LOCUS       40924_21519        5772 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Bacterial expression vector pGEXGSTp65(285-551), complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5772)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5772)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 5772)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5772)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 
            9052, BELGIUM"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5772
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(274..447)
                     /codon_start=1
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     misc_feature    complement(448..485)
                     /label=5' UTR of lacZ, incl. RBS
                     /note="5' UTR of lacZ, incl. RBS"
     protein_bind    467..483
                     /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)."
     promoter        complement(491..521)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(536..557)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(573..1652)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(1653..1730)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     rep_origin      complement(1974..2562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2736..3593)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3594..3698)
                     /label=AmpR promoter
     CDS             complement(4031..4402)
                     /codon_start=1
                     /label=RelA (p65) AD
                     /note="transcriptional activation domain of human RelA,
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     CDS             complement(4841..4852)
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     CDS             complement(4862..5515)
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     protein_bind    complement(5538..5554)
                     /label=lac operator
                     /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)."
     promoter        complement(5562..5590)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
ORIGIN
        1 aattccaacg ccatcaaaaa taattcgcgt ctggccttcc tgtagccagc tttcatcaac
       61 attaaatgtg agcgagtaac aacccgtcgg attctccgtg ggaacaaacg gcggattgac
      121 cgtaatggga taggttacgt tggtgtagat gggcgcatcg taaccgtgca tctgccagtt
      181 tgaggggacg acgacagtat cggcctcagg aagatcgcac tccagccagc tttccggcac
      241 cgcttctggt gccggaaacc aggcaaagcg ccattcgcca ttcaggctgc gcaactgttg
      301 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
      361 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
      421 ggccagtgaa tccgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
      481 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
      541 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
      601 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
      661 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
      721 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
      781 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
      841 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
      901 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
      961 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     1021 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     1081 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     1141 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     1201 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     1261 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     1321 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     1381 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     1441 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     1501 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     1561 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     1621 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     1681 gggcgctatc atgccatacc gcgaaaggtt ttgcaccatt cgatggtgtc ggaatttatg
     1741 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
     1801 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1861 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1921 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1981 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2041 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2101 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2161 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2221 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2281 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2341 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2401 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2461 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2521 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2581 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2641 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2701 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2761 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2821 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2881 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2941 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3001 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg
     3061 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3121 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3181 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3241 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3301 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     3361 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3421 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3481 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3541 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3601 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3661 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3721 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     3781 aggccctttc gtcttcaaga attatacact ccgctatcgc tacgtgactg ggtcatggct
     3841 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     3901 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3961 tcatcaccga aacgcgcgag gcagatcgtc agtcagtcac gatgcggccg ctcgagtcgt
     4021 caccccctta ggagctgatc tgactcagca gggctgagaa gtccatgtcc gcaatggagg
     4081 agaagtcttc atctcctgaa aggaggccat tggggagccc cggggccccc agtggagcag
     4141 gagctgggtc ggggggcctc tgggcccctg tcactaggcg agttatagcc tcagggtact
     4201 ccatcagcat gggctcagtt gtgtgggggg ccacaggtat gccctggttc agcagctgct
     4261 gaaactcgga gttgtcgacg gatgccaggt ctgtgaacac agctgggtct gtgctgttgc
     4321 caagcaaggc ccccaggtct tcatcatcaa actgcagctg cagcagggcc tctgacagcg
     4381 ttccttcccc agcctgggtg ggcttggggg caggtggggc cacagcctga ggagggcctg
     4441 gggctaggac tgggacaggg gctggggcct gggccagagc tgataccatg gctggagcag
     4501 gggcaggggc tggagcctgg ggcaggactt ggggaggggc cggggccaag gccgaggcct
     4561 ggctgatctg cccagaagga aacaccatgg tgggaaactc atcatagttg atggtgctca
     4621 gggatgacgt aaagggatag ggctggggtg ctggcttggg gacagaagct gagctgcggg
     4681 aaggcacagc aatgcgtcga ggtggaggcc gggggtcggt gggtccgctg aaaggactct
     4741 tcttcatgat gctcttgaag gtctcatatg tccttttacg tttctcctca atccggtgac
     4801 gatcgtctgt atctggcagg tactggaatt cggggatccc acgaccttcg atcagatccg
     4861 attttggagg atggtcgcca ccaccaaacg tggcttgcca gccctgcaaa ggccatgcta
     4921 tatacttgct ggatttcaag tacttatcaa tttgtgggat agcttcaata cgttttttaa
     4981 aacaaactaa ttttgggaac gcatccaggc acattgggtc catgtataaa acaacatcaa
     5041 gagcgtcata caacatgaag tcaggatggg ttacatgatc accatttaaa tatgttttat
     5101 gacataaacg atcttcgaac attttcagca tttcaggtag cttgctaaga aaatcaactt
     5161 tgagagtttc aaagtcttta ctatatgcaa ttctcgaaac accgtatcta atatccaaaa
     5221 ccgctccttc aagcattgaa atctctgcac gctcttttgg acaaccaccc aacatgttgt
     5281 gcttgtcagc tatataacgt atgatggcca tagactgtgt taatttaaca tcaccatcaa
     5341 tataataagg aagattggga aactccaaac ccaattcaaa ctttttgttt cgccatttat
     5401 caccttcatc gcgctcatac aaatgctctt catatttttc ttcaagatat tccaaaagaa
     5461 gtcgagtggg ttgcacaagg cccttaattt tccaataacc tagtataggg gacatgaata
     5521 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acattatacg agccgatgat
     5581 taattgtcaa cagctcattt cagaatattt gccagaaccg ttatgatgtc ggcgcaaaaa
     5641 acattatcca gaacgggagt gcgccttgag cgacacgaat tatgcagtga tttacgacct
     5701 gcacagccat accacagctt ccgatggctg cctgacgcca gaagcattgg tgcaccgtgc
     5761 agtcgataag ct
//
