LOCUS       40924_16555        7046 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Express RelA in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7046)
  AUTHORS   Burstein E, Hoberg JE, Wilkinson AS, Rumble JM, Csomos RA, Komarck 
            CM, Maine GN, Wilkinson JC, Mayo MW, Duckett CS
  TITLE     COMMD proteins, a novel family of structural and functional homologs
            of MURR1.
  JOURNAL   J Biol Chem. 2005 Jun 10;280(23):22222-32. Epub 2005 Mar 30.
  PUBMED    15799966
REFERENCE   2  (bases 1 to 7046)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7046)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol
            Chem."; date: "2005-06-10"; volume: "280(23)"; pages: "22222-32.
            Epub 2005 Mar 30"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7046
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        252..462
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        558..1739
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             1804..1830
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             3124..3495
                     /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"
     primer_bind     complement(4180..4196)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4409..4864
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4881..4900)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     5000..5022
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5060..5078)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        5146..5250
                     /label=AmpR promoter
     CDS             5251..6108
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6282..6870
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7024..7041
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttc
      241 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
      301 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
      361 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
      421 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctttgca
      481 aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct tctaggtctt
      541 gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
      601 ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg
      661 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
      721 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
      781 aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc
      841 ttgcgtgcct tgaattactt ccacgcccct ggctgcagta cgtgattctt gatcccgagc
      901 ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc
      961 gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc
     1021 ttcgcgcctg tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg
     1081 ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg
     1141 gtatttcggt ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt
     1201 cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg
     1261 gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa
     1321 ggctggcccg gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg
     1381 cagggagctc aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac
     1441 aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg
     1501 cgccgtccag gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg
     1561 gggaggggtt ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc
     1621 cagcttggca cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt
     1681 tcattctcaa gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgag
     1741 gaattctcta gagatccctc gacctcgaga tccattgtgc tggatctcac catggcttct
     1801 agctatcctt atgacgtgcc tgactatgcc agcctgggat ccatggacga actgttcccc
     1861 ctcatcttcc cggcagagcc agcccaggcc tctggcccct atgtggagat cattgagcag
     1921 cccaagcagc ggggcatgcg cttccgctac aagtgcgagg ggcgctccgc gggcagcatc
     1981 ccaggcgaga ggagcacaga taccaccaag acccacccca ccatcaagat caatggctac
     2041 acaggaccag ggacagtgcg catctccctg gtcaccaagg accctcctca ccggcctcac
     2101 ccccacgagc ttgtaggaaa ggactgccgg gatggcttct atgaggctga gctctgcccg
     2161 gaccgctgca tccacagttt ccagaacctg ggaatccagt gtgtgaagaa gcgggacctg
     2221 gagcaggcta tcagtcagcg catccagacc aacaacaacc ccttccaagt tcctatagaa
     2281 gagcagcgtg gggactacga cctgaatgct gtgcggctct gcttccaggt gacagtgcgg
     2341 gacccatcag gcaggcccct ccgcctgccg cctgtccttt ctcatcccat ctttgacaat
     2401 cgtgccccca acactgccga gctcaagatc tgccgagtga accgaaactc tggcagctgc
     2461 ctcggtgggg atgagatctt cctactgtgt gacaaggtgc agaaagagga cattgaggtg
     2521 tatttcacgg gaccaggctg ggaggcccga ggctcctttt cgcaagctga tgtgcaccga
     2581 caagtggcca ttgtgttccg gacccctccc tacgcagacc ccagcctgca ggctcctgtg
     2641 cgtgtctcca tgcagctgcg gcggccttcc gaccgggagc tcagtgagcc catggaattc
     2701 cagtacctgc cagatacaga cgatcgtcac cggattgagg agaaacgtaa aaggacatat
     2761 gagaccttca agagcatcat gaagaagagt cctttcagcg gacccaccga cccccggcct
     2821 ccacctcgac gcattgctgt gccttcccgc agctcagctt ctgtccccaa gccagcaccc
     2881 cagccctatc cctttacgtc atccctgagc accatcaact atgatgagtt tcccaccatg
     2941 gtgtttcctt ctgggcagat cagccaggcc tcggccttgg ccccggcccc tccccaagtc
     3001 ctgccccagg ctccagcccc tgcccctgct ccagccatgg tatcagctct ggcccaggcc
     3061 ccagcccctg tcccagtcct agccccaggc cctcctcagg ctgtggcccc acctgccccc
     3121 aagcccaccc aggctgggga aggaacgctg tcagaggccc tgctgcagct gcagtttgat
     3181 gatgaagacc tgggggcctt gcttggcaac agcacagacc cagctgtgtt cacagacctg
     3241 gcatccgtcg acaactccga gtttcagcag ctgctgaacc agggcatacc tgtggccccc
     3301 cacacaactg agcccatgct gatggagtac cctgaggcta taactcgcct agtgacaggg
     3361 gcccagaggc cccccgaccc agctcctgct ccactggggg ccccggggct ccccaatggc
     3421 ctcctttcag gagatgaaga cttctcctcc attgcggaca tggacttctc agccctgctg
     3481 agtcagatca gctcctaagc ggccgcgact ctagagtgag ggtccccacc tgggaccctt
     3541 gagagtatca ggtctcccac gtgggagaca agaaatccct gtttaatatt taaacagcag
     3601 tgttccccat ctgggtcctt gcacccctca ctctggcctc agccgactgc acagcggccc
     3661 ctgcatcccc ttggctgtga ggcccctgga caagcagagg tggccagagc tgggaggcat
     3721 ggccctgggg tcccacgaat ttgctgggga atctcgtttt tcttcttaag acttttggga
     3781 catggtttga ctcccgaaca tcaccgacgc gtctcctgtt tttctgggtg gcctcgggac
     3841 acctgccctg cccccacgag ggtcaggact gtgactcttt ttagggccag gcaggtgcct
     3901 ggacatttgc cttgctggac ggggactggg gatgtgggag ggagcagaca ggaggaatca
     3961 tgtcaggcct gtgtgtgaaa ggaagctcca ctgtcaccct ccacctcttc accccccact
     4021 caccagtgtc ccctccactg tcacattgta actgaacttc aggataataa agtgtttgcc
     4081 tccaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     4141 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaagaattca ctggccgtcg ttttacaacg
     4201 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     4261 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     4321 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4381 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     4441 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4501 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4561 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4621 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4681 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4741 cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4801 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     4861 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4921 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4981 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     5041 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5101 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5161 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5221 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     5281 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5341 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5401 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5461 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5521 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5581 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5641 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5701 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5761 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5821 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5881 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5941 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6001 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6061 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6121 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6181 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     6241 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     6301 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6361 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6421 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6481 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6541 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6601 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6661 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6721 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6781 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6841 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6901 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6961 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     7021 cgcagcgagt cagtgagcga ggaagc
//
