LOCUS       40924_43054        5361 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTEV20, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5361)
  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 5361)
  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 5361)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5361)
  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..5361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             34..54
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             55..72
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      94..141
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      178..633
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        660..764
                     /label=AmpR promoter
     CDS             765..1622
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1796..2378
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2564..2706)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2811..2999)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(3774..3795)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3811..4890)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4891..4968)
                     /label=lacI promoter
     promoter        5277..5295
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5296..5320
                     /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             5335..5357
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 catatatgtc gtactacaga agagcgctct tcagaaaacc tgtattttca gggccatcac
       61 catcaccatc actgagcggc cgctgagcaa taactagcat aaccccttgg ggcctctaaa
      121 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg
      181 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
      241 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
      301 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
      361 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
      421 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
      481 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
      541 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
      601 cgaattttaa caaaatatta acgtttacaa tttcaggtgg cacttttcgg ggaaatgtgc
      661 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
      721 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
      781 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
      841 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
      901 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
      961 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     1021 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     1081 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     1141 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     1201 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     1261 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     1321 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     1381 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     1441 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     1501 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     1561 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     1621 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     1681 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     1741 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1801 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1861 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1921 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     1981 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagct gccagtggcg
     2041 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2101 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2161 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2221 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2281 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2341 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2401 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2461 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     2521 cgaccgagcg cagcgagtca gtgagcgagg aagcggawgw gcgcctgatg cggtattttc
     2581 tccttacgca tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg
     2641 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     2701 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     2761 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     2821 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     2881 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     2941 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     3001 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     3061 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
     3121 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
     3181 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     3241 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
     3301 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     3361 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
     3421 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg
     3481 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
     3541 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
     3601 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
     3661 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
     3721 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     3781 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3841 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3901 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     3961 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     4021 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     4081 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     4141 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     4201 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     4261 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     4321 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     4381 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     4441 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     4501 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     4561 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     4621 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     4681 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     4741 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     4801 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     4861 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     4921 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     4981 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     5041 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     5101 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     5161 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     5221 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat
     5281 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt
     5341 taactttaag aaggagatat a
//
