LOCUS       40924_25366        4629 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector for high-level expression in insect cells of secreted 
            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 4629)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4629)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4629
                     /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"
     sig_peptide     1084..1143
                     /label=AKH signal sequence
                     /note="signal sequence from tobacco hornworm adipokinetic 
                     hormone"
     CDS             1156..1806
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
                     ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
                     VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
                     MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             1834..1851
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             1861..1905
                     /codon_start=1
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /translation="KETAAAKFERQHMDS"
     CDS             1921..1938
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     CDS             1957..1971
                     /codon_start=1
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
                     /translation="DDDDK"
     CDS             2086..2118
                     /codon_start=1
                     /label=HSV tag
                     /note="HSV (herpes simplex virus) epitope tag"
                     /translation="QPELAPEDPED"
     terminator      2146..2453
                     /label=IE1 terminator
                     /note="terminator of the ie1 gene from the baculovirus 
                     Autographa californica"
     promoter        2483..2587
                     /label=AmpR promoter
     CDS             2588..3445
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3619..4207
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4495..4516
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4531..4561
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4569..4585
                     /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     4593..4609
                     /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 actatgtaca agctcacagt cttcctgatg ttcatcgctt tcgtcatcat cgctgaggcc
     1141 cagctgacca tggcatcccc tatactaggt tattggaaaa ttaagggcct tgtgcaaccc
     1201 actcgacttc ttttggaata tcttgaagaa aaatatgaag agcatttgta tgagcgcgat
     1261 gaaggtgata aatggcgaaa caaaaagttt gaattgggtt tggagtttcc caatcttcct
     1321 tattatattg atggtgatgt taaattaaca cagtctatgg ccatcatacg ttatatagct
     1381 gacaagcaca acatgttggg tggttgtcca aaagagcgtg cagagatttc aatgcttgaa
     1441 ggagcggttt tggatattag atacggtgtt tcgagaattg catatagtaa agactttgaa
     1501 actctcaaag ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt
     1561 ttatgtcata aaacatattt aaatggtgat catgtaaccc atcctgactt catgttgtat
     1621 gacgctcttg atgttgtttt atacatggac ccaatgtgcc tggatgcgtt cccaaaatta
     1681 gtttgtttta aaaaacgtat tgaagctatc ccacaaattg ataagtactt gaaatccagc
     1741 aagtatatag catggccttt gcagggctgg caagccacgt ttggtggtgg cgaccatcct
     1801 ccaaaatcgg atggttcaac tagtggttct ggtcatcacc atcaccatca ctcttctggt
     1861 aaagaaaccg ctgctgcgaa atttgaacgc cagcacatgg actcgccacc gccttctggt
     1921 ctggtccccc ggggcagcgc aggttctggt acgattgatg acgacgacaa gagtccgggc
     1981 ttctcctcaa cggatcccaa ttgggagctc gtgtacacgg cgcgcctgca ggtcgacggt
     2041 accggttcga agcttgcggc cgcacagctg tatacacgtg caagccagcc agaactcgcc
     2101 ccggaagacc ccgaggatct cgagcactaa gtgattaacc tcaggttata catatatttt
     2161 gaatttaatt aattatacat atattttata ttatttttgt cttttattat cgaggggccg
     2221 ttgttggtgt ggggttttgc atagaaataa caatgggagt tggcgacgtt gctgcgccaa
     2281 caccacctcc cttccctcct ttcatcatgt atctgtagat aaaataaaat attaaaccta
     2341 aaaacaagac cgcgcctatc aacaaaatga taggcattaa cttgccgctg acgctgtcac
     2401 taacgttgga cgatttgccg actaaacctt catcgcccag taaccaatct agacgtcagg
     2461 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2521 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2581 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     2641 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2701 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     2761 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     2821 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2881 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2941 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3001 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3061 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3121 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     3181 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     3241 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     3301 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     3361 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     3421 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     3481 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3541 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3601 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3661 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3721 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     3781 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3841 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3901 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3961 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4021 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4081 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4141 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4201 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4261 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     4321 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     4381 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     4441 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     4501 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     4561 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacga
     4621 attcccggg
//
