LOCUS pIVEX1.4WG 3287 bp ds-DNA circular SYN DEFINITION Cloning vector pIVEX1.4WG (N-terminal His6-tag) ACCESSION . VERSION . KEYWORDS pIVEX1.4WG; RTS; cell-free protein synthesis SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3287) AUTHORS biotechrabbit GmbH Volmerstr. 9, 12489 Berlin, Germany COMMENT pIVEX1.4WG vector for high-level expression of His6-tagged proteins in the cell-free RTS Wheat Germ CECF system. Cloning vector with cleavable N-terminal His6-tag. Ordering information: BR1401301 RTS pIVEX Wheat Germ His6-tag Vector Set FEATURES Location/Qualifiers source 1..3287 /organism="synthetic DNA construct" /mol_type="circular plasmid" primer_bind 108..125 /label=M13-rev /note="5' sequencing primer" promoter 145..163 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" 5'UTR 177..232 /label=5'-Enhancer /note="IRES (internal ribosome entry site) element optimized for the RTS Wheat Germ system" CDS 251..253 /codon_start=1 /label=Start /note="start of translation" /translation="M" CDS 263..280 /codon_start=1 /product="6×His affinity tag" /label=6×His /translation="HHHHHH" CDS 290..301 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" misc_feature 302..365 /label=MCS /note="multiple cloning site for the insertion of the target gene" misc_feature 368..370 /label=Stop /note="stop codon" 3'UTR 386..628 /label=3'-Enhancer /note="expression enhancing element derived from TMV (Tobacco mosaic virus)" primer_bind complement(645..662) /label=M13-fwd /note="3' sequencing primer" rep_origin complement(803..1258) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 1285..1389 /gene="bla" /label=ampR promoter CDS 1390..2250 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=ampR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 2421..3009 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct 61 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta 121 tgaccatgat tacgccaagc tcattaatac gactcactat aggcctaagc ttacaaatac 181 tcccccacaa cagcttacaa tactccccca cacagcttac aaatactccc ccacaacagc 241 ttgtcgaacc atgtctggtt ctcatcatca tcatcatcat agcagcggca tcgaaggccg 301 cggccgctta attaaacata tgaccatggc aagtcgactc gagcgagctc tgcagcccgg 361 gatccggtaa ctaactaagg tacccagctc ttctggtttg gtttggacct ctggtcctgc 421 aacttgaggt agtcaagatg cataataaat aacggattgt gtccgtaatc acacgtggtg 481 cgtacgataa cgcatagtgt ttttccctcc acttaaatcg aagggttgtg tcttggatcg 541 cgcgggtcaa atgtatatgg ttcatataca tccgcaggca cgtaataaag cgaggggttc 601 gaatcccccc gttacccccg gtaggggccc atatatgcgg aattcactgg ccgtcgtttt 661 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc 721 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt 781 gcgcagcctg aatggcgaat ggaaattgta agcgttaata ttttgttaaa attcgcgtta 841 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat 901 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca 961 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc 1021 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta 1081 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg 1141 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg 1201 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca 1261 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat 1321 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa 1381 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt 1441 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag 1501 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt 1561 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg 1621 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag 1681 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta 1741 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg 1801 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta 1861 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac 1921 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt 1981 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca 2041 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag 2101 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta 2161 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag 2221 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt 2281 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat 2341 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta 2401 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa 2461 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt 2521 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag 2581 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta 2641 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca 2701 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag 2761 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa 2821 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga 2881 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc 2941 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc 3001 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt 3061 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt 3121 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag 3181 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa 3241 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcg //