomms9733"
COMMENT     Sequence Label: pGZ108 (pFA6a-3FLAG-MNase-kanMX6)
FEATURES             Location/Qualifiers
     source          1..4694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             63..128
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             135..581
                     /codon_start=1
                     /product="micrococcal nuclease from Staphylococcus aureus"
                     /label=MNase
                     /translation="ATSTKKLHKEPATLIKAIDGDTVKLMYKGQPMTFRLLLVDTPETK
                     HPKKGVEKYGPEASAFTKKMVENAKKIEVEFDKGQRTDKYGRGLAYIYADGKMVNEALV
                     RQGLAKVAYVYKPNNTHEQHLRKSEAQAKKEKLNIWSEDNADSGQ"
     terminator      609..796
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(664..688)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            871..2227
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        871..1214
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1215..2024
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1282..1301)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2030..2227
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2331..2350)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2332..2350)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2437..2454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2608..3196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2688..2707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3367..4158,4159..4227))
                     /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     3990..4009
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4228..4332)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4400..4418
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4456..4478)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4578..4597
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4678..4694,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4678..4694,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 aagattacaa ggatcacgat ggcgattaca aggatcacga tatcgattac aaggatgatg
      121 atgataaggc ggccgccacc agtaccaaga agctgcacaa ggagcccgcc accctgatca
      181 aggccataga tggcgatacc gtgaagctga tgtataaggg ccagcccatg accttccgcc
      241 tgctgctggt ggataccccc gagaccaagc acccgaaaaa gggcgtggaa aagtacggac
      301 ccgaggccag cgccttcaca aaaaagatgg tggagaacgc caagaaaatc gaggtggagt
      361 tcgataaagg ccaacgcacc gataagtatg gacgcggtct ggcctacatc tacgccgacg
      421 gcaaaatggt gaacgaggcc ctggtgcggc agggactggc caaggtggcg tacgtgtaca
      481 agcccaacaa cacccacgag cagcacctgc gcaagagcga ggctcaggca aagaaggaga
      541 aactgaacat ctggagtgag gataacgccg atagcggcca gtagtagagg cgcgccactt
      601 ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa
      661 taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga
      721 gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc gctcttattg
      781 accacacctc taccggcaga tccgctaggg ataacagggt aatatagatc tgtttagctt
      841 gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac
      901 agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata
      961 catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa
     1021 aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg
     1081 aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg
     1141 ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc
     1201 gaacataaac aaccatgggt aaggaaaaga ctcacgtttc gaggccgcga ttaaattcca
     1261 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     1321 cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     1381 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     1441 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     1501 ccactgcgat ccccggcaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     1561 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     1621 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     1681 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     1741 tctggaaaga aatgcataag cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     1801 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     1861 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     1921 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     1981 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagta ctgacaataa
     2041 aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct
     2101 attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga
     2161 tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc
     2221 tatactgctg tcgattcgat actaacgccg ccatccagtt taaacgagct cgaattcatc
     2281 gatgatatca gatccactag tggcctatgc ggccgcggat ctgccggtct ccctatagtg
     2341 agtcgtatta atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg
     2401 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2461 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2521 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2581 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2641 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2701 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2761 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     2821 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2881 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2941 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3001 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     3061 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3121 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3181 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3241 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3301 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3361 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3421 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3481 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3541 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3601 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3661 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3721 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3781 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3841 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3901 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3961 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4021 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4081 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4141 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4201 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4261 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4321 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4381 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4441 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4501 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4561 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatggacat
     4621 attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
     4681 taggtgacac tata
//
