LOCUS       40924_24613        7106 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pHis-p65f285-551, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7106)
  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  JOURNAL   Unpublished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 7106)
  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 7106)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7106)
  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            "Unpublished"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..7106
                     /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             1463..1834
                     /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    1838..2627
                     /label=3UTR
                     /note="3UTR"
     polyA_signal    2703..2927
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2973..3401
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3415..3744
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3811..4602
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4779..4912
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4949..4965)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4973..4989)
                     /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(4997..5027)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5042..5063)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5351..5939)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6113..6970)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6971..7075)
                     /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 aggtaccagg
     1021 atccagtgtg gtggaattcc agtacctgcc agatacagac gatcgtcacc ggattgagga
     1081 gaaacgtaaa aggacatatg agaccttcaa gagcatcatg aagaagagtc ctttcagcgg
     1141 acccaccgac ccccggcctc cacctcgacg cattgctgtg ccttcccgca gctcagcttc
     1201 tgtccccaag ccagcacccc agccctatcc ctttacgtca tccctgagca ccatcaacta
     1261 tgatgagttt cccaccatgg tgtttccttc tgggcagatc agccaggcct cggccttggc
     1321 cccggcccct ccccaagtcc tgccccaggc tccagcccct gcccctgctc cagccatggt
     1381 atcagctctg gcccaggccc cagcccctgt cccagtccta gccccaggcc ctcctcaggc
     1441 tgtggcccca cctgccccca agcccaccca ggctggggaa ggaacgctgt cagaggccct
     1501 gctgcagctg cagtttgatg atgaagacct gggggccttg cttggcaaca gcacagaccc
     1561 agctgtgttc acagacctgg catccgtcga caactccgag tttcagcagc tgctgaacca
     1621 gggcatacct gtggcccccc acacaactga gcccatgctg atggagtacc ctgaggctat
     1681 aactcgccta gtgacagggg cccagaggcc ccccgaccca gctcctgctc cactgggggc
     1741 cccggggctc cccaatggcc tcctttcagg agatgaagac ttctcctcca ttgcggacat
     1801 ggacttctca gccctgctga gtcagatcag ctcctaaggg ggtgacgcct gccctcccca
     1861 gagcactggt tgcaggggat tgaagccctc caaaagcact tacggattct ggtggggtgt
     1921 gttccaactg cccccaactt tgtggatgtc ttccttggag gggggagcca tattttattc
     1981 ttttattgtc agtatctgta tctctctctc tttttggagg tgcttaagca gaagcattaa
     2041 cttctctgga aaggggggag ctggggaaac tcaaactttt cccctgtcct gatggtcagc
     2101 tcccttctct gtagggaact gtggggtccc ccatccccat cctccagctt ctggtactct
     2161 cctagagaca gaagcaggct ggaggtaagg cctttgagcc cacaaagcct tatcaagtgt
     2221 cttccatcat ggattcatta cagcttaatc aaaataacgc cccagatacc agcccctgta
     2281 tggcactggc attgtccctg tgcctaacac cagcgtttga ggggctgcct tcctgcccta
     2341 cagaggtctc tgccggctct ttccttgctc aaccatggct gaaggaaaca gtgcaacagc
     2401 actggctctc tccaggatcc agaaggggtt tggtctggac ttccttgctc tcccctcttc
     2461 tcaagtgcct taatagtagg gtaagttgtt aagagtgggg gagagcaggc tggcagctct
     2521 ccagtcagga ggcatagttt ttagtgaaca atcaaagcac ttggactctt gctctttcta
     2581 ctctgaacta ataaagctgt tgccaagctg aaaaaaaaaa aaaaaggaat tctgcagata
     2641 tccagcacag tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc
     2701 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2761 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2821 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2881 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2941 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3001 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3061 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3121 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3181 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3241 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3301 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3361 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3421 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3481 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3541 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3601 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3661 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3721 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3781 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3841 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3901 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3961 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4021 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4081 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4141 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4201 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4261 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4321 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4381 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4441 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4501 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4561 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4621 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4681 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4741 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4801 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4861 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4921 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4981 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5041 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5101 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5161 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5221 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5281 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5341 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5401 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5461 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5521 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5581 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5641 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5701 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5761 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5821 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5881 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5941 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6001 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6061 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6121 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6181 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6241 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6301 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6361 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6421 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6481 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6541 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6601 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6661 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6721 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6781 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6841 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6901 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6961 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7021 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7081 ttccccgaaa agtgccacct gacgtc
//
