LOCUS       40924_43069        5404 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTEV5, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5404)
  AUTHORS   Rocco CJ, Dennison KL, Klenchin VA, Rayment I, Escalante-Semerena 
            JC.
  TITLE     Construction and use of new cloning vectors for the rapid isolation 
            of recombinant proteins from Escherichia coli
  JOURNAL   Plasmid 59 (3), 231-237 (2008)
  PUBMED    18295882
REFERENCE   2  (bases 1 to 5404)
  AUTHORS   Rocco CJ, Dennison KL, Klenchin VA, Rayment I, Escalante-Semerena 
            JC.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-DEC-2007) Bacteriology, University of Wisconsin - 
            Madison, 1550 Linden Dr., Madison, WI 53706, USA
REFERENCE   3  (bases 1 to 5404)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5404)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid"; 
            date: "2008"; volume: "59"; issue: "3"; pages: "231-237"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-DEC-2007) Bacteriology, University of Wisconsin - Madison, 1550 
            Linden Dr., Madison, WI 53706, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             18..35
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             57..77
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     terminator      137..184
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      221..676
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        703..807
                     /label=AmpR promoter
     CDS             808..1665
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1839..2421
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2607..2749)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2854..3042)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(3817..3838)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3854..4933)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4934..5011)
                     /label=lacI promoter
     promoter        5320..5338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5339..5363
                     /label=lac operator
                     /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             5378..5400
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 catatatgtc gtactaccat caccatcacc atcacgatta cgatatccca actagtgaaa
       61 acctgtattt tcagggcgct agccatatgg ccatggaatt ctcgagctcc cgggatccgc
      121 ggccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
      181 tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca
      241 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
      301 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
      361 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
      421 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
      481 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
      541 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      601 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      661 ttaacgttta caatttcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
      721 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
      781 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
      841 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
      901 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
      961 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     1021 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     1081 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     1141 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     1201 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     1261 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     1321 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     1381 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     1441 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     1501 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     1561 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     1621 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     1681 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     1741 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1801 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1861 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1921 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1981 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2041 acatacctcg ctctgctaat cctgttacca gctgccagtg gcgataagtc gtgtcttacc
     2101 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2161 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2221 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2281 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2341 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2401 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2461 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     2521 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     2581 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     2641 ggtatttcac accgcatata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     2701 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     2761 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     2821 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     2881 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     2941 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     3001 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg
     3061 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
     3121 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
     3181 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac
     3241 agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat
     3301 ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat
     3361 tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg
     3421 tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa
     3481 cgacaggagc acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt
     3541 ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat
     3601 tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc
     3661 gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt
     3721 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     3781 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     3841 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3901 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     3961 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     4021 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     4081 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac
     4141 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     4201 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     4261 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     4321 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     4381 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     4441 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     4501 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     4561 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     4621 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     4681 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     4741 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     4801 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     4861 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     4921 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     4981 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     5041 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     5101 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     5161 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     5221 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     5281 cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg
     5341 aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga
     5401 tata
//
