LOCUS       pIVEX2.3d                3560 bp ds-DNA     circular SYN
DEFINITION  Cloning vector pIVEX2.3d (C-terminal His6-tag)
ACCESSION   .
VERSION     .
KEYWORDS    pIVEX2.3d; RTS; cell-free protein synthesis
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3560)
  AUTHORS   biotechrabbit GmbH
            Volmerstr. 9, 12489 Berlin, Germany
COMMENT     pIVEX2.3d vector for high-level expression of His6-tagged proteins
            in the cell-free RTS E. coli system. Cloning vector with
            C-terminal His6-tag.
            Ordering information: 
            BR1400701  RTS pIVEX E. coli His-tag 2nd Generation Vector Set
FEATURES             Location/Qualifiers
     source          1..3560
                     /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)"
     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             295..303
                     /codon_start=1
                     /label=Linker
                     /translation="GGS"
     CDS             304..321
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    322..324
                     /label=Stop
     primer_bind     complement(415..433)
                     /label=T7-rev
                     /note="3' sequencing primer"
     terminator      429..476
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(1293..1881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2052..2912)
                     /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(2913..3017)
                     /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 atggcacata tgagcggccg cgtcgactcg agcgagctcc cggggggggt
      301 tctcatcatc atcatcatca ttaataaaag ggcgaattcc agcacactgg cggccgttac
      361 tagtggatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
      421 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
      481 aaggaggaac tatatccgga tatccacagg acgggtgtgg tcgccatgat cgcgtagtcg
      541 atagtggctc caagtagcga agcgagcagg actgggcggc ggccaaagcg gtcggacagt
      601 gctccgagaa cgggtgcgca tagaaattgc atcaacgcat atagcgctag cagcacgcca
      661 tagtgactgg cgatgctgtc ggaatggacg atatcccgca agaggcccgg cagtaccggc
      721 ataaccaagc ctatgcctac agcatccagg gtgacggtgc cgaggatgac gatgagcgca
      781 ttgttagatt tcatacacgg tgcctgactg cgttagcaat ttaactgtga taaactaccg
      841 cattaaagct tatcgatgat aagctgtcaa acatgagaat tcgtaatcat gtcatagctg
      901 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
      961 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     1021 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     1081 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     1141 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1201 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1261 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1321 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1381 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1441 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1501 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1561 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1621 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1681 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     1741 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1801 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1861 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1921 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1981 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2041 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2101 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2161 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2221 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2281 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2341 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2401 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2461 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2521 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2581 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2641 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2701 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2761 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2821 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2881 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2941 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3001 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     3061 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     3121 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     3181 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     3241 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatata
     3301 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     3361 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     3421 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     3481 agtcacgacg ttgtaaaacg acggccagtg ccaagcttgc atgcaaggag atggcgccca
     3541 acagtccccc ggccacgggg
//
