LOCUS       40924_4969        4366 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pAT223, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4366)
  AUTHORS   Amstutz P, Binz HK, Parizek P, Stumpp MT, Kohl A, Grutter MG, Forrer
            P, Pluckthun A.
  TITLE     Intracellular kinase inhibitors selected from combinatorial 
            libraries of designed ankyrin repeat proteins
  JOURNAL   J. Biol. Chem. 280 (26), 24715-24722 (2005)
  PUBMED    15851475
REFERENCE   2  (bases 1 to 4366)
  AUTHORS   Stumpp MT, Forrer P, Binz HK, Amstutz P, Zahnd C, Pluckthun A.
  TITLE     Designed ankyrin repeat proteins binding maltose binding protein
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 4366)
  AUTHORS   Stumpp MT, Forrer P, Binz HK, Amstutz P, Zahnd C, Pluckthun A.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JUN-2003) Biochemistry, University of Zurich, 
            Winterthurerstrasse 190, Zurich 8057, Switzerland
REFERENCE   4  (bases 1 to 4366)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 4366)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Biol. 
            Chem."; date: "2005"; volume: "280"; issue: "26"; pages: 
            "24715-24722"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (20-JUN-2003) Biochemistry, University of Zurich, 
            Winterthurerstrasse 190, Zurich 8057, Switzerland"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4366
                     /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..456
                     /label=bacteriophage lambda protein D
                     /note="bacteriophage lambda protein D"
     CDS             457..474
                     /label=6xHis
                     /note="6xHis affinity tag"
     misc_feature    475..519
                     /label=multiple cloning site
                     /note="multiple cloning site"
     terminator      529..623
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      730..816
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(879..1958)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(1959..2036)
                     /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    2216..2356
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2542..3130)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3304..4161)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4162..4266)
                     /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 agcggtcatc accatcacca tcacggatcc
      481 ggtggtaccc cgggtcgacc tgcagccaag cttaattagc tgagcttgga ctcctgttga
      541 tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc
      601 gggcgttttt tattggtgag aatccaagct agcagtactg cgatgagtgg cagggcgggg
      661 cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc tggggtaatg actctctagt
      721 ttgaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
      781 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgctctagag atttccctcg
      841 acaattcgcg ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
      901 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
      961 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
     1021 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
     1081 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
     1141 tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
     1201 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
     1261 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
     1321 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
     1381 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
     1441 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
     1501 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
     1561 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
     1621 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
     1681 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
     1741 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
     1801 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
     1861 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
     1921 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     1981 tcttccgggc gctatcatgc cataccgcga aaggttttgc acctttcgat ggtgtcaacg
     2041 taaatgcatg ccgcttcgcc ttccctagct agagctgcct cgcgcgtttc ggtgatgacg
     2101 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     2161 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
     2221 ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga
     2281 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     2341 aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtctg
     2401 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2461 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2521 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2581 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2641 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2701 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     2761 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2821 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2881 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2941 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     3001 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3061 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3121 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3181 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3241 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3301 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3361 agctgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3421 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3481 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3541 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3601 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3661 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3721 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3781 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3841 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3901 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3961 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4021 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4081 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4141 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4201 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4261 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4321 attaacctat aaaaataggc gtatcacgag gccctttcgt cttcac
//
