LOCUS       40924_33187        7602 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNH-TrxT, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7602)
  AUTHORS   Cooper CDO., Gileadi O.
  TITLE     
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7602)
  AUTHORS   Cooper CDO., Gileadi O.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-DEC-2009) Structural Genomics Consortium, University 
            of Oxford, ORCRB, Old Road Campus, off Roosevelt Drive, Oxford, 
            Oxfordshire OX3 7DQ, United Kingdom
REFERENCE   3  (bases 1 to 7602)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7602)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-DEC-2009) Structural Genomics Consortium, University of Oxford, 
            ORCRB, Old Road Campus, off Roosevelt Drive, Oxford, Oxfordshire OX3
            7DQ, United Kingdom"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             12..34
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             45..62
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             72..398
                     /label=TrxA
                     /note="E. coli thioredoxin"
     CDS             405..425
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     promoter        441..886
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             887..2305
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     misc_feature    2354..2368
                     /note="3'-LIC sequence; add complement to downstream PCR
                     primer"
     CDS             2416..2433
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      2500..2547
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2584..3039
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(3135..3947)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      4069..4657
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4843..4985)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(5090..5278)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(6053..6074)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(6090..7169)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(7170..7247)
                     /label=lacI promoter
     promoter        7556..7574
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7575..7599
                     /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)."
ORIGIN
        1 ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgcaccat catcatcatc
       61 attcttctgg tatgagcgat aaaattattc acctgactga cgacagtttt gacacggatg
      121 tactcaaagc ggacggggcg atcctcgtcg atttctgggc agagtggtgc ggtccgtgca
      181 aaatgatcgc cccgattctg gatgaaatcg ctgacgaata tcagggcaaa ctgaccgttg
      241 caaaactgaa catcgatcaa aaccctggca ctgcgccgaa atatggcatc cgtggtatcc
      301 cgactctgct gctgttcaaa aacggtgaag tggcggcaac caaagtgggc gcactgtcta
      361 aaggtcagtt gaaagagttc ctcgacgcta acctggccgg taccgagaac ttgtacttcc
      421 aatccatgga gaccgacgtc cacatatacc tgccgttcac tattatttag tgaaatgaga
      481 tattatgata ttttctgaat tgtgattaaa aaggcaactt tatgcccatg caacagaaac
      541 tataaaaaat acagagaatg aaaagaaaca gatagatttt ttagttcttt aggcccgtag
      601 tctgcaaatc cttttatgat tttctatcaa acaaaagagg aaaatagacc agttgcaatc
      661 caaacgagag tctaatagaa tgaggtcgaa aagtaaatcg cgcgggtttg ttactgataa
      721 agcaggcaag acctaaaatg tgtaaagggc aaagtgtata ctttggcgtc accccttaca
      781 tattttaggt ctttttttat tgtgcgtaac taacttgcca tcttcaaaca ggagggctgg
      841 aagaagcaga ccgctaacac agtacataaa aaaggagaca tgaacgatga acatcaaaaa
      901 gtttgcaaaa caagcaacag tattaacctt tactaccgca ctgctggcag gaggcgcaac
      961 tcaagcgttt gcgaaagaaa cgaaccaaaa gccatataag gaaacatacg gcatttccca
     1021 tattacacgc catgatatgc tgcaaatccc tgaacagcaa aaaaatgaaa aatataaagt
     1081 tcctgagttc gattcgtcca caattaaaaa tatctcttct gcaaaaggcc tggacgtttg
     1141 ggacagctgg ccattacaaa acactgacgg cactgtcgca aactatcacg gctaccacat
     1201 cgtctttgca ttagccggag atcctaaaaa tgcggatgac acatcgattt acatgttcta
     1261 tcaaaaagtc ggcgaaactt ctattgacag ctggaaaaac gctggccgcg tctttaaaga
     1321 cagcgacaaa ttcgatgcaa atgattctat cctaaaagac caaacacaag aatggtcagg
     1381 ttcagccaca tttacatctg acggaaaaat ccgtttattc tacactgatt tctccggtaa
     1441 acattacggc aaacaaacac tgacaactgc acaagttaac gtatcagcat cagacagctc
     1501 tttgaacatc aacggtgtag aggattataa atcaatcttt gacggtgacg gaaaaacgta
     1561 tcaaaatgta cagcagttca tcgatgaagg caactacagc tcaggcgaca accatacgct
     1621 gagagatcct cactacgtag aagataaagg ccacaaatac ttagtatttg aagcaaacac
     1681 tggaactgaa gatggctacc aaggcgaaga atctttattt aacaaagcat actatggcaa
     1741 aagcacatca ttcttccgtc aagaaagtca aaaacttctg caaagcgata aaaaacgcac
     1801 ggctgagtta gcaaacggcg ctctcggtat gattgagcta aacgatgatt acacactgaa
     1861 aaaagtgatg aaaccgctga ttgcatctaa cacagtaaca gatgaaattg aacgcgcgaa
     1921 cgtctttaaa atgaacggca aatggtacct gttcactgac tcccgcggat caaaaatgac
     1981 gattgacggc attacgtcta acgatattta catgcttggt tatgtttcta attctttaac
     2041 tggcccatac aagccgctga acaaaactgg ccttgtgtta aaaatggatc ttgatcctaa
     2101 cgatgtaacc tttacttact cacacttcgc tgtacctcaa gcgaaaggaa acaatgtcgt
     2161 gattacaagc tatatgacaa acagaggatt ctacgcagac aaacaatcaa cgtttgcgcc
     2221 tagcttcctg ctgaacatca aaggcaagaa aacatctgtt gtcaaagaca gcatccttga
     2281 acaaggacaa ttaacagtta acaaataaaa acgcaaaaga aaatgccgat atcctattgg
     2341 cattgacggt ctccagtaaa ggtggatacg gatccgaatt cgagctccgt cgacaagctt
     2401 gcggccgcac tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga
     2461 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     2521 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat
     2581 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2641 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2701 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2761 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     2821 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2881 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     2941 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3001 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc aggtggcact tttcggggaa
     3061 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3121 tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     3181 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     3241 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     3301 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     3361 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt
     3421 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     3481 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa
     3541 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     3601 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt
     3661 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     3721 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     3781 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct
     3841 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     3901 tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac accccttgta
     3961 ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt aacgtgagtt
     4021 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4081 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4141 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4201 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4261 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4321 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4381 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4441 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4501 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4561 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4621 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4681 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4741 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     4801 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     4861 ccttacgcat ctgtgcggta tttcacaccg catatatggt gcactctcag tacaatctgc
     4921 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     4981 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     5041 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     5101 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     5161 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     5221 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     5281 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     5341 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     5401 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     5461 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     5521 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     5581 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     5641 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     5701 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg
     5761 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
     5821 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc
     5881 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca
     5941 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
     6001 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
     6061 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6121 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
     6181 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
     6241 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
     6301 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
     6361 accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
     6421 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
     6481 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
     6541 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
     6601 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
     6661 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
     6721 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
     6781 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
     6841 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
     6901 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
     6961 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
     7021 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
     7081 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
     7141 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
     7201 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg
     7261 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
     7321 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
     7381 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
     7441 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     7501 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata
     7561 cgactcacta taggggaatt gtgagcggat aacaattccc ct
//
