LOCUS       sequence-181069-        6378 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of residues 1247-1451 from S. cerevisiae Spt6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6378)
  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 6378)
  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: CPH1141 (pET151_Spt6 1247-1451)
FEATURES             Location/Qualifiers
     source          1..6378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     108..127
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     211..230
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        211..229
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    230..254
                     /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             269..291
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             302..319
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             320..361
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             362..382
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1081..1099)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1095..1142
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1380..1398)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1466..1570
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1571..1639,1640..2431)
                     /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(1789..1808)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2602..3190
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3091..3110
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3344..3361
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3376..3515
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3441..3460)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3601..3623
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3617..3808)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5120..6202)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6164..6183
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6203..6280)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
       61 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
      121 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
      181 tagaggatcg agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag
      241 cggataacaa ttcccctcta gaaataattt tgtttaactt taagaaggag atatacatat
      301 gcatcatcac catcaccatg gtaagcctat ccctaaccct ctcctcggtc tcgattctac
      361 ggaaaacctg tattttcagg gaattgatcc cttcaccgca aagagaacac atcgtgttat
      421 caatcatcct tactatttcc ctttcaacgg cagacaggct gaggattact taaggagtaa
      481 agaacgtggt gaattcgtga tcagacagtc tagccgaggt gatgaccact tggttatcac
      541 ctggaaattg gataaggatt tgtttcaaca tattgatatc caagaattag aaaaagaaaa
      601 tcctttggct ttaggtaaag tcttgattgt cgacaatcag aaatacaatg atttagacca
      661 gatcattgta gaatatcttc aaaacaaggt aaggctcttg aatgaaatga catctagtga
      721 aaaattcaaa agcggtacta agaaagatgt ggtcaagttt attgaagact actctagagt
      781 gaatccaaat aagtctgtgt actatttcag tttgaaccac gataaccctg gttggtttta
      841 cttgatgttc aagattaacg caaatagcaa attatacaca tggaatgtga aattaacgaa
      901 cactggttat ttcctggtaa actacaatta tccaagtgtt atccagcttt gtaatggttt
      961 taagacgctt ctaaaatcta acagtagtaa gaatagaatg aacaactacc gttagtagaa
     1021 gggcgagctc agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     1081 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1141 tgctgaaagg aggaactata tccggatatc ccgcaagagg cccggcagta ccggcataac
     1201 caagcctatg cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt
     1261 agatttcata cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta
     1321 aagctagctt atcgatgata agctgtcaaa catgagaatt aattcttgaa gacgaaaggg
     1381 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     1441 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1501 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1561 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1621 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1681 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1741 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1801 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     1861 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     1921 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1981 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2041 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2101 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2161 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2221 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2281 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2341 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2401 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2461 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2521 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2581 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2641 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2701 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     2761 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2821 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2881 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2941 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3001 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3061 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3121 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3181 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3241 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3301 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3361 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3421 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     3481 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     3541 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3601 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     3661 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     3721 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     3781 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     3841 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
     3901 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
     3961 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
     4021 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
     4081 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
     4141 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
     4201 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga
     4261 tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg
     4321 agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc
     4381 gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc
     4441 ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac
     4501 aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc
     4561 ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa gagccgcgag
     4621 cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa
     4681 cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc
     4741 tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat
     4801 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     4861 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     4921 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     4981 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     5041 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     5101 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5161 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     5221 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     5281 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     5341 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     5401 tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     5461 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     5521 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     5581 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     5641 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     5701 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     5761 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     5821 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     5881 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     5941 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     6001 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     6061 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     6121 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     6181 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     6241 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     6301 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
     6361 gccgcaagga atggtgca
//
