LOCUS       sequence-267365-        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(36..50,51..56,57..73,74..80)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    56..72
                     /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    88..104
                     /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             123..134
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             141..794
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     752..774
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     protein_bind    816..840
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             840..860
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(1090..1114)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      1136..1230
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1324..1983
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1390..1409)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2048..2134
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2227..3309)
                     /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     3271..3290
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3310..3387)
                     /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(3471..3493)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3579..3719
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3634..3653
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3734..3751)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3905..4493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3985..4004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4664..5455,5456..5524))
                     /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     5287..5306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5525..5629)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5697..5715
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagctta attaacaaca ccatttgtcg agaaatcata aaaaatttat ttgctttgtg
       61 agcggataac aattataata gattcaattg tgagcggata acaatttcac acagaattca
      121 ttaaagagga gaaattacat atgtccccta tactaggtta ttggaaaatt aagggccttg
      181 tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg
      241 agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca
      301 atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt
      361 atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa
      421 tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag
      481 actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg
      541 aagatcgttt atgtcataaa acatatttga atggtgatca tgtaacccat cctgacttca
      601 tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc
      661 caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga
      721 aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg
      781 accatcctcc aaaatcggat ctgggtgctg caggtacaag tttgtacaaa aaagcaggcg
      841 agaatcttta ttttcaggga tcctatagcc cgacgtcgcc gagttattca cctacgtccc
      901 catcatactc ccccacgagc cctagttatt cgccaacttc cccgagctat tccccaacat
      961 cacccagcta tagtcccact tcaccctcct attcacctac gagtccatct tattctccaa
     1021 ccagtccttc gtactcaccc acgtccccat cgtattctcc tacttcccct agctaggcgg
     1081 ccgcctagga cccagctttc ttgtacaaag tggtaagctt aattagctga gcttggactc
     1141 ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag aacgctcggt
     1201 tgccgccggg cgttttttat tggtgagaat ccaagtaaca acaccattta aatggagtgg
     1261 ttacaaatgg agtggttaat taaggctagc ttggcgagat tttcaggagc taaggaagct
     1321 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa
     1381 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg
     1441 gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt
     1501 attcacattc ttgcccgcct gatgaatgct catccggaat ttcgtatggc aatgaaagac
     1561 ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact
     1621 gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata
     1681 tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt
     1741 gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac
     1801 gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa
     1861 ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc
     1921 catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg
     1981 taattttttt aaggcagtta ttggtgccct taaacgcctg gggtaatgac tctctagctt
     2041 gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     2101 tttgtcggtg aacgctctcc tgagtaggac aaatccgccc tctagattac gtgcagtcga
     2161 tgataagctg tcaaacatga gaattgtgcc taatgagtga gctaacttac attaattgcg
     2221 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2281 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
     2341 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
     2401 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
     2461 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac
     2521 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
     2581 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
     2641 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
     2701 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
     2761 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
     2821 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
     2881 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
     2941 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
     3001 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
     3061 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
     3121 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
     3181 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
     3241 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
     3301 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
     3361 aaaggttttg caccattcga tggtgtcgga atttcgggca gcgttgggtc ctggccacgg
     3421 gtgcgcatga tctagagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     3481 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     3541 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     3601 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
     3661 tgcaccacat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     3721 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3781 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3841 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3901 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3961 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4021 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4081 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4141 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4201 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4261 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4321 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     4381 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4441 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4501 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4561 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4621 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4681 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4741 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4801 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4861 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4921 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4981 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5041 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5101 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5161 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5221 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5281 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5341 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5401 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5461 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5521 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5581 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5641 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     5701 ggcgtatcac gaggcccttt cgtcttcac
//
