LOCUS       sequence-181073-        5198 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of residues 1247-1451 from S. cerevisiae Spt6 
            with K1435A and K1355A mutations.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5198)
  AUTHORS   Sdano MA, Fulcher JM, Palani S, Chandrasekharan MB, Parnell TJ, 
            Whitby FG, Formosa T, Hill CP
  TITLE     A novel SH2 recognition mechanism recruits Spt6 to the doubly 
            phosphorylated RNA polymerase II linker at sites of transcription.
  JOURNAL   Elife. 2017 Aug 16;6. pii: 28723. doi: 10.7554/eLife.28723.
  PUBMED    28826505
REFERENCE   2  (bases 1 to 5198)
  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: "Elife."; 
            date: "2017-08-16"; volume: "6. pii: 28723. doi"; pages: " 
            10.7554/eLife.28723"
COMMENT     Sequence Label: CPH3506 (pET151_Spt6 1247-1451 K1355A,K1435A)
FEATURES             Location/Qualifiers
     source          1..5198
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     109..128
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     212..231
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        212..230
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    231..255
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             270..292
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             303..320
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             321..362
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             363..383
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1082..1100)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1096..1143
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1381..1399)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1467..1571
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1572..1640,1641..2432)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1790..1809)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2603..3191
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3092..3111
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3345..3362
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3377..3516
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3442..3461)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3602..3624
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3618..3809)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
       61 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
      121 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
      181 gtagaggatc gagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga
      241 gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga gatatacata
      301 tgcatcatca ccatcaccat ggtaagccta tccctaaccc tctcctcggt ctcgattcta
      361 cggaaaacct gtattttcag ggaattgatc ccttcaccgc aaagagaaca catcgtgtta
      421 tcaatcatcc ttactatttc cctttcaacg gcagacaggc tgaggattac ttaaggagta
      481 aagaacgtgg tgaattcgtg atcagacagt ctagccgagg tgatgaccac ttggttatca
      541 cctggaaatt ggataaggat ttgtttcaac atattgatat ccaagaatta gaaaaagaaa
      601 atcctttggc tttaggtaaa gtcttgattg tcgacaatca gaaatacaat gatttagacc
      661 agatcattgt agaatatctt caaaacaagg taaggctctt gaatgaaatg acatctagtg
      721 aagcgttcaa aagcggtact aagaaagatg tggtcaagtt tattgaagac tactctagag
      781 tgaatccaaa taagtctgtg tactatttca gtttgaacca cgataaccct ggttggtttt
      841 acttgatgtt caagattaac gcaaatagca aattatacac atggaatgtg aaattaacga
      901 acactggtta tttcctggta aactacaatt atccaagtgt tatccagctt tgtaatggtt
      961 ttgcgacgct tctaaaatct aacagtagta agaatagaat gaacaactac cgttagtaga
     1021 agggcgagct cagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     1081 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1141 ttgctgaaag gaggaactat atccggatat cccgcaagag gcccggcagt accggcataa
     1201 ccaagcctat gcctacagca tccagggtga cggtgccgag gatgacgatg agcgcattgt
     1261 tagatttcat acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt
     1321 aaagctagct tatcgatgat aagctgtcaa acatgagaat taattcttga agacgaaagg
     1381 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     1441 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1501 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1561 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1621 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1681 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1741 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1801 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1861 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1921 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1981 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     2041 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     2101 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     2161 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     2221 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     2281 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2341 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2401 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2461 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2521 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2581 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2641 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2701 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     2761 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2821 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2881 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2941 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3001 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3061 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3121 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3181 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3241 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3301 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3361 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3421 accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     3481 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     3541 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     3601 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     3661 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     3721 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     3781 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
     3841 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     3901 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     3961 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
     4021 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
     4081 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
     4141 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
     4201 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
     4261 atcatgcgca cccgtggcca ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg
     4321 gagatggcgg acgcgatgga tatgttctgc caagggttgg tttgcgcatt cacagttctc
     4381 cgcaagaatt gattggctcc aattcttgga gtggtgaatc cgttagcgag gtgccgccgg
     4441 cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga
     4501 caaggtatag ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg
     4561 cataaatcgc cgtgacgatc agcggtccaa tgatcgaagt taggctggta agagccgcga
     4621 gcgatccttg aagctgtccc tgatggtcgt catctacctg cctggacagc atggcctgca
     4681 acgcgggcat cccgatgccg ccggaagcga gaagaatcat aatggggaag gccatccagc
     4741 ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa
     4801 tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg
     4861 cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc
     4921 ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa
     4981 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
     5041 ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac
     5101 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5161 tgcattaatg aatcggccaa cgcgcgggga gaggcggt
//
