LOCUS       40924_45948        4985 bp DNA     circular SYN 18-DEC-2018
DEFINITION  T7 expression vector pViet, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4985)
  AUTHORS   Honag TT, Stern RJ, McNeil MR, Schweizer HP.
  TITLE     Construction and use of low-copy number T7 expression vectors for 
            purification of problem proteins: affinity purification of 
            Mycobacterium tuberculosis RmlD and Pseudomonas aeruginosa LasI and 
            RhlI proteins, and functional analysis of purified RhlI
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4985)
  AUTHORS   Hoang TT, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-APR-1999) Microbiology, Colorado State University, 
            Fort Collins, CO 80523, USA
REFERENCE   3  (bases 1 to 4985)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4985)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-APR-1999) Microbiology, Colorado State University, Fort Collins,
            CO 80523, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(147..1004)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1005..1109)
                     /label=AmpR promoter
     promoter        1222..1250
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     terminator      complement(1425..1472)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1548..1565)
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     CDS             complement(1575..1592)
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     RBS             complement(1611..1633)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(1648..1672)
                     /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)."
     promoter        complement(1673..1691)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2004..2081
                     /label=lacI promoter
     CDS             2082..3161
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    3177..3198
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      3663..3885
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             3933..4880
                     /codon_start=1
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
       61 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
      121 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
      181 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
      241 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
      301 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
      361 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
      421 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
      481 tcgtcgtttg gtatggcttc attcagctcc ggttcccagc gatcaaggcg agttacatga
      541 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
      601 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
      661 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
      721 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
      781 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
      841 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
      901 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
      961 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     1021 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     1081 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     1141 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     1201 cgtcttcaag aattctcatg tttgacagct tatcatcgat aagctttaat gcggtagttt
     1261 atcacagtta aattgctaac gcagtcaggc accgtgtatg aaatctaaca atgcgctcat
     1321 cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata ggcttggtta tgccggtact
     1381 gccgggcctc ttgcgggata tccggatata gttcctcctt tcagcaaaaa acccctcaag
     1441 acccgtttag aggccccaag gggttatgct agttattgct cagcggtggc agcagccaac
     1501 tcagcttcct ttcgggcttt gttagcagcc ggatcctcga gcatatggct gccgcgcggc
     1561 accaggccgc tgctgtgatg atgatgatga tggctgctgc ccatggtata tctccttctt
     1621 aaagttaaac aaaattattt ctagagggga attgttatcc gctcacaatt cccctatagt
     1681 gagtcgtatt aatttcgcgg gatcgagatc gatctcgatc ctctacgccg gacgcatcgt
     1741 ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga
     1801 tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt
     1861 ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcatgcac cattccttgc
     1921 ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc aggagtcgca
     1981 taagggagag cgtcgagatc ccggacacca tcgaatggcg caaaaccttt cgcggtatgg
     2041 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
     2101 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     2161 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
     2221 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
     2281 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
     2341 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
     2401 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
     2461 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat
     2521 ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat gaagacggta
     2581 cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg
     2641 gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc
     2701 gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc
     2761 aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg
     2821 atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg
     2881 atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa
     2941 ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac
     3001 tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa
     3061 aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     3121 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     3181 gtaagttagc tcactcatta ggcaccggga tctcgaccga tgcccttgag agccttcaac
     3241 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
     3301 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag
     3361 gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg
     3421 cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag
     3481 gccattatcg ccggcatggc ggccgacgcg ctgggctacg tcttgctggc gttcgattca
     3541 gcgatttgcc cgagcttgcg agggtgctac ttaagccttt agggttttaa ggtctgtttt
     3601 gtagaggagc aaacagcgtt tgcgacatcc ttttgtaata ctgcggaact gactaaagta
     3661 gtgagttata cacagggctg ggatctattc tttttatctt tttttattct ttctttattc
     3721 tataaattat aaccacttga atataaacaa aaaaaacaca caaaggtcta gcggaattta
     3781 cagagggtct agcagaattt acaagttttc cagcaaaggt ctagcagaat ttacagatac
     3841 ccacaactca aaggaaaagg actagtaatt atcattgact agcccatctc aattggtata
     3901 gtgattaaaa tcacctagac caattgagat gtatgtctga attagttgtt ttcaaagcaa
     3961 atgaactagc gattagtcgc tatgacttaa cggagcatga aaccaagcta attttatgct
     4021 gtgtggcact actcaacccc acgattgaaa accctacaag gaaagaacgg acggtatcgt
     4081 tcacttataa ccaatacgct cagatgatga acatcagtag ggaaaatgct tatggtgtat
     4141 tagctaaagc aaccagagag ctgatgacga gaactgtgga aatcaggaat cctttggtta
     4201 aaggctttga gattttccag tggacaaact atgccaagtt ctcaagcgaa aaattagaat
     4261 tagtttttag tgaagagata ttgccttatc ttttccagtt aaaaaaattc ataaaatata
     4321 atctggaaca tgttaagtct tttgaaaaca aatactctat gaggatttat gagtggttat
     4381 taaaagaact aacacaaaag aaaactcaca aggcaaatat agagattagc cttgatgaat
     4441 ttaagttcat gttaatgctt gaaaataact accatgagtt taaaaggctt aaccaatggg
     4501 ttttgaaacc aataagtaaa gatttaaaca cttacagcaa tatgaaattg gtggttgata
     4561 agcgaggccg cccgactgat acgttgattt tccaagttga actagataga caaatggatc
     4621 tcgtaaccga acttgagaac aaccagataa aaatgaatgg tgacaaaata ccaacaacca
     4681 ttacatcaga ttcctaccta cataacggac taagaaaaac actacacgat gctttaactg
     4741 caaaaattca gctcaccagt tttgaggcaa aatttttgag tgacatgcaa agtaagtatg
     4801 atctcaatgg ttcgttctca tggctcacgc aaaaacaacg aaccacacta gagaacatac
     4861 tggctaaata cggaaggatc tgaggttctt atggctcttg tatctatcag tgaagcatca
     4921 agactaacaa acaaaagtag aacaactgtt caccgttaca tatcaaaggg aaaactgtcc
     4981 ataga
//
