LOCUS       pBiEx-1.        5475 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 6xHis-S-Tag cassette plus a 
            C-terminal HSV tag.
ACCESSION   .
VERSION     .
KEYWORDS    pBiEx-1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5475)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5475)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5475
                     /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             1179..1181
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             1185..1202
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             1212..1256
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             1272..1289
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             1308..1322
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     misc_feature    1321..1428
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             1434..1466
                     /label=HSV tag
                     /note="HSV (herpes simplex virus) epitope tag"
     terminator      1536..1583
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1608..1915
                     /label=IE1 terminator
                     /note="terminator of the ie1 gene from the baculovirus 
                     Autographa californica"
     protein_bind    complement(2003..2024)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2040..3119)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(3120..3197)
                     /label=lacI promoter
     promoter        3329..3433
                     /label=AmpR promoter
     CDS             3434..4291
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4465..5053
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5341..5362
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5377..5407
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     5439..5455
                     /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 ggcacaccat caccaccatc
     1201 actcttctgg taaagaaacc gctgctgcga aatttgaacg ccagcacatg gactcgccac
     1261 cgccttctgg tctggtcccc cggggcagcg caggttctgg tacgattgat gacgacgaca
     1321 agatctctgg cccgggctcg gatcccaatt gggagctcgt gtacacggcg cgcctgcagg
     1381 tcgacggtac cggttcgaag cttgcggccg cacagctgta tacacgtgca agccagccag
     1441 aactcgcccc ggaagacccc gaggatctcg agcactaagt gattaacctc agccgaaagg
     1501 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     1561 aacgggtctt gaggggtttt ttgctgaaag gaggaactat cctcaggtta tacatatatt
     1621 ttgaatttaa ttaattatac atatatttta tattattttt gtcttttatt atcgaggggc
     1681 cgttgttggt gtggggtttt gcatagaaat aacaatggga gttggcgacg ttgctgcgcc
     1741 aacaccacct cccttccctc ctttcatcat gtatctgtag ataaaataaa atattaaacc
     1801 taaaaacaag accgcgccta tcaacaaaat gataggcatt aacttgccgc tgacgctgtc
     1861 actaacgttg gacgatttgc cgactaaacc ttcatcgccc agtaaccaat ctagacgtca
     1921 cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     1981 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac
     2041 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2101 cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg
     2161 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
     2221 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
     2281 gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg
     2341 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
     2401 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
     2461 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
     2521 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
     2581 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
     2641 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
     2701 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
     2761 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
     2821 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
     2881 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
     2941 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
     3001 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
     3061 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
     3121 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
     3181 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
     3241 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatggac
     3301 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3361 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3421 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3481 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3541 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3601 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3661 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3721 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3781 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3841 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3901 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3961 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4021 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4081 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4141 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4201 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4261 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4321 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4381 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4441 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4501 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4561 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     4621 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4681 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4741 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4801 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4861 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4921 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4981 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5041 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5101 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5161 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     5221 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     5281 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     5341 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     5401 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     5461 ttacgaattc ccggg
//
