LOCUS       sequence-129164-        4538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ChEC-tagging vector encoding a 3xFLAG tag and aa 83-231 of Mnase in 
            pFA6a-3HA- HIS3MX6 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 4538)
  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 4538)
  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: pGZ109 (pFA6a-3FLAG-MNase-HIS3MX6)
FEATURES             Location/Qualifiers
     source          1..4538
                     /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..2071
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        871..1214
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1215..1868
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      1874..2071
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2175..2194)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2176..2194)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2281..2298)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2452..3040)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2532..2551)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3211..4002,4003..4071))
                     /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     3834..3853
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4072..4176)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4244..4262
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4300..4322)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4422..4441
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4522..4538,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4522..4538,1)
                     /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 aggagggctt ttgtagaaag aaatacgaac gaaacgaaaa
     1261 tcagcgttgc catcgctttg gacaaagctc ccttacctga agagtcgaat tttattgatg
     1321 aacttataac ttccaagcat gcaaaccaaa agggagaaca agtaatccaa gtagacacgg
     1381 gaattggatt cttggatcac atgtatcatg cactggctaa acatgcaggc tggagcttac
     1441 gactttactc aagaggtgat ttaatcatcg atgatcatca cactgcagaa gatactgcta
     1501 ttgcacttgg tattgcattc aagcaggcta tgggtaactt tgccggcgtt aaaagatttg
     1561 gacatgctta ttgtccactt gacgaagctc tttctagaag cgtagttgac ttgtcgggac
     1621 ggccctatgc tgttatcgat ttgggattaa agcgtgaaaa ggttggggaa ttgtcctgtg
     1681 aaatgatccc tcacttacta tattcctttt cggtagcagc tggaattact ttgcatgtta
     1741 cctgcttata tggtagtaat gaccatcatc gtgctgaaag cgcttttaaa tctctggctg
     1801 ttgccatgcg cgcggctact agtcttactg gaagttctga agtcccaagc acgaagggag
     1861 tgttgtaaag gatactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata
     1921 gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt
     1981 tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt
     2041 caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac gccgccatcc
     2101 agtttaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct atgcggccgc
     2161 ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca
     2221 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2281 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2341 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2401 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2461 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2521 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2581 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2641 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2701 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2761 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2821 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2881 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2941 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3001 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3061 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3121 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3181 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3241 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3301 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3361 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3421 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3481 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3541 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3601 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3661 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3721 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3781 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3841 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3901 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3961 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4021 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4081 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4141 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4201 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     4261 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     4321 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     4381 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     4441 actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac
     4501 cttatgtatc atacacatac gatttaggtg acactata
//
