LOCUS       40924_24598        7940 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pHis-p65, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7940)
  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  JOURNAL   Unpublished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 7940)
  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 7940)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7940)
  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            "Unpublished"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..7940
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             932..949
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             953..985
                     /codon_start=1
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
                     /translation="MASMTGGQQMG"
     CDS             989..1012
                     /codon_start=1
                     /label=Xpress(TM) tag
                     /note="Xpress(TM) epitope tag, including an enterokinase 
                     recognition and cleavage site"
                     /translation="DLYDDDDK"
     CDS             2297..2668
                     /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"
     misc_feature    2672..3461
                     /label=3' UTR of hNF-kappaB p65 subunit
                     /note="3' UTR of hNF-kappaB p65 subunit"
     polyA_signal    3537..3761
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3807..4235
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4249..4578
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4645..5436
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5613..5746
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5783..5799)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5807..5823)
                     /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(5831..5861)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5876..5897)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6185..6773)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6947..7804)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7805..7909)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttacca tggggggttc tcatcatcat catcatcatg gtatggctag
      961 catgactggt ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggtaccgga
     1021 cgaactgttc cccctcatct tcccggcaga gccagcccag gcctctggcc cctatgtgga
     1081 gatcattgag cagcccaagc agcggggcat gcgcttccgc tacaagtgcg aggggcgctc
     1141 cgcgggcagc atcccaggcg agaggagcac agataccacc aagacccacc ccaccatcaa
     1201 gatcaatggc tacacaggac cagggacagt gcgcatctcc ctggtcacca aggaccctcc
     1261 tcaccggcct cacccccacg agcttgtagg aaaggactgc cgggatggct tctatgaggc
     1321 tgagctctgc ccggaccgct gcatccacag tttccagaac ctgggaatcc agtgtgtgaa
     1381 gaagcgggac ctggagcagg ctatcagtca gcgcatccag accaacaaca accccttcca
     1441 agttcctata gaagagcagc gtggggacta cgacctgaat gctgtgcggc tctgcttcca
     1501 ggtgacagtg cgggacccat caggcaggcc cctccgcctg ccgcctgtcc ttcctcatcc
     1561 catctttgac aatcgtgccc ccaacactgc cgagctcaag atctgccgag tgaaccgaaa
     1621 ctctggcagc tgcctcggtg gggatgagat cttcctactg tgtgacaagg tgcagaaaga
     1681 ggacattgag gtgtatttca cgggaccagg ctgggaggcc cgaggctcct tttcgcaagc
     1741 tgatgtgcac cgacaagtgg ccattgtgtt ccggacccct ccctacgcag accccagcct
     1801 gcaggctcct gtgcgtgtct ccatgcagct gcggcggcct tccgaccggg agctcagtga
     1861 gcccatggaa ttccagtacc tgccagatac agacgatcgt caccggattg aggagaaacg
     1921 taaaaggaca tatgagacct tcaagagcat catgaagaag agtcctttca gcggacccac
     1981 cgacccccgg cctccacctc gacgcattgc tgtgccttcc cgcagctcag cttctgtccc
     2041 caagccagca ccccagccct atccctttac gtcatccctg agcaccatca actatgatga
     2101 gtttcccacc atggtgtttc cttctgggca gatcagccag gcctcggcct tggccccggc
     2161 ccctccccaa gtcctgcccc aggctccagc ccctgcccct gctccagcca tggtatcagc
     2221 tctggcccag gccccagccc ctgtcccagt cctagcccca ggccctcctc aggctgtggc
     2281 cccacctgcc cccaagccca cccaggctgg ggaaggaacg ctgtcagagg ccctgctgca
     2341 gctgcagttt gatgatgaag acctgggggc cttgcttggc aacagcacag acccagctgt
     2401 gttcacagac ctggcatccg tcgacaactc cgagtttcag cagctgctga accagggcat
     2461 acctgtggcc ccccacacaa ctgagcccat gctgatggag taccctgagg ctataactcg
     2521 cctagtgaca ggggcccaga ggccccccga cccagctcct gctccactgg gggccccggg
     2581 gctccccaat ggcctccttt caggagatga agacttctcc tccattgcgg acatggactt
     2641 ctcagccctg ctgagtcaga tcagctccta agggggtgac gcctgccctc cccagagcac
     2701 tggttgcagg ggattgaagc cctccaaaag cacttacgga ttctggtggg gtgtgttcca
     2761 actgccccca actttgtgga tgtcttcctt ggagggggga gccatatttt attcttttat
     2821 tgtcagtatc tgtatctctc tctctttttg gaggtgctta agcagaagca ttaacttctc
     2881 tggaaagggg ggagctgggg aaactcaaac ttttcccctg tcctgatggt cagctccctt
     2941 ctctgtaggg aactgtgggg tcccccatcc ccatcctcca gcttctggta ctctcctaga
     3001 gacagaagca ggctggaggt aaggcctttg agcccacaaa gccttatcaa gtgtcttcca
     3061 tcatggattc attacagctt aatcaaaata acgccccaga taccagcccc tgtatggcac
     3121 tggcattgtc cctgtgccta acaccagcgt ttgaggggct gccttcctgc cctacagagg
     3181 tctctgccgg ctctttcctt gctcaaccat ggctgaagga aacagtgcaa cagcactggc
     3241 tctctccagg atccagaagg ggtttggtct ggacttcctt gctctcccct cttctcaagt
     3301 gccttaatag tagggtaagt tgttaagagt gggggagagc aggctggcag ctctccagtc
     3361 aggaggcata gtttttagtg aacaatcaaa gcacttggac tcttgctctt tctactctga
     3421 actaataaag ctgttgccaa gctgaaaaaa aaaaaaaaag gaattctgca gatatccagc
     3481 acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     3541 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3601 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3661 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3721 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3781 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3841 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3901 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3961 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4021 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4081 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4141 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4201 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4261 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4321 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4381 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4441 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4501 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4561 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4621 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4681 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4741 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4801 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4861 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4921 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4981 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5041 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5101 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5161 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5221 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5281 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5341 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5401 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5461 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5521 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5581 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5641 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5701 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5761 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5821 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5881 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5941 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6001 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6061 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6121 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6181 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6241 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6301 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6361 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6421 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6481 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6541 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6601 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6661 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6721 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6781 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6841 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6901 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6961 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7021 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7081 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7141 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7201 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7261 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7321 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7381 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7441 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7501 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7561 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7621 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7681 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7741 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7801 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7861 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7921 gaaaagtgcc acctgacgtc
//
