LOCUS       pIVEX1.3WG                3236 bp ds-DNA     circular SYN
DEFINITION  Cloning vector pIVEX1.3WG (C-terminal His6-tag)
ACCESSION   .
VERSION     .
KEYWORDS    pIVEX1.3WG, RTS, cell-free protein synthesis
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3236)
  AUTHORS   biotechrabbit GmbH
            Volmerstr. 9, 12489 Berlin, Germany
COMMENT     pIVEX1.3WG vector for high-level expression of His6-tagged proteins
            in the cell-free RTS Wheat Germ CECF system. Cloning vector with
            C-terminal His6-tag.
            Ordering information: 
            BR1401301  RTS pIVEX Wheat Germ His6-tag Vector Set
FEATURES             Location/Qualifiers
     source          1..3236
                     /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=IRES
                     /note="IRES (internal ribosome entry site) element 
                     optimized for the RTS Wheat Germ system"
     misc_feature    249..294
                     /label=MCS
                     /note="multiple cloning site for the insertion of the 
                     target gene"
     CDS             251..253
                     /codon_start=1
                     /label=Start
                     /note="start of translation"
                     /translation="M"
     CDS             304..321
                     /codon_start=1
                     /product="6×His affinity tag"
                     /label=6×His
                     /translation="HHHHHH"
     misc_feature    322..324
                     /label=Stop
                     /note="stop codon"
     3'UTR           335..580
                     /label=3'-Enhancer
                     /note="expression enhancing element derived from TMV 
                     (Tobacco mosaic virus)"
     primer_bind     complement(594..611)
                     /label=M13-fwd
                     /note="3' sequencing primer"
     rep_origin      complement(752..1207)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1234..1338
                     /gene="bla"
                     /label=ampR promoter
     CDS             1339..2199
                     /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      2370..2958
                     /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 atggcacata tgagcggccg cgtcgactcg agcgagctcc cggggggggt
      301 tctcatcatc atcatcatca ttaataaggt acccagctct tctggtttgg tttggacctc
      361 tggtcctgca acttgaggta gtcaagatgc ataataaata acggattgtg tccgtaatca
      421 cacgtggtgc gtacgataac gcatagtgtt tttccctcca cttaaatcga agggttgtgt
      481 cttggatcgc gcgggtcaaa tgtatatggt tcatatacat ccgcaggcac gtaataaagc
      541 gaggggttcg aatccccccg ttacccccgg taggggccca tatatgcgga attcactggc
      601 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
      661 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
      721 ccaacagttg cgcagcctga atggcgaatg gaaattgtaa gcgttaatat tttgttaaaa
      781 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
      841 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
      901 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
      961 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     1021 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     1081 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     1141 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     1201 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1261 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1321 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1381 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1441 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1501 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1561 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     1621 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1681 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1741 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     1801 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     1861 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     1921 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     1981 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2041 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2101 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2161 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2221 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2281 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2341 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2401 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2461 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     2521 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2581 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2641 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2701 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2761 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     2821 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     2881 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     2941 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3001 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3061 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3121 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     3181 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcg
//
