LOCUS       sequence-129170-        4619 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 4619)
  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 4619)
  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: pGZ173 (pFA6a-SL-MNase-kanMX6)
FEATURES             Location/Qualifiers
     source          1..4619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      534..721
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(589..613)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            796..2152
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        796..1139
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1140..1949
                     /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(1207..1226)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1955..2152
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2256..2275)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2257..2275)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2362..2379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2533..3121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2613..2632)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3292..4083,4084..4152))
                     /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     3915..3934
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4153..4257)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4325..4343
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4381..4403)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4503..4522
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4603..4619,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4603..4619,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatcg
       61 ccaccagtac caagaagctg cacaaggagc ccgccaccct gatcaaggcc atagatggcg
      121 ataccgtgaa gctgatgtat aagggccagc ccatgacctt ccgcctgctg ctggtggata
      181 cccccgagac caagcacccg aaaaagggcg tggaaaagta cggacccgag gccagcgcct
      241 ccacaaaaaa gatggtggag aacgccaaga aaatcgaggt ggagttcgat aaaggccaac
      301 gcaccgataa gtatggacgc ggtctggcct acatctacgc cgacggcaaa atggtgaacg
      361 aggccctggt gcggcaggga ctggccaagg tggcgtacgt gtacaagccc aacaacaccc
      421 acgagcagca cctgcgcaag agcgaggctc aggcaaagaa ggagaaactg aacatctgga
      481 gtgaggataa cgccgatagc ggccagtagt agaggcgcgc cacttctaaa taagcgaatt
      541 tcttatgatt tatgattttt attattaaat aagttataaa aaaaataagt gtatacaaat
      601 tttaaagtga ctcttaggtt ttaaaacgaa aattcttatt cttgagtaac tctttcctgt
      661 aggtcaggtt gctttctcag gtatagtatg aggtcgctct tattgaccac acctctaccg
      721 gcagatccgc tagggataac agggtaatat agatctgttt agcttgcctc gtccccgccg
      781 ggtcacccgg ccagcgacat ggaggcccag aataccctcc ttgacagtct tgacgtgcgc
      841 agctcagggg catgatgtga ctgtcgcccg tacatttagc ccatacatcc ccatgtataa
      901 tcatttgcat ccatacattt tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg
      961 ctgcagacct gcgagcaggg aaacgctccc ctcacagacg cgttgaattg tccccacgcc
     1021 gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt ctttcatata
     1081 cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca taaacaacca
     1141 tgggtaagga aaagactcac gtttcgaggc cgcgattaaa ttccaacatg gatgctgatt
     1201 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat
     1261 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     1321 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     1381 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     1441 gcaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     1501 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     1561 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     1621 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     1681 ataagctttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     1741 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     1801 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     1861 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     1921 ttcatttgat gctcgatgag tttttctaat cagtactgac aataaaaaga ttcttgtttt
     1981 caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt
     2041 tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc
     2101 gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat
     2161 tcgatactaa cgccgccatc cagtttaaac gagctcgaat tcatcgatga tatcagatcc
     2221 actagtggcc tatgcggccg cggatctgcc ggtctcccta tagtgagtcg tattaatttc
     2281 gataagccag gttaacctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2341 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2401 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2461 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2521 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2581 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2641 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2701 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     2761 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2821 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2881 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2941 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     3001 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3061 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3121 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3181 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3241 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3301 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3361 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3421 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3481 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3541 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3601 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3661 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3721 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3781 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3841 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3901 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3961 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4021 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4081 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4141 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4201 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4261 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     4321 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     4381 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     4441 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     4501 tgcggcatca gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg
     4561 cggctacaat taatacataa ccttatgtat catacacata cgatttaggt gacactata
//
