LOCUS       pIVEX2.4d                3583 bp ds-DNA     circular SYN
DEFINITION  Cloning vector pIVEX2.4d (N-terminal His6-tag)
ACCESSION   .
VERSION     .
KEYWORDS    pIVEX2.4d; RTS; cell-free protein synthesis
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3583)
  AUTHORS   biotechrabbit GmbH
            Volmerstr. 9, 12489 Berlin, Germany
COMMENT     pIVEX2.4d vector for high-level expression of His6-tagged proteins
            in the cell-free RTS E. coli system. Cloning vector with
            cleavable N-terminal His6-tag.
            Ordering information: 
            BR1400701  RTS pIVEX E. coli His-tag 2nd Generation Vector Set
FEATURES             Location/Qualifiers
     source          1..3583
                     /organism="synthetic DNA construct"
                     /mol_type="circular plasmid"
     primer_bind     172..191
                     /label=T7-fwd
                     /note="5' sequencing primer"
     promoter        172..190
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             222..244
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             251..253
                     /codon_start=1
                     /label=Start
                     /note="start of translation"
                     /translation="M"
     CDS             263..280
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /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
     primer_bind     complement(438..456)
                     /label=T7-rev
                     /note="3' sequencing primer"
     terminator      452..499
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(1316..1904)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2075..2935)
                     /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"
     promoter        complement(2936..3040)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
       61 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
      121 ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac
      181 tcactatagg gagaccacaa cggtttccct ctagaaataa ttttgtttaa ctttaagaag
      241 gagatatacc atgtctggtt ctcatcatca tcatcatcat agcagcggca tcgaaggccg
      301 cggccgctta attaaacata tgaccatggc aagtcgactc gagcgagctc tgcagcccgg
      361 gatccggtaa ctaactaaga tccggtaaga tccggctgct aacaaagccc gaaaggaagc
      421 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
      481 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatatccac aggacgggtg
      541 tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc aggactgggc
      601 ggcggccaaa gcggtcggac agtgctccga gaacgggtgc gcatagaaat tgcatcaacg
      661 catatagcgc tagcagcacg ccatagtgac tggcgatgct gtcggaatgg acgatatccc
      721 gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg
      781 tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc
      841 aatttaactg tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag
      901 aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
      961 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     1021 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     1081 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     1141 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     1201 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     1261 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1321 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1381 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1441 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1501 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1561 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1621 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     1681 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     1741 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     1801 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1861 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1921 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1981 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2041 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2101 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2161 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     2221 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     2281 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     2341 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     2401 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     2461 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     2521 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     2581 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     2641 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     2701 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     2761 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     2821 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     2881 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     2941 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3001 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3061 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     3121 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     3181 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     3241 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     3301 ttgtactgag agtgcaccat atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     3361 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     3421 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     3481 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
     3541 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg ggg
//
