LOCUS       pET-19b.        5717 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for inducible expression of N-terminally 
            10xHis-tagged proteins with an enterokinase site.
ACCESSION   .
VERSION     .
KEYWORDS    pET-19b
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5717)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5717)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..38
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     terminator      complement(213..260)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    319..335
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(336..350)
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             complement(369..395)
                     /label=9xHis
                     /note="9xHis affinity tag"
     CDS             complement(399..401)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(408..430)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(445..469)
                     /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(470..488)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        797..874
                     /label=lacI promoter
     CDS             875..1954
                     /label=lacI
                     /note="lac repressor"
     protein_bind    1970..1991
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3269..3457
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    3562..3704
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3890..4478)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4652..5509)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5510..5614)
                     /label=AmpR promoter
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
      241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
      301 cgggctttgt tagcagccgg atcctcgagc atatgcttgt cgtcgtcgtc gatatggccg
      361 ctgctgtgat gatgatgatg atgatgatga tgatggccca tggtatatct ccttcttaaa
      421 gttaaacaaa attatttcta gaggggaatt gttatccgct cacaattccc ctatagtgag
      481 tcgtattaat ttcgcgggat cgagatctcg atcctctacg ccggacgcat cgtggccggc
      541 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa
      601 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc
      661 cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg
      721 gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga
      781 gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata
      841 gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt
      901 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
      961 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
     1021 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
     1081 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
     1141 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
     1201 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
     1261 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
     1321 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
     1381 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     1441 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
     1501 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
     1561 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
     1621 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc
     1681 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat
     1741 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
     1801 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
     1861 cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     1921 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt
     1981 agctcactca ttaggcaccg ggatctcgac cgatgccctt gagagccttc aacccagtca
     2041 gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta
     2101 tcatgcaact cgtaggacag gtgccggcag cgctctgggt cattttcggc gaggaccgct
     2161 ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt attcggaatc ttgcacgccc
     2221 tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta
     2281 tcgccggcat ggcggccgac gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct
     2341 ggatggcctt ccccattatg attcttctcg cttccggcgg catcgggatg cccgcgttgc
     2401 aggccatgct gtccaggcag gtagatgacg accatcaggg acagcttcaa ggatcgctcg
     2461 cggctcttac cagcctaact tcgatcactg gaccgctgat cgtcacggcg atttatgccg
     2521 cctcggcgag cacatggaac gggttggcat ggattgtagg cgccgcccta taccttgtct
     2581 gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac ctcgacctga atggaagccg
     2641 gcggcacctc gctaacggat tcaccactcc aagaattgga gccaatcaat tcttgcggag
     2701 aactgtgaat gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag
     2761 cagccgcacg cggcgcatct cgggcagcgt tgggtcctgg ccacgggtgc gcatgatcgt
     2821 gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga
     2881 atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc
     2941 aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc
     3001 gccctgcacc attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac
     3061 acctacatct gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg
     3121 ccgcatccat accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc
     3181 atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa
     3241 cagaaatccc ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat
     3301 ggcccgcttt atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc
     3361 ggatgaacag gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag
     3421 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     3481 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     3541 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     3601 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca tatatgcggt
     3661 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct
     3721 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3781 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3841 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3901 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3961 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4021 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4081 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4141 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4201 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4261 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4321 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4381 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4441 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4501 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4561 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4621 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4681 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4741 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4801 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4861 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4921 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt
     4981 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5041 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5101 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5161 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5221 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg
     5281 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5341 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5401 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5461 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5521 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5581 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     5641 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     5701 cctttcgtct tcaagaa
//
