LOCUS       40924_36423        7087 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Mammalian expression vector pRcRSVp65S311A, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7087)
  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  JOURNAL   Unpublished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 7087)
  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 7087)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7087)
  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            "Unpublished"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..7087
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        340..602
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    1630..1632
                     /label=S311A
                     /note="S311A"
     CDS             1981..2352
                     /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    2356..2541
                     /label=3' UTR of hNF-kappaB p65 subunit
                     /note="3' UTR of hNF-kappaB p65 subunit"
     polyA_signal    2588..2812
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2858..3371
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3399..3729
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3796..4587
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4764..4898
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4930..4946)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4954..4970)
                     /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(4978..5008)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5023..5044)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5332..5920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6094..6951)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6952..7056)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtatctgag
      241 gggactaggg tgtgtttagg cgaaaagcgg ggcttcggtt gtacgcggtt aggagtcccc
      301 tcaggatata gtagtttcgc ttttgcatag ggagggggaa atgtagtctt atgcaataca
      361 cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag gagagaaaaa
      421 gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat taggaaggca
      481 acagacaggt ctgacatgga ttggacgaac cactgaattc cgcattgcag agataattgt
      541 atttaagtgc ctagctcgat acaataaacg ccatttgacc attcaccaca ttggtgtgca
      601 cctccaagct tgatatcgaa ttccggagtt ccatggctcg tctgtagtgc acgccgcggg
      661 cccagctgcg accccggccc cgcccccggg accccggcca tggacgaact gttccccctc
      721 atcttcccgg cagagccagc ccaggcctct ggcccctatg tggagatcat tgagcagccc
      781 aagcagcggg gcatgcgctt ccgctacaag tgcgaggggc gctccgcggg cagcatccca
      841 ggcgagagga gcacagatac caccaagacc caccccacca tcaagatcaa tggctacaca
      901 ggaccaggga cagtgcgcat ctccctggtc accaaggacc ctcctcaccg gcctcacccc
      961 cacgagcttg taggaaagga ctgccgggat ggcttctatg aggctgagct ctgcccggac
     1021 cgctgcatcc acagtttcca gaacctggga atccagtgtg tgaagaagcg ggacctggag
     1081 caggctatca gtcagcgcat ccagaccaac aacaacccct tccaagttcc tatagaagag
     1141 cagcgtgggg actacgacct gaatgctgtg cggctctgct tccaggtgac agtgcgggac
     1201 ccatcaggca ggcccctccg cctgccgcct gtccttcctc atcccatctt tgacaatcgt
     1261 gcccccaaca ctgccgagct caagatctgc cgagtgaacc gaaactctgg cagctgcctc
     1321 ggtggggatg agatcttcct actgtgtgac aaggtgcaga aagaggacat tgaggtgtat
     1381 ttcacgggac caggctggga ggcccgaggc tccttttcgc aagctgatgt gcaccgacaa
     1441 gtggccattg tgttccggac ccctccctac gcagacccca gcctgcaggc tcctgtgcgt
     1501 gtctccatgc agctgcggcg gccttccgac cgggagctca gtgagcccat ggaattccag
     1561 tacctgccag atacagacga tcgtcaccgg attgaggaga aacgtaaaag gacatatgag
     1621 accttcaagg ccatcatgaa gaagagtcct ttcagcggac ccaccgaccc ccggcctcca
     1681 cctcgacgca ttgctgtgcc ttcccgcagc tcagcttctg tccccaagcc agcaccccag
     1741 ccctatccct ttacgtcatc cctgagcacc atcaactatg atgagtttcc caccatggtg
     1801 tttccttctg ggcagatcag ccaggcctcg gccttggccc cggcccctcc ccaagtcctg
     1861 ccccaggctc cagcccctgc ccctgctcca gccatggtat cagctctggc ccaggcccca
     1921 gcccctgtcc cagtcctagc cccaggccct cctcaggctg tggccccacc tgcccccaag
     1981 cccacccagg ctggggaagg aacgctgtca gaggccctgc tgcagctgca gtttgatgat
     2041 gaagacctgg gggccttgct tggcaacagc acagacccag ctgtgttcac agacctggca
     2101 tccgtcgaca actccgagtt tcagcagctg ctgaaccagg gcatacctgt ggccccccac
     2161 acaactgagc ccatgctgat ggagtaccct gaggctataa ctcgcctagt gacaggggcc
     2221 cagaggcccc ccgacccagc tcctgctcca ctgggggccc cggggctccc caatggcctc
     2281 ctttcaggag atgaagactt ctcctccatt gcggacatgg acttctcagc cctgctgagt
     2341 cagatcagct cctaaggggg tgacgcctgc cctccccaga gcactggttg caggggattg
     2401 aagccctcca aaagcactta cggattctgg tggggtgtgt tccaactgcc cccaactttg
     2461 tggatgtctt ccttggaggg gggagccata ttttattctt ttattgtcag tatctgtatc
     2521 tctctctctt tttggaggtg cttaaggccg ctcgagcatg catctagagc tcgctgatca
     2581 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2641 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2701 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2761 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     2821 gcggaaagaa ccagctgggg ctcgaggggg gatccccacg cgccctgtag cggcgcatta
     2881 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2941 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3001 gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3061 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3121 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3181 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc
     3241 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3301 acgtttacaa tttaaatatt tgcttataca atcttcctgt ttttggggct tttctgatta
     3361 tcaaccgggg tgggtaccga gctcgaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3421 agtccccagg ctccccaggc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3481 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3541 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3601 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3661 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3721 ttttgcaaaa agctcccggg agcttggata tccattttcg gatctgatca agagacagga
     3781 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3841 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3901 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3961 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4021 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4081 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4141 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4201 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4261 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4321 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4381 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4441 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4501 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4561 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4621 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4681 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4741 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4801 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4861 tgtccaaact catcaatgta tcttatcatg tctggatccc gtcgacctcg agagcttggc
     4921 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4981 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5041 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5101 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5161 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5221 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5281 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5341 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5401 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5461 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5521 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5581 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5641 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5701 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5761 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5821 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5881 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5941 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6001 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6061 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6121 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6181 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6241 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6301 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6361 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6421 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6481 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6541 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6601 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6661 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6721 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6781 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6841 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6901 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6961 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7021 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7081 tgacgtc
//
