LOCUS pIX4.0-Luc 4804 bp ds-DNA circular SYN DEFINITION Control Vector Luc ACCESSION . VERSION . KEYWORDS pIX4.0-Luc; RTS; cell-free protein synthesis SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4804) AUTHORS biotechrabbit GmbH Volmerstr. 9, 12489 Berlin, Germany COMMENT Luc (firefly luciferase) expression vector with C-terminal 6xHis-tag for the control reaction. Ordering information: BR1401501 RTS 100 Insect Membrane Kit BR1501502 RTS 100 Insect Membrane Kit FEATURES Location/Qualifiers source 1..4804 /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" misc_feature 198..248 /label=5'-UTR /note="5'-polyhedrin-UTR" CDS 251..1927 /codon_start=1 /gene="luc" /product="firefly luciferase (Photinus pyralis) with C-Terminal 6xHis-tag" /label=Luc-6xHis /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAVWGHHHHHH" misc_feature 1970..1999 /label=polyA /note="polyadenylation site, adds 30 A to the mRNA" primer_bind complement(2050..2068) /label=T7-rev /note="3' sequencing primer" terminator 2064..2111 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" misc_feature 2114..2136 /label=Linearization site /note="Linearization site for production of run-off transcripts. blunt ends: DraI, EcoRV, SmaI, StuI, SwaI 5'-overhang: EcoRI, XmaI" rep_origin complement(2553..3141) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3312..4172) /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(4173..4277) /gene="bla" /label=AmpR promoter ORIGIN 1 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt 61 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg 121 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc ctaatacgac 181 tcactatagg gtctagataa gtattttact gttttcgtaa cagttttgta ataaaaaaac 241 ctataaatcc atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct 301 ggaagatgga accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc 361 tggaacaatt gcttttacag atgcacatat cgaggtggac atcacttacg ctgagtactt 421 cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag 481 aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt 541 tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag 601 tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt 661 gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt atcatggatt ctaaaacgga 721 ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa 781 tgaatacgat tttgtgccag agtccttcga tagggacaag acaattgcac tgatcatgaa 841 ctcctctgga tctactggtc tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt 901 gagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat 961 tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat 1021 atgtggattt cgagtcgtct taatgtatag atttgaagaa gagctgtttc tgaggagcct 1081 tcaggattac aagattcaaa gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa 1141 aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt ctggtggcgc 1201 tcccctctct aaggaagtcg gggaagcggt tgccaagagg ttccatctgc caggtatcag 1261 gcaaggatat gggctcactg agactacatc agctattctg attacacccg agggggatga 1321 taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga 1381 taccgggaaa acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat 1441 tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg 1501 gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca tcgttgaccg 1561 cctgaagtct ctgattaagt acaaaggcta tcaggtggct cccgctgaat tggaatccat 1621 cttgctccaa caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg acgatgacgc 1681 cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga 1741 gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt 1801 gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga 1861 gatcctcata aaggccaaga agggcggaaa gatcgccgtg tggggtcacc accatcacca 1921 tcattaataa ctaactaacc aagatctaaa taataagtaa ttaactagta aaaaaaaaaa 1981 aaaaaaaaaa aaaaaaaaag gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg 2041 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga 2101 ggggtttttt gatatcccgg gatttaaata ggcctgaatt cgtaatcatg gtcatagctg 2161 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 2221 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 2281 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 2341 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 2401 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 2461 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 2521 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 2581 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 2641 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 2701 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 2761 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 2821 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 2881 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 2941 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta 3001 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 3061 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 3121 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 3181 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 3241 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 3301 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 3361 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 3421 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 3481 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 3541 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 3601 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 3661 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 3721 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 3781 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 3841 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 3901 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 3961 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 4021 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 4081 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 4141 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 4201 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 4261 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt 4321 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt 4381 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt 4441 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg 4501 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg 4561 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca 4621 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag 4681 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag 4741 tcacgacgtt gtaaaacgac ggccagtgcc aagcttgcat gcaaggagat ggcgcccaac 4801 agtc //