LOCUS       pIVEX1.4WG-GUS                5066 bp ds-DNA     circular SYN
DEFINITION  Control Vector GUS
ACCESSION   .
VERSION     .
KEYWORDS    pIVEX1.4WG-GUS; RTS; cell-free protein synthesis
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5066)
  AUTHORS   biotechrabbit GmbH
            Volmerstr. 9, 12489 Berlin, Germany
COMMENT     GUS (beta-glucuronidase) expression vector with cleavable 
            N-terminal 6xHis-tag for the control reaction.
            Ordering information: 
            BR1401001  RTS 100 Wheat Germ CECF Kit
            BR1401101  RTS 500 Wheat Germ CECF Kit
            BR1402501  RTS 100 Wheat Germ Kit
FEATURES             Location/Qualifiers
     source          1..5066
                     /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"
     CDS             251..2149
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase with cleavable N-terminal 
                     6xHis-tag"
                     /label=6xHis-GUS
                     /note="263..280  6xHis affinity tag
                     290..301  Factor Xa recognition and cleavage site"
                     /translation="MSGSHHHHHHSSGIEGRGRLIKHMTMVRPVETPTREIKKLDGLWA
                     FSLDRENCGIDQRWWESALQESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGW
                     AGQRIVLRFDAVTHYGKVWVNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNEL
                     NWQTIPPGMVITDENGKKKQSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDC
                     NHASVDWQVVANGDVSVELRDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTA
                     KSQTECDIYPLRVGIRSVAVKGEQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDH
                     ALMDWIGANSYRTSHYPYAEEMLDWADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKEL
                     YSEEAVNGETQQAHLQAIKELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATR
                     KLDPTRPITCVNVMFCDAHTDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAW
                     QEKLHQPIIITEYGVDTLAGLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNF
                     ADFATSQGILRVGGNKKGIFTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQPGIR"
     primer_bind     370..381
                     /label=M13-fwd
                     /note="3' sequencing primer"
     3'UTR           2165..2407
                     /label=3'-Enhancer
                     /note="expression enhancing element derived from TMV 
                     (Tobacco mosaic virus)"
     primer_bind     complement(2424..2441)
                     /label=M13-fwd
                     /note="3' sequencing primer"
     rep_origin      complement(2582..3037)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3064..3168
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3169..4029
                     /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      4200..4788
                     /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 tgaccatggt ccgtcctgta gaaaccccaa cccgtgaaat
      361 caaaaaactc gacggcctgt gggcattcag tctggatcgc gaaaactgtg gaattgatca
      421 gcgttggtgg gaaagcgcgt tacaagaaag ccgggcaatt gctgtgccag gcagttttaa
      481 cgatcagttc gccgatgcag atattcgtaa ttatgcgggc aacgtctggt atcagcgcga
      541 agtctttata ccgaaaggtt gggcaggcca gcgtatcgtg ctgcgtttcg atgcggtcac
      601 tcattacggc aaagtgtggg tcaataatca ggaagtgatg gagcatcagg gcggctatac
      661 gccatttgaa gccgatgtca cgccgtatgt tattgccggg aaaagtgtac gtatcaccgt
      721 ttgtgtgaac aacgaactga actggcagac tatcccgccg ggaatggtga ttaccgacga
      781 aaacggcaag aaaaagcagt cttacttcca tgatttcttt aactatgccg gaatccatcg
      841 cagcgtaatg ctctacacca cgccgaacac ctgggtggac gatatcaccg tggtgacgca
      901 tgtcgcgcaa gactgtaacc acgcgtctgt tgactggcag gtggtggcca atggtgatgt
      961 cagcgttgaa ctgcgtgatg cggatcaaca ggtggttgca actggacaag gcactagcgg
     1021 gactttgcaa gtggtgaatc cgcacctctg gcaaccgggt gaaggttatc tctatgaact
     1081 gtgcgtcaca gccaaaagcc agacagagtg tgatatctac ccgcttcgcg tcggcatccg
     1141 gtcagtggca gtgaagggcg aacagttcct gattaaccac aaaccgttct actttactgg
     1201 ctttggtcgt catgaagatg cggacttgcg tggcaaagga ttcgataacg tgctgatggt
     1261 gcacgaccac gcattaatgg actggattgg ggccaactcc taccgtacct cgcattaccc
     1321 ttacgctgaa gagatgctcg actgggcaga tgaacatggc atcgtggtga ttgatgaaac
     1381 tgctgctgtc ggctttaacc tctctttagg cattggtttc gaagcgggca acaagccgaa
     1441 agaactgtac agcgaagagg cagtcaacgg ggaaactcag caagcgcact tacaggcgat
     1501 taaagagctg atagcgcgtg acaaaaacca cccaagcgtg gtgatgtgga gtattgccaa
     1561 cgaaccggat acccgtccgc aaggtgcacg ggaatatttc gcgccactgg cggaagcaac
     1621 gcgtaaactc gacccgacgc gtccgatcac ctgcgtcaat gtaatgttct gcgacgctca
     1681 caccgatacc atcagcgatc tctttgatgt gctgtgcctg aaccgttatt acggatggta
     1741 tgtccaaagc ggcgatttgg aaacggcaga gaaggtactg gaaaaagaac ttctggcctg
     1801 gcaggagaaa ctgcatcagc cgattatcat caccgaatac ggcgtggata cgttagccgg
     1861 gctgcactca atgtacaccg acatgtggag tgaagagtat cagtgtgcat ggctggatat
     1921 gtatcaccgc gtctttgatc gcgtcagcgc cgtcgtcggt gaacaggtat ggaatttcgc
     1981 cgattttgcg acctcgcaag gcatattgcg cgttggcggt aacaagaaag ggatcttcac
     2041 tcgcgaccgc aaaccgaagt cggcggcttt tctgctgcaa aaacgctgga ctggcatgaa
     2101 cttcggtgaa aaaccgcagc agggaggcaa acaacccggg atccggtaac taactaaggt
     2161 acccagctct tctggtttgg tttggacctc tggtcctgca acttgaggta gtcaagatgc
     2221 ataataaata acggattgtg tccgtaatca cacgtggtgc gtacgataac gcatagtgtt
     2281 tttccctcca cttaaatcga agggttgtgt cttggatcgc gcgggtcaaa tgtatatggt
     2341 tcatatacat ccgcaggcac gtaataaagc gaggggttcg aatccccccg ttacccccgg
     2401 taggggccca tatatgcgga attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     2461 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     2521 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     2581 gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     2641 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     2701 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     2761 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2821 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2881 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2941 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3001 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     3061 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3121 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     3181 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3241 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3301 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3361 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     3421 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3481 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3541 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3601 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3661 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     3721 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3781 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3841 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3901 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3961 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4021 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     4081 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4141 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4201 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4261 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4321 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     4381 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4441 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4501 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4561 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4621 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4681 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4741 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4801 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4861 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4921 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     4981 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     5041 cccgactgga aagcgggcag tgagcg
//
