LOCUS       sequence-129168-        4177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ChEC-tagging vector encoding a 3xFLAG tag and aa 83-231 of Mnase in 
            pFA6a-3HA-Trp1 replacing the 3xHA tag. These vectors retain 
            compatibility with the F2/R1 primer pairs commonly used for epitope 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4177)
  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 4177)
  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: pGZ110 (pFA6a-3FLAG-MNase-TRP1)
FEATURES             Location/Qualifiers
     source          1..4177
                     /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"
     promoter        832..979
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             980..1654
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     RBS             1734..1742
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(1814..1833)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1815..1833)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1920..1937)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2091..2679)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2171..2190)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2850..3641,3642..3710))
                     /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     3473..3492
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3711..3815)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3883..3901
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3939..3961)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4061..4080
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4161..4177,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4161..4177,1)
                     /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 tgtacaatct
      841 tgatccggag cttttctttt tttgccgatt aagaattaat tcggtcgaaa aaagaaaagg
      901 agagggccaa gagggagggc attggtgact attgagcacg tgagtatacg tgattaagca
      961 cacaaaggca gcttggagta tgtctgttat taatttcaca ggtagttctg gtccattggt
     1021 gaaagtttgc ggcttgcaga gcacagaggc cgcagaatgt gctctagatt ccgatgctga
     1081 cttgctgggt attatatgtg tgcccaatag aaagagaaca attgacccgg ttattgcaag
     1141 gaaaatttca agtcttgtaa aagcatataa aaatagttca ggcactccga aatacttggt
     1201 tggcgtgttt cgtaatcaac ctaaggagga tgttttggct ctggtcaatg attacggcat
     1261 tgatatcgtc caactgcatg gagatgagtc gtggcaagaa taccaagagt tcctcggttt
     1321 gccagttatt aaaagactcg tatttccaaa agactgcaac atactactca gtgcagcttc
     1381 acagaaacct cattcgttta ttcccttgtt tgattcagaa gcaggtggga caggtgaact
     1441 tttggattgg aactcgattt ctgactgggt tggaaggcaa gagagccccg aaagcttaca
     1501 ttttatgtta gctggtggac tgacgccaga aaatgttggt gatgcgctta gattaaatgg
     1561 cgttattggt gttgatgtaa gcggaggtgt ggagacaaat ggtgtaaaag actctaacaa
     1621 aatagcaaat ttcgtcaaaa atgctaagaa ataggttatt actgagtagt atttatttaa
     1681 gtattgtttg tgcacttgcc gatctatgcg gtgtgaaata ccgcacagat gcgtaaggag
     1741 gtttaaacga gctcgaattc atcgatgata tcagatccac tagtggccta tgcggccgcg
     1801 gatctgccgg tctccctata gtgagtcgta ttaatttcga taagccaggt taacctgcat
     1861 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     1921 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1981 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2041 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2101 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2161 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2221 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2281 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2341 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2401 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2461 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2521 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2581 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2641 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2701 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2761 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2821 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2881 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2941 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3001 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3061 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3121 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3181 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3241 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3301 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3361 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3421 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3481 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3541 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3601 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3661 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3721 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3781 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3841 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     3901 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     3961 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     4021 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     4081 ctgagagtgc accatatgga catattgtcg ttagaacgcg gctacaatta atacataacc
     4141 ttatgtatca tacacatacg atttaggtga cactata
//
