LOCUS       sequence-189300-        4365 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-tagged C-terminal domain of RNA polymerase II.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4365)
  AUTHORS   Janke AM, Seo DH, Rahmanian V, Conicella AE, Mathews KL, Burke KA, 
            Mittal J, Fawzi NL
  TITLE     Lysines in the RNA Polymerase II C-Terminal Domain Contribute to 
            TAF15 Fibril Recruitment.
  JOURNAL   Biochemistry. 2017 Oct 11. doi: 10.1021/acs.biochem.7b00310.
  PUBMED    28945358
REFERENCE   2  (bases 1 to 4365)
  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: 
            "Biochemistry. 2017 Oct 11. doi: 10.1021/acs.biochem.7b00310."
COMMENT     Sequence Label: RNAPII_CTD38-52
FEATURES             Location/Qualifiers
     source          1..4365
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     CDS             105..122
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             123..143
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      520..567
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(790..809)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        876..967
                     /gene="bla"
                     /label=AmpR promoter
     CDS             968..1777
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             1860..2942
                     /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     complement(1879..1898)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      3028..3616
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3517..3536
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3770..3787
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3885..3902
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3886..3902
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt ttgaaggaga tataccatgg gcagcgataa gatccaccac caccatcatc
      121 atgagaatct gtattttcaa ggtcatatgt acactccgac gtccccgaaa tacagcccga
      181 cttcgccgaa atattccccg acctccccga agtatagccc aaccagcccg acgtacagcc
      241 cgaccacccc gaagtacagc ccaacgtcgc cgacgtattc gcctaccagc ccggtttaca
      301 ccccgacctc tccgaaatac tcccctacga gcccgactta ctctccgact agcccgaagt
      361 actctccaac ctccccgacc tacagcccta ccagcccaaa gggcagcacc tattcaccga
      421 cgagcccggg ttactcgccg accagcccta cctacagcct gaccagccca gcgattagcc
      481 cggacgatag cgacgaagag aactaataac tcgagccccc tagcataacc ccttggggcc
      541 tctaaacggg tcttgagggg ttttttgccc ctgagacgcg tcaatcgagt tcgtacctaa
      601 gggcgacacc ccctaattag cccgggcgaa aggcccagtc tttcgactga gcctttcgtt
      661 ttatttgatg cctggcagtt ccctactctc gcatggggag tccccacact accatcggcg
      721 ctacggcgtt tcacttctga gttcggcatg gggtcaggtg ggaccaccgc gctactgccg
      781 ccaggcaaac aaggggtgtt atgagccata ttcaggtata aatgggctcg cgataatgtt
      841 cagaattggt taattggttg taacactgac ccctatttgt ttatttttct aaatacattc
      901 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
      961 gaagaatatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     1021 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     1081 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     1141 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     1201 acttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     1261 gatccccgga aaaacagcgt tccaggtatt agaagaatat cctgattcag gtgaaaatat
     1321 tgttgatgcg ctggcagtgt tcctgcgccg gttgcactcg attcctgttt gtaattgtcc
     1381 ttttaacagc gatcgcgtat ttcgcctcgc tcaggcgcaa tcacgaatga ataacggttt
     1441 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     1501 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     1561 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     1621 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     1681 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     1741 attgcaattt catttgatgc tcgatgagtt tttctaagcg gcgcgccatc gaatggcgca
     1801 aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaata
     1861 tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt
     1921 cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg
     1981 cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt
     2041 cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg
     2101 cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac
     2161 gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg
     2221 ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca
     2281 ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt
     2341 tctcccatga ggacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc
     2401 aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct
     2461 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
     2521 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
     2581 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
     2641 ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa gatagctcat
     2701 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
     2761 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgccag
     2821 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
     2881 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     2941 gactcatgac caaaatccct taacgtgagt tacgcgcgcg tcgttccact gagcgtcaga
     3001 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3061 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3121 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3181 agtgtagccg tagttagccc accacttcaa gaactctgta gcaccgccta catacctcgc
     3241 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3301 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3361 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3421 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3481 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3541 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3601 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3661 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3721 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3781 gagcgaggaa gcggaaggcg agagtaggga actgccaggc atcaaactaa gcagaaggcc
     3841 cctgacggat ggcctttttg cgtttctaca aactctttct gtgttgtaaa acgacggcca
     3901 gtcttaagct cgggccccct gggcggttct gataacgagt aatcgttaat ccgcaaataa
     3961 cgtaaaaacc cgcttcggcg ggttttttta tggggggagt ttagggaaag agcatttgtc
     4021 agaatattta agggcgcctg tcactttgct tgatatatga gaattattta accttataaa
     4081 tgagaaaaaa gcaacgcact ttaaataaga tacgttgctt tttcgattga tgaacaccta
     4141 taattaaact attcatctat tatttatgat tttttgtata tacaatattt ctagtttgtt
     4201 aaagagaatt aagaaaataa atctcgaaaa taataaaggg aaaatcagtt tttgatatca
     4261 aaattataca tgtcaacgat aatacaaaat ataatacaaa ctataagatg ttatcagtat
     4321 ttattatgca tttagaataa attttgtgtc gcccttccgc gaaat
//
