LOCUS       40924_43049        6610 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTEV19, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6610)
  AUTHORS   VanDrisse CM, Escalante-Semerena JC.
  TITLE     New high-cloning-efficiency vectors for complementation studies and 
            recombinant protein overproduction in Escherichia coli and 
            Salmonella enterica
  JOURNAL   Plasmid (2016) In press
  PUBMED    27234933
REFERENCE   2  (bases 1 to 6610)
  AUTHORS   VanDrisse CM, Escalante-Semerena JC.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-MAR-2016) Microbiology, University of Georgia, 120 
            Cedar St., Athens, GA 30602, USA
REFERENCE   3  (bases 1 to 6610)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6610)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid 
            (2016) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-MAR-2016) Microbiology, University of Georgia, 120 Cedar St., 
            Athens, GA 30602, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             56..73
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      140..187
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      224..679
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        706..810
                     /label=AmpR promoter
     CDS             811..1668
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1842..2430
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2616..2758)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2863..3051)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(3826..3847)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3863..4942)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4943..5020)
                     /label=lacI promoter
     promoter        5329..5347
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5348..5372
                     /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             5387..5409
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5426..5443
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             5450..6547
                     /codon_start=1
                     /label=MBP
                     /note="maltose binding protein from E. coli"
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     CDS             join(6593..6610,1..3)
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
ORIGIN
        1 ggtttcagaa gagcgctctt cctaacctgc aggcaagctt gcggccgcac tcgagcacca
       61 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
      121 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
      181 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
      241 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
      301 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
      361 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
      421 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
      481 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
      541 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
      601 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
      661 atattaacgt ttacaatttc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
      721 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
      781 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
      841 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
      901 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
      961 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1021 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     1081 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1141 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     1201 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     1261 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     1321 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     1381 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     1441 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     1501 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     1561 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     1621 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     1681 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     1741 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     1801 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1861 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1921 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1981 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2041 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2101 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2161 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2221 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2281 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2341 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2401 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2461 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     2521 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     2581 cgcagcgagt cagtgagcga ggaagcggaw gwgcgcctga tgcggtattt tctccttacg
     2641 catctgtgcg gtatttcaca ccgcatatat ggtgcactct cagtacaatc tgctctgatg
     2701 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     2761 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     2821 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     2881 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     2941 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     3001 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     3061 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     3121 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     3181 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     3241 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     3301 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     3361 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     3421 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     3481 ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggcgataat
     3541 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     3601 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     3661 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     3721 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     3781 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     3841 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3901 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     3961 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     4021 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     4081 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     4141 tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     4201 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     4261 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     4321 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     4381 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     4441 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     4501 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     4561 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     4621 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     4681 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     4741 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     4801 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     4861 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     4921 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     4981 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     5041 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
     5101 gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
     5161 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
     5221 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     5281 gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaatta atacgactca
     5341 ctatagggga attgtgagcg gataacaatt cccctctaga aataattttg tttaacttta
     5401 agaaggagat atacatatgg gcagccatca ccatcaccat cactccggta aaatcgaaga
     5461 aggtaaactg gtaatctgga ttaacggcga taaaggctat aacggtctcg ctgaagtcgg
     5521 taagaaattc gagaaagata ccggaattaa agtcaccgtt gagcatccgg ataaactgga
     5581 agagaaattc ccacaggttg cggcaactgg cgatggccct gacattatct tctgggcaca
     5641 cgaccgcttt ggtggctacg ctcaatctgg cctgttggct gaaatcaccc cggacaaagc
     5701 gttccaggac aagctgtatc cgtttacctg ggatgccgta cgttacaacg gcaagctgat
     5761 tgcttacccg atcgctgttg aagcgttatc gctgatttat aacaaagatc tgctgccgaa
     5821 cccgccaaaa acctgggaag agatcccggc gctggataaa gaactgaaag cgaaaggtaa
     5881 gagcgcgctg atgttcaacc tgcaagaacc gtacttcacc tggccgctga ttgctgctga
     5941 cgggggttat gcgttcaagt atgaaaacgg caagtacgac attaaagacg tgggcgtgga
     6001 taacgctggc gcgaaagcgg gtctgacctt cctggttgac ctgattaaaa acaaacacat
     6061 gaatgcagac accgattact ccatcgcaga agctgccttt aataaaggcg aaacagcgat
     6121 gaccatcaac ggcccgtggg catggtccaa catcgacacc agcaaagtga attatggtgt
     6181 aacggtactg ccgaccttca agggtcaacc atccaaaccg ttcgttggcg tgctgagcgc
     6241 aggtattaac gccgccagtc cgaacaaaga gctggcaaaa gagttcctcg aaaactatct
     6301 gctgactgat gaaggtctgg aagcggttaa taaagacaaa ccgctgggtg ccgtagcgct
     6361 gaagtcttac gaggaagagt tggcgaaaga tccacgtatt gccgccacta tggaaaacgc
     6421 ccagaaaggt gaaatcatgc cgaacatccc gcagatgtcc gctttctggt atgccgtgcg
     6481 tactgcggtg atcaacgccg ccagcggtcg tcagactgtc gatgaagccc tgaaagacgc
     6541 gcagactaat tcgagctcga gcggaaccgc ctcgggcggt gcaaccacgt cagagaatct
     6601 ctacttccaa
//
