LOCUS       pQE2-EF-Ts                5697 bp ds-DNA     circular SYN
DEFINITION  Control Vector EF-Ts
ACCESSION   .
VERSION     .
KEYWORDS    pQE2-EF-Ts; RTS; cell-free protein synthesis
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5697)
            Volmerstr. 9, 12489 Berlin, Germany
COMMENT     Ef-Ts (elongation factor Ts) expression vector with C-terminal 
            6xHis-tag for the control reaction.
            Ordering information: 
            BR1402401  RTS Linear Template Kit Plus
FEATURES             Location/Qualifiers
     source          1..5697
                     /organism="synthetic DNA construct"
                     /mol_type="circular plasmid"
     promoter        10..54
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    62..78
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             115..990
                     /codon_start=1
                     /gene="tsf"
                     /product="elongation factor Ts (Escherichia coli) with 
                     C-terminal 6xHis-tag"
                     /label=EF-Ts-6xHis
                     /translation="MAEITASLVKELRERTGAGMMDCKKALTEANGDIELAIENMRKSG
                     AIKAAKKAGNVAADGVIKTKIDGNYGIILEVNCQTDFVAKDAGFQAFADKVLDAAVAGK
                     ITDVEVLKAQFEEERVALVAKIGENINIRRVAALEGDVLGSYQHGARIGVLVAAKGADE
                     ELVKHIAMHVAASKPEFIKPEDVSAEVVEKEYQVQLDIAMQSGKPKEIAEKMVEGRMKK
                     FTGEVSLTGQPFVMEPSKTVGQLLKEHNAEVTGFIRFEVGEGIEKVETDFAAEVAAMSK
                     QSWGHHHHHH"
     terminator      1069..1116
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1154..1248
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      2016..2102
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2195..3277)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(3278..3355)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     misc_feature    3547..3687
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3873..4461)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4632..5492)
                     /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(5493..5597)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatggct
      121 gaaattaccg catccctggt aaaagagctg cgtgagcgta ctggcgcagg catgatggat
      181 tgcaaaaaag cactgactga agctaacggc gacatcgagc tggcaatcga aaacatgcgt
      241 aagtccggtg ctattaaagc agcgaaaaaa gcaggcaacg ttgctgctga cggcgtgatc
      301 aaaaccaaaa tcgacggcaa ctacggcatc attctggaag ttaactgcca gactgacttc
      361 gttgcaaaag acgctggttt ccaggcgttc gcagacaaag ttctggacgc agctgttgct
      421 ggcaaaatca ctgacgttga agttctgaaa gcacagttcg aagaagaacg tgttgcgctg
      481 gtagcgaaaa ttggtgaaaa catcaacatt cgccgcgttg ctgcgctgga aggcgacgtt
      541 ctgggttctt atcagcacgg tgcgcgtatc ggcgttctgg ttgctgctaa aggcgctgac
      601 gaagagctgg ttaaacacat cgctatgcac gttgctgcaa gcaagccaga attcatcaaa
      661 ccggaagacg tatccgctga agtggtagaa aaagaatacc aggtacagct ggatatcgcg
      721 atgcagtctg gtaagccgaa agaaatcgca gagaaaatgg ttgaaggccg catgaagaaa
      781 ttcaccggcg aagtttctct gaccggtcag ccgttcgtta tggaaccaag caaaactgtt
      841 ggtcagctgc tgaaagagca taacgctgaa gtgactggct tcatccgctt cgaagtgggt
      901 gaaggcatcg agaaagttga gactgacttt gcagcagaag ttgctgcgat gtccaagcag
      961 tcttggggtc accaccatca ccatcattaa taactaacta accaagatcc ggctgctaac
     1021 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     1081 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaagatc gaaagcttaa
     1141 ttagctgagc ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat
     1201 ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagctagctt
     1261 ggcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac
     1321 cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca
     1381 atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa
     1441 aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca
     1501 tccggaattt cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc
     1561 ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca
     1621 cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa
     1681 cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg
     1741 ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt
     1801 tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca
     1861 ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca
     1921 gtactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta
     1981 aacgcctggg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag gctcagtcga
     2041 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     2101 atccgccctc tagattacgt gcagtcgatg ataagctgtc aaacatgaga attgtgccta
     2161 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2221 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2281 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     2341 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     2401 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     2461 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     2521 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     2581 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     2641 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     2701 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     2761 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     2821 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     2881 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     2941 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     3001 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     3061 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     3121 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     3181 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     3241 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     3301 cttccgggcg ctatcatgcc ataccgcgaa aggttttgca ccattcgatg gtgtcggaat
     3361 ttcgggcagc gttgggtcct ggccacgggt gcgcatgatc tagagctgcc tcgcgcgttt
     3421 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     3481 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     3541 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
     3601 gcggcatcag agcagattgt actgagagtg caccacatgc ggtgtgaaat accgcacaga
     3661 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     3721 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3781 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3841 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3901 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3961 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4021 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4081 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4141 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4201 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4261 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     4321 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4381 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4441 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4501 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4561 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4621 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4681 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4741 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4801 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4861 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4921 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4981 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5041 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5101 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5161 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5221 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5281 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5341 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5401 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5461 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5521 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5581 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     5641 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcac
//
