LOCUS       FLAG.PKCepsilon.        7616 bp DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    FLAG.PKCepsilon.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7616)
  AUTHORS   Cenni V, Doppler H, Sonnenburg ED, Maraldi N, Newton AC, Toker A
  TITLE     Regulation of novel protein kinase C epsilon by phosphorylation.
  JOURNAL   Biochem J. 2002 May 1. 363(Pt 3):537-45.
  PUBMED    11964154
REFERENCE   2  (bases 1 to 7616)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7616)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biochem J. 
            2002 May 1. 363(Pt 3):537-45."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(42..61)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     enhancer        233..612
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        613..816
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        861..879
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             917..940
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             941..3148
                     /gene="PRKCE"
                     /label=PRKCE
                     /note="Protein kinase C epsilon type from Homo sapiens. 
                     Accession#: Q02156"
     promoter        complement(3167..3185)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    3211..3435
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3481..3909
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3923..4252
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4319..5110
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5287..5420
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5457..5473)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5481..5497)
                     /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(5505..5535)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5550..5571)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(5688..5705)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5859..6447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6621..7478)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7479..7583)
                     /label=AmpR promoter
ORIGIN
        1 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
       61 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
      121 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
      181 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
      241 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      301 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      361 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      421 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      481 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      541 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      601 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      661 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      721 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      781 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
      841 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccga
      901 gctcggatcc accatggact ataaggacga tgatgacaaa gtagtgttca atggccttct
      961 taagatcaaa atctgcgagg ccgtgagctt gaagcccaca gcctggtcgc tgcgccatgc
     1021 ggtgggaccc cggccgcaca ctttccttct cgacccctac attgccctca atgtggacga
     1081 ctcgcgcatc ggccaaacgg ccaccaagca gaagaccaac agcccggcct ggcacgacga
     1141 gttcgtcacc gatgtgtgca acggacgcaa gatcgagctg gctgtctttc acgatgcccc
     1201 cataggctac gacgacttcg tggccaactg caccatccag tttgaggagc tgctgcagaa
     1261 cgggagccgc cacttcgagg actggattga tctggagcca gaaggaagag tgtatgtgat
     1321 catcgatctc tcagggtcgt cgggtgaagc ccctaaagac aatgaagagc gtgtgttcag
     1381 ggaacgcatg cggccgagga agcggcaggg ggccgtcagg cgcagggtcc atcaggtcaa
     1441 cggccacaag ttcatggcca cctatcttcg gcagcccacc tactgctccc attgcagaga
     1501 cttcatctgg ggtgtcatag gaaagcaggg ataccagtgt caagtctgca cctgcgtggt
     1561 ccacaagcgg tgccacgagc tcataatcac aaagtgtgct gggttaaaga agcaggagac
     1621 ccccgaccag gtgggctccc agcggttcag cgtcaacatg ccccacaagt tcggtatcca
     1681 caactacaag gtccctacct tctgcgatca ctgtgggtcc ctgctctggg gactcttgcg
     1741 gcagggtttg cagtgtaaag tctgcaaaat gaatgttcac cgtcgatgtg agaccaacgt
     1801 ggctcccaac tgtggagtgg atgccagagg aatcgccaaa gtactggccg acctgggcgt
     1861 taccccagac aaaatcacca acagcggcca gagaaggaaa aagctcattg ctggtgccga
     1921 gtccccgcag cctgcttctg gaagctcacc atctgaggaa gatcgatcca agtcagcacc
     1981 cacctcccct tgtgaccagg aaataaaaga acttgagaac aacattcgga aagccttgtc
     2041 atttgacaac cgaggagagg agcaccgggc agcatcgtct cctgatggcc agctgatgag
     2101 ccccggtgag aatggcgaag tccggcaagg ccaggccaag cgcctgggcc tggatgagtt
     2161 caacttcatc aaggtgttgg gcaaaggcag ctttggcaag gtcatgttgg cagaactcaa
     2221 gggcaaagat gaagtatatg ctgtgaaggt cttaaagaag gacgtcatcc ttcaggatga
     2281 tgacgtggac tgcacaatga cagagaagag gattttggct ctggcacgga aacacccgta
     2341 ccttacccaa ctctactgct gcttccagac caaggaccgc ctctttttcg tcatggaata
     2401 tgtaaatggt ggagacctca tgtttcagat tcagcgctcc cgaaaattcg acgagcctcg
     2461 ttcacggttc tatgctgcag aggtcacatc ggccctcatg ttcctccacc agcatggagt
     2521 catctacagg gatttgaaac tggacaacat ccttctggat gcagaaggtc actgcaagct
     2581 ggctgacttc gggatgtgca aggaagggat tctgaatggt gtgacgacca ccacgttctg
     2641 tgggactcct gactacatag ctcctgagat cctgcaggag ttggagtatg gcccctccgt
     2701 ggactggtgg gccctggggg tgctgatgta cgagatgatg gctggacagc ctccctttga
     2761 ggccgacaat gaggacgacc tatttgagtc catcctccat gacgacgtgc tgtacccagt
     2821 ctggctcagc aaggaggctg tcagcatctt gaaagctttc atgacgaaga atccccacaa
     2881 gcgcctgggc tgtgtggcat cgcagaatgg cgaggacgcc atcaagcagc acccattctt
     2941 caaagagatt gactgggtgc tcctggagca gaagaagatc aagccaccct tcaaaccacg
     3001 cattaaaacc aaaagagacg tcaataattt tgaccaagac tttacccggg aagagccggt
     3061 actcaccctt gtggacgaag caattgtaaa gcagatcaac caggaggaat tcaaaggttt
     3121 ctcctacttt ggtgaagacc tgatgccctg atctagaggg ccctattcta tagtgtcacc
     3181 taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     3241 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     3301 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     3361 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     3421 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     3481 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3541 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3601 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3661 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3721 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3781 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3841 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3901 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3961 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4021 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4081 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4141 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4201 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     4261 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     4321 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4381 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4441 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4501 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4561 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4621 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4681 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4741 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4801 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4861 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4921 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4981 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5041 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5101 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5161 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5221 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5281 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5341 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5401 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5461 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5521 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5581 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5641 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5701 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5761 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5821 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5881 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5941 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6001 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6061 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6121 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6181 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6241 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6301 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6361 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6421 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6481 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6541 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6601 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6661 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6721 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6781 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6841 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6901 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6961 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7021 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7081 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7141 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7201 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7261 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7321 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7381 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7441 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7501 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7561 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcga
//
