LOCUS       sequence-272883-        5729 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses GST-tagged 10xCTD heptapeptide repeat in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5729)
  AUTHORS   Castaneda AF, Didychuk AL, Louder RK, McCollum CO, Davis ZH, Nogales
            E, Glaunsinger BA
  TITLE     The gammaherpesviral TATA-box-binding protein directly interacts 
            with the CTD of host RNA Pol II to direct late gene transcription.
  JOURNAL   PLoS Pathog. 2020 Sep 4;16(9):e1008843. doi: 
            10.1371/journal.ppat.1008843. eCollection 2020 Sep.
  PUBMED    32886723
REFERENCE   2  (bases 1 to 5729)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Pathog."; date: "2020-09-4"; volume: "16(9):e1008843. doi"; pages: "
            10.1371/journal.ppat.1008843. eCollection 2020 Sep"
COMMENT     Sequence Label: pQLink-GST-10xCTD
FEATURES             Location/Qualifiers
     source          1..5729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..15,16..21,22..38,39..45)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    21..37
                     /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)."
     protein_bind    53..69
                     /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             88..99
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             106..759
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     717..739
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     protein_bind    781..805
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             805..825
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(1055..1079)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      1101..1195
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1289..1948
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1355..1374)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2013..2099
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2192..3274)
                     /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     3236..3255
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3275..3352)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(3436..3458)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3544..3684
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3599..3618
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3699..3716)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3870..4458)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3950..3969)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4629..5420,5421..5489))
                     /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     5252..5271
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5490..5594)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5662..5680
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc aattgtgagc
       61 ggataacaat ttcacacaga attcattaaa gaggagaaat tacatatgtc ccctatacta
      121 ggttattgga aaattaaggg ccttgtgcaa cccactcgac ttcttttgga atatcttgaa
      181 gaaaaatatg aagagcattt gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag
      241 tttgaattgg gtttggagtt tcccaatctt ccttattata ttgatggtga tgttaaatta
      301 acacagtcta tggccatcat acgttatata gctgacaagc acaacatgtt gggtggttgt
      361 ccaaaagagc gtgcagagat ttcaatgctt gaaggagcgg ttttggatat tagatacggt
      421 gtttcgagaa ttgcatatag taaagacttt gaaactctca aagttgattt tcttagcaag
      481 ctacctgaaa tgctgaaaat gttcgaagat cgtttatgtc ataaaacata tttgaatggt
      541 gatcatgtaa cccatcctga cttcatgttg tatgacgctc ttgatgttgt tttatacatg
      601 gacccaatgt gcctggatgc gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct
      661 atcccacaaa ttgataagta cttgaaatcc agcaagtata tagcatggcc tttgcagggc
      721 tggcaagcca cgtttggtgg tggcgaccat cctccaaaat cggatctggg tgctgcaggt
      781 acaagtttgt acaaaaaagc aggcgagaat ctttattttc agggatccta tagcccgacg
      841 tcgccgagtt attcacctac gtccccatca tactccccca cgagccctag ttattcgcca
      901 acttccccga gctattcccc aacatcaccc agctatagtc ccacttcacc ctcctattca
      961 cctacgagtc catcttattc tccaaccagt ccttcgtact cacccacgtc cccatcgtat
     1021 tctcctactt cccctagcta ggcggccgcc taggacccag ctttcttgta caaagtggta
     1081 agcttaatta gctgagcttg gactcctgtt gatagatcca gtaatgacct cagaactcca
     1141 tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag
     1201 taacaacacc atttaaatgg agtggttaca aatggagtgg ttaattaagg ctagcttggc
     1261 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
     1321 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
     1381 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
     1441 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
     1501 ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
     1561 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
     1621 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
     1681 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
     1741 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
     1801 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
     1861 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
     1921 ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac
     1981 gcctggggta atgactctct agcttgaggc atcaaataaa acgaaaggct cagtcgaaag
     2041 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     2101 cgccctctag attacgtgca gtcgatgata agctgtcaaa catgagaatt gtgcctaatg
     2161 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2221 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2281 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
     2341 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
     2401 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
     2461 ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
     2521 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
     2581 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
     2641 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
     2701 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
     2761 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     2821 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     2881 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     2941 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     3001 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     3061 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     3121 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
     3181 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
     3241 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
     3301 ccgggcgcta tcatgccata ccgcgaaagg ttttgcacca ttcgatggtg tcggaatttc
     3361 gggcagcgtt gggtcctggc cacgggtgcg catgatctag agctgcctcg cgcgtttcgg
     3421 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     3481 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     3541 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg
     3601 gcatcagagc agattgtact gagagtgcac cacatgcggt gtgaaatacc gcacagatgc
     3661 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3721 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3781 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3841 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3901 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3961 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4021 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4081 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4141 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4201 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4261 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     4321 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4381 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4441 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4501 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4561 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4621 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4681 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4741 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4801 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4861 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4921 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4981 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5041 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5101 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5161 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5221 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5281 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5341 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5401 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5461 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5521 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5581 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     5641 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct tcacctcgag
     5701 cttaattaac aacaccattt gtcgagaaa
//
