LOCUS       40924_4964        4361 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pAT222, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4361)
  AUTHORS   Binz HK, Amstutz P, Kohl A, Stumpp MT, Briand C, Forrer P, Grutter 
            MG, Pluckthun A.
  TITLE     High-affinity binders selected from designed ankyrin repeat protein 
            libraries
  JOURNAL   Nat. Biotechnol. 22 (5), 575-582 (2004)
  PUBMED    15097997
REFERENCE   2  (bases 1 to 4361)
  AUTHORS   Stumpp MT, Forrer P, Binz KH, Amstutz P, Zahnd C, Pluckthun A.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JUN-2003) Biochemistry, University of Zurich, 
            Winterthurerstrasse 190, Zurich 8057, Switzerland
REFERENCE   3  (bases 1 to 4361)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4361)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Biotechnol."; date: "2004"; volume: "22"; issue: "5"; pages: 
            "575-582"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (20-JUN-2003) Biochemistry, University of Zurich, 
            Winterthurerstrasse 190, Zurich 8057, Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..54
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA
                     polymerase, with embedded lac operator"
     protein_bind    62..78
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             121..165
                     /label=AviTag(TM)
                     /note="peptide tag that allows for enzymatic biotinylation"
     misc_feature    172..447
                     /note="truncated N-terminus bacteriophage lambda protein D"
     misc_feature    448..489
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             490..507
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      524..618
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      725..811
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(874..1953)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(1954..2031)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     misc_feature    2211..2351
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2537..3125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3299..4156)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4157..4261)
                     /label=AmpR promoter
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatggct
      121 ggtctgaacg atatcttcga agctcagaaa atcgaatggc acgaaggttc catgggcacc
      181 gcaaccgcgc ccggcggatt gagtgcgaaa gcgcctgcaa tgaccccgct gatgctggac
      241 acctccagcc gtaagctggt tgcgtgggat ggcaccaccg acggtgctgc cgttggcatt
      301 cttgcggttg ctgctgacca gaccagcacc acgctgacgt tctacaagtc cggcacgttc
      361 cgttatgagg atgtgctctg gccggaggct gccagcgacg agacgaaaaa acggaccgcg
      421 tttgccggaa cggcaatcag catcgttgga tccggtggta ccccgggtcg acctgcagcc
      481 caagcttctc atcaccatca ccatcactaa tgagctgagc ttggactcct gttgatagat
      541 ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg
      601 ttttttattg gtgagaatcc aagctagcag tactgcgatg agtggcaggg cggggcgtaa
      661 tttttttaag gcagttattg gtgcccttaa acgcctgggg taatgactct ctagtttgag
      721 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
      781 gtcggtgaac gctctcctga gtaggacaaa tccgccgctc tagagatttc cctcgacaat
      841 tcgcgctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
      901 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
      961 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     1021 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     1081 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     1141 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     1201 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     1261 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     1321 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     1381 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     1441 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     1501 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     1561 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     1621 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     1681 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     1741 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     1801 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     1861 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     1921 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     1981 cgggcgctat catgccatac cgcgaaaggt tttgcacctt tcgatggtgt caacgtaaat
     2041 gcatgccgct tcgccttccc tagctagagc tgcctcgcgc gtttcggtga tgacggtgaa
     2101 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     2161 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
     2221 acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga
     2281 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     2341 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtctgtcggc
     2401 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2461 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2521 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2581 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2641 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2701 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     2761 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2821 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2881 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2941 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     3001 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3061 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3121 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3181 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3241 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3301 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagctg
     3361 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3421 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3481 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3541 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3601 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3661 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3721 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3781 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3841 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3901 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3961 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4021 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4081 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4141 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4201 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4261 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     4321 cctataaaaa taggcgtatc acgaggccct ttcgtcttca c
//
