LOCUS       pET-27b(+).        5414 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector with a signal sequence and a kanamycin resistance 
            marker for expression of C-terminally HSV-tagged proteins in the 
            periplasm.
ACCESSION   .
VERSION     .
KEYWORDS    pET-27b(+).
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5414)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5414)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(140..157)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(164..196)
                     /label=HSV tag
                     /note="HSV (herpes simplex virus) epitope tag"
     misc_feature    212..279
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     sig_peptide     complement(278..343)
                     /label=pelB signal sequence
                     /note="leader peptide for secretion"
     RBS             complement(351..373)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(388..412)
                     /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(413..431)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        740..817
                     /label=lacI promoter
     CDS             818..1897
                     /label=lacI
                     /note="lac repressor"
     protein_bind    1913..1934
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2709..2897
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    3002..3144
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3330..3918)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             4040..4852
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(4948..5403)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gacatcctcg gggtcttccg
      181 gggcgagttc tggctggcta gcccgtttga tctcgagtgc ggccgcaagc ttgtcgacgg
      241 agctcgaatt cggatccgaa ttaattccga tatccatggc catcgccggc tgggcagcga
      301 ggagcagcag accagcagca gcggtcggca gcaggtattt catatgtata tctccttctt
      361 aaagttaaac aaaattattt ctagagggga attgttatcc gctcacaatt cccctatagt
      421 gagtcgtatt aatttcgcgg gatcgagatc tcgatcctct acgccggacg catcgtggcc
      481 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg
      541 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca
      601 ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg
      661 gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag
      721 ggagagcgtc gagatcccgg acaccatcga atggcgcaaa acctttcgcg gtatggcatg
      781 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
      841 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
      901 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
      961 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     1021 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     1081 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     1141 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     1201 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     1261 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     1321 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     1381 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     1441 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     1501 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     1561 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
     1621 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac
     1681 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     1741 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
     1801 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1861 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa
     1921 gttagctcac tcattaggca ccgggatctc gaccgatgcc cttgagagcc ttcaacccag
     1981 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     2041 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
     2101 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg
     2161 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
     2221 ttatcgccgg catggcggcc ccacgggtgc gcatgatcgt gctcctgtcg ttgaggaccc
     2281 ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata cgcgagcgaa
     2341 cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga atggtcttcg
     2401 gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc attatgttcc
     2461 ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct gtattaacga
     2521 agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat accgccagtt
     2581 gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc cgtatcgtga
     2641 gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc ccttacacgg
     2701 aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc
     2761 agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct
     2821 gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg
     2881 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2941 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     3001 gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc
     3061 atcagagcag attgtactga gagtgcacca tatatgcggt gtgaaatacc gcacagatgc
     3121 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3181 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3241 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3301 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3361 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3421 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3481 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3541 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3601 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3661 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3721 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     3781 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3841 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3901 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3961 aacgaaaact cacgttaagg gattttggtc atgaacaata aaactgtctg cttacataaa
     4021 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct ctaggccgcg
     4081 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     4141 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
     4201 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     4261 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     4321 atggttactc accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc
     4381 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     4441 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     4501 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     4561 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     4621 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     4681 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     4741 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     4801 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaagaatt
     4861 aattcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4921 gcacatttcc ccgaaaagtg ccacctgaaa ttgtaaacgt taatattttg ttaaaattcg
     4981 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     5041 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     5101 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     5161 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     5221 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     5281 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     5341 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     5401 cgtcccattc gcca
//
