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
//
