LOCUS       pBiEx-3.        5373 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector for high-level expression in both bacterial and insect cells 
            of proteins with a C-terminal S-Tag-8xHis cassette.
ACCESSION   .
VERSION     .
KEYWORDS    pBiEx-3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5373)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5373)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5373
                     /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)"
     misc_feature    1177..1273
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             1179..1181
                     /codon_start=1
                     /label=ATG
                     /note="ATG"
                     /translation="M"
     CDS             1284..1328
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             1350..1373
                     /label=8xHis
                     /note="8xHis affinity tag"
     terminator      1434..1481
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1506..1813
                     /label=IE1 terminator
                     /note="terminator of the ie1 gene from the baculovirus 
                     Autographa californica"
     protein_bind    complement(1901..1922)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(1938..3017)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(3018..3095)
                     /label=lacI promoter
     promoter        3227..3331
                     /label=AmpR promoter
     CDS             3332..4189
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4363..4951
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5239..5260
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5275..5305
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     5337..5353
                     /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 ggcatcgtta acacgtcaag
     1201 agctcgcgga tcccaattgg cagatctcgg cgcgcctgca ggtcgacggt accggttcga
     1261 agcttgcggc cgcatcttct ggtaaagaaa ccgctgctgc gaaatttgaa cgccagcaca
     1321 tggactcgcc accgccttct ggcctcgagc accaccatca ccatcaccat cactaagtga
     1381 ttaacctcag ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     1441 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatcc
     1501 tcaggttata catatatttt gaatttaatt aattatacat atattttata ttatttttgt
     1561 cttttattat cgaggggccg ttgttggtgt ggggttttgc atagaaataa caatgggagt
     1621 tggcgacgtt gctgcgccaa caccacctcc cttccctcct ttcatcatgt atctgtagat
     1681 aaaataaaat attaaaccta aaaacaagac cgcgcctatc aacaaaatga taggcattaa
     1741 cttgccgctg acgctgtcac taacgttgga cgatttgccg actaaacctt catcgcccag
     1801 taaccaatct agacgtcacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt
     1861 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     1921 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1981 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
     2041 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
     2101 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
     2161 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
     2221 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
     2281 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
     2341 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
     2401 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
     2461 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
     2521 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
     2581 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
     2641 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
     2701 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
     2761 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
     2821 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
     2881 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
     2941 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
     3001 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     3061 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     3121 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     3181 aaggaatggt gcatggacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3241 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3301 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     3361 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     3421 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     3481 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     3541 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     3601 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     3661 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     3721 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     3781 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     3841 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     3901 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     3961 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4021 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     4081 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4141 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     4201 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4261 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4321 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4381 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4441 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4501 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4561 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4621 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4681 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4741 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4801 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4861 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4921 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4981 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5041 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     5101 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     5161 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     5221 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     5281 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     5341 aaacagctat gaccatgatt acgaattccc ggg
//
