LOCUS       40924_43059        5402 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTEV3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5402)
  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 5402)
  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 5402)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5402)
  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..5402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             16..33
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             55..75
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     terminator      135..182
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      219..674
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        701..805
                     /label=AmpR promoter
     CDS             806..1663
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1837..2419
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2605..2747)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2852..3040)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(3815..3836)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3852..4931)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4932..5009)
                     /label=lacI promoter
     promoter        5318..5336
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5337..5361
                     /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             5376..5398
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 catatgtcgt actaccatca ccatcaccat cacgattacg atatcccaac tagtgaaaac
       61 ctgtattttc agggcgctag cgcgcccatg ggcgaattct cgagctcccg ggatccgcgg
      121 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
      181 tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt
      241 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
      301 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
      361 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
      421 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
      481 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
      541 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
      601 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
      661 aacgtttaca atttcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
      721 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
      781 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
      841 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
      901 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
      961 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     1021 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     1081 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     1141 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     1201 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     1261 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     1321 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     1381 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     1441 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     1501 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     1561 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     1621 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     1681 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     1741 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     1801 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1861 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1921 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     1981 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2041 atacctcgct ctgctaatcc tgttaccagc tgccagtggc gataagtcgt gtcttaccgg
     2101 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2161 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2221 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2281 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2341 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2401 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2461 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2521 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     2581 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2641 tatttcacac cgcatatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     2701 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     2761 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     2821 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     2881 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     2941 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     3001 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
     3061 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
     3121 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
     3181 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
     3241 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
     3301 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
     3361 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
     3421 tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg
     3481 acaggagcac gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct
     3541 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     3601 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     3661 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     3721 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     3781 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
     3841 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3901 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
     3961 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
     4021 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
     4081 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca
     4141 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     4201 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
     4261 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
     4321 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
     4381 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
     4441 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
     4501 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
     4561 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
     4621 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
     4681 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
     4741 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     4801 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     4861 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     4921 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     4981 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
     5041 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
     5101 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     5161 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     5221 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     5281 tccggcgtag aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa
     5341 ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata
     5401 ta
//
