LOCUS       40924_25361        4563 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector for high-level expression in insect cells of proteins with a 
            cleavable N-terminal GST-6xHis-S-Tag cassette plus a C-terminal HSV 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4563)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4563)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4563
                     /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"
     CDS             1090..1740
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
                     ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
                     VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
                     MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             1768..1785
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             1795..1839
                     /codon_start=1
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /translation="KETAAAKFERQHMDS"
     CDS             1855..1872
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     CDS             1891..1905
                     /codon_start=1
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
                     /translation="DDDDK"
     CDS             2020..2052
                     /codon_start=1
                     /label=HSV tag
                     /note="HSV (herpes simplex virus) epitope tag"
                     /translation="QPELAPEDPED"
     terminator      2080..2387
                     /label=IE1 terminator
                     /note="terminator of the ie1 gene from the baculovirus 
                     Autographa californica"
     promoter        2417..2521
                     /label=AmpR promoter
     CDS             2522..3379
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3553..4141
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4429..4450
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4465..4495
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4503..4519
                     /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)."
     primer_bind     4527..4543
                     /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 tcacgtacgt ccctcgtgtt 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 accatggcat cccctatact aggttattgg aaaattaagg gccttgtgca acccactcga
     1141 cttcttttgg aatatcttga agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt
     1201 gataaatggc gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct tccttattat
     1261 attgatggtg atgttaaatt aacacagtct atggccatca tacgttatat agctgacaag
     1321 cacaacatgt tgggtggttg tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg
     1381 gttttggata ttagatacgg tgtttcgaga attgcatata gtaaagactt tgaaactctc
     1441 aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt
     1501 cataaaacat atttaaatgg tgatcatgta acccatcctg acttcatgtt gtatgacgct
     1561 cttgatgttg ttttatacat ggacccaatg tgcctggatg cgttcccaaa attagtttgt
     1621 tttaaaaaac gtattgaagc tatcccacaa attgataagt acttgaaatc cagcaagtat
     1681 atagcatggc ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa
     1741 tcggatggtt caactagtgg ttctggtcat caccatcacc atcactcttc tggtaaagaa
     1801 accgctgctg cgaaatttga acgccagcac atggactcgc caccgccttc tggtctggtc
     1861 ccccggggca gcgcaggttc tggtacgatt gatgacgacg acaagagtcc gggcttctcc
     1921 tcaacggatc ccaattggga gctcgtgtac acggcgcgcc tgcaggtcga cggtaccggt
     1981 tcgaagcttg cggccgcaca gctgtataca cgtgcaagcc agccagaact cgccccggaa
     2041 gaccccgagg atctcgagca ctaagtgatt aacctcaggt tatacatata ttttgaattt
     2101 aattaattat acatatattt tatattattt ttgtctttta ttatcgaggg gccgttgttg
     2161 gtgtggggtt ttgcatagaa ataacaatgg gagttggcga cgttgctgcg ccaacaccac
     2221 ctcccttccc tcctttcatc atgtatctgt agataaaata aaatattaaa cctaaaaaca
     2281 agaccgcgcc tatcaacaaa atgataggca ttaacttgcc gctgacgctg tcactaacgt
     2341 tggacgattt gccgactaaa ccttcatcgc ccagtaacca atctagacgt caggtggcac
     2401 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2461 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2521 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2581 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2641 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     2701 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2761 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2821 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2881 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2941 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3001 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3061 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3121 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3181 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3241 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3301 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3361 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3421 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3481 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3541 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3601 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3661 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3721 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3781 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3841 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3901 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3961 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4021 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4081 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4141 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4201 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     4261 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     4321 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     4381 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     4441 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     4501 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgaattccc
     4561 ggg
//
