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 //