LOCUS       pBiEx-2.        6162 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector for high-level expression in both bacterial and insect cells 
            of proteins with a cleavable N-terminal GST-6xHis-S-Tag cassette 
            plus a C-terminal HSV tag.
ACCESSION   .
VERSION     .
KEYWORDS    pBiEx-2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6162)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6162)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6162
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..483
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     promoter        487..1078
                     /label=IE1 promoter
                     /note="promoter of the ie1 gene from the baculovirus
                     Autographa californica"
     promoter        1092..1110
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1111..1135
                     /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)."
     RBS             1150..1172
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1185..1835
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
     CDS             1872..1889
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             1899..1943
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             1959..1976
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             1995..2009
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     misc_feature    2008..2115
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             2121..2153
                     /label=HSV tag
                     /note="HSV (herpes simplex virus) epitope tag"
     terminator      2223..2270
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2295..2602
                     /label=IE1 terminator
                     /note="terminator of the ie1 gene from the baculovirus 
                     Autographa californica"
     protein_bind    complement(2690..2711)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2727..3806)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(3807..3884)
                     /label=lacI promoter
     promoter        4016..4120
                     /label=AmpR promoter
     CDS             4121..4978
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5152..5740
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6028..6049
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6064..6094
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     6126..6142
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgcgtaaaac acaatcaagt atgagtcata agctgatgtc atgttttgca cacggctcat
       61 aaccgaactg gctttacgag tagaattcta cttgtaacgc acgatcagtg gatgatgtca
      121 tttgtttttc aaatcgagat gatgtcatgt tttgcacacg gctcataaac tcgctttacg
      181 agtagaattc tacgtgtaac gcacgatcga ttgatgagtc atttgttttg caatatgata
      241 tcatacaata tgactcattt gtttttcaaa accgaacttg atttacgggt agaattctac
      301 ttgtaaagca caatcaaaaa gatgatgtca tttgtttttc aaaactgaac tcgctttacg
      361 agtagaattc tacgtgtaaa acacaatcaa gaaatgatgt catttgttat aaaaataaaa
      421 gctgatgtca tgttttgcac atggctcata actaaactcg ctttacgggt agaattctac
      481 gcgcgtcgat gtctttgtga tgcgcgcgac atttttgtag gttattgata aaatgaacgg
      541 atacgttgcc cgacattatc attaaatcct tggcgtagaa tttgtcgggt ccattgtccg
      601 tgtgcgctag catgcccgta acggacctcg tacttttggc ttcaaaggtt ttgcgcacag
      661 acaaaatgtg ccacacttgc agctctgcat gtgtgcgcgt taccacaaat cccaacggcg
      721 cagtgtactt gttgtatgca aataaatctc gataaaggcg cggcgcgcga atgcagctga
      781 tcacgtacgc tcctcgtgtt ccgttcaagg acggtgttat cgacctcaga ttaatgttta
      841 tcggccgact gttttcgtat ccgctcacca aacgcgtttt tgcattaaca ttgtatgtcg
      901 gcggatgttc tatatctaat ttgaataaat aaacgataac cgcgttggtt ttagagggca
      961 taataaaaga aatattgtta tcgtgttcgc cattagggca gtataaattg acgttcatgt
     1021 tggatattgt ttcagttgca agttgacact ggcggcgaca agatcgtgaa caaccaagtg
     1081 accgcggaaa ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct
     1141 agaaataatt ttgtttaact ttaagaagga gatataccat ggcatcccct atactaggtt
     1201 attggaaaat taagggcctt gtgcaaccca ctcgacttct tttggaatat cttgaagaaa
     1261 aatatgaaga gcatttgtat gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg
     1321 aattgggttt ggagtttccc aatcttcctt attatattga tggtgatgtt aaattaacac
     1381 agtctatggc catcatacgt tatatagctg acaagcacaa catgttgggt ggttgtccaa
     1441 aagagcgtgc agagatttca atgcttgaag gagcggtttt ggatattaga tacggtgttt
     1501 cgagaattgc atatagtaaa gactttgaaa ctctcaaagt tgattttctt agcaagctac
     1561 ctgaaatgct gaaaatgttc gaagatcgtt tatgtcataa aacatattta aatggtgatc
     1621 atgtaaccca tcctgacttc atgttgtatg acgctcttga tgttgtttta tacatggacc
     1681 caatgtgcct ggatgcgttc ccaaaattag tttgttttaa aaaacgtatt gaagctatcc
     1741 cacaaattga taagtacttg aaatccagca agtatatagc atggcctttg cagggctggc
     1801 aagccacgtt tggtggtggc gaccatcctc caaaatcgga tggttcaact agtggttctg
     1861 gttccatggc acaccatcac caccatcact cttctggtaa agaaaccgct gctgcgaaat
     1921 ttgaacgcca gcacatggac tcgccaccgc cttctggtct ggtcccccgg ggcagcgcag
     1981 gttctggtac gattgatgac gacgacaaga tctctggccc gggctcggat cccaattggg
     2041 agctcgtgta cacggcgcgc ctgcaggtcg acggtaccgg ttcgaagctt gcggccgcac
     2101 agctgtatac acgtgcaagc cagccagaac tcgccccgga agaccccgag gatctcgagc
     2161 actaagtgat taacctcagc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     2221 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     2281 gaactatcct caggttatac atatattttg aatttaatta attatacata tattttatat
     2341 tatttttgtc ttttattatc gaggggccgt tgttggtgtg gggttttgca tagaaataac
     2401 aatgggagtt ggcgacgttg ctgcgccaac accacctccc ttccctcctt tcatcatgta
     2461 tctgtagata aaataaaata ttaaacctaa aaacaagacc gcgcctatca acaaaatgat
     2521 aggcattaac ttgccgctga cgctgtcact aacgttggac gatttgccga ctaaaccttc
     2581 atcgcccagt aaccaatcta gacgtcacga tagtcatgcc ccgcgcccac cggaaggagc
     2641 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     2701 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2761 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     2821 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     2881 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     2941 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     3001 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     3061 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     3121 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     3181 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     3241 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     3301 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     3361 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     3421 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     3481 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     3541 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     3601 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     3661 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     3721 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     3781 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     3841 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     3901 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     3961 cgccgccgca aggaatggtg catggacgtc aggtggcact tttcggggaa atgtgcgcgg
     4021 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4081 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4141 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4201 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4261 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4321 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4381 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4441 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4501 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4561 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4621 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4681 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4741 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4801 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4861 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4921 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4981 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5041 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5101 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5161 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5221 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5281 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5341 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5401 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5461 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5521 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5581 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5641 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5701 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5761 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5821 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     5881 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     5941 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     6001 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     6061 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     6121 tcacacagga aacagctatg accatgatta cgaattcccg gg
//
