LOCUS       40924_4974        5189 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pAT224, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5189)
  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 5189)
  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 5189)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5189)
  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..5189
                     /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"
     CDS             178..1275
                     /label=MBP
                     /note="maltose binding protein from E. coli"
     misc_feature    1276..1317
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             1318..1335
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      1352..1446
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      1553..1639
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(1702..2781)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(2782..2859)
                     /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    3039..3179
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3365..3953)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4127..4984)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4985..5089)
                     /label=AmpR promoter
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatggct
      121 ggtctgaacg atatcttcga agctcagaaa atcgaatggc acgaaggttc catggggaaa
      181 actgaagaag gtaaactggt aatctggatt aacggcgata aaggctataa cggtctcgct
      241 gaagtcggta agaaattcga gaaagatacc ggaattaaag tcaccgttga gcatccggat
      301 aaactggaag agaaattccc acaggttgcg gcaactggcg atggccctga cattatcttc
      361 tgggcacacg accgctttgg tggctacgct caatctggcc tgttggctga aatcaccccg
      421 gacaaagcgt tccaggacaa gctgtatccg tttacctggg atgccgtacg ttacaacggc
      481 aagctgattg cttacccgat cgctgttgaa gcgttatcgc tgatttataa caaagatctg
      541 ctgccgaacc cgccaaaaac ctgggaagag atcccggcgc tggataaaga actgaaagcg
      601 aaaggtaaga gcgcgctgat gttcaacctg caagaaccgt acttcacctg gccgctgatt
      661 gctgctgacg ggggttatgc gttcaagtat gaaaacggca agtacgacat taaagacgtg
      721 ggcgtggata acgctggcgc gaaagcgggt ctgaccttcc tggttgacct gattaaaaac
      781 aaacacatga atgcagacac cgattactcc atcgcagaag ctgcctttaa taaaggcgaa
      841 acagcgatga ccatcaacgg cccgtgggca tggtccaaca tcgacaccag caaagtgaat
      901 tatggtgtaa cggtactgcc gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg
      961 ctgagcgcag gtattaacgc cgccagtccg aacaaagagc tggcaaaaga gttcctcgaa
     1021 aactatctgc tgactgatga aggtctggaa gcggttaata aagacaaacc gctgggtgcc
     1081 gtagcgctga agtcttacga ggaagagttg gcgaaagatc cacgtattgc cgccactatg
     1141 gaaaacgccc agaaaggtga aatcatgccg aacatcccgc agatgtccgc tttctggtat
     1201 gccgtgcgta ctgcggtgat caacgccgcc agcggtcgtc agactgtcga tgaagccctg
     1261 aaagacgcgc agactggatc cggtggtacc ccgggtcgac ctgcagccca agcttctcat
     1321 caccatcacc atcactaatg agctgagctt ggactcctgt tgatagatcc agtaatgacc
     1381 tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
     1441 gagaatccaa gctagcagta ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc
     1501 agttattggt gcccttaaac gcctggggta atgactctct agtttgaggc atcaaataaa
     1561 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     1621 tctcctgagt aggacaaatc cgccgctcta gagatttccc tcgacaattc gcgctaactt
     1681 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     1741 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     1801 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
     1861 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     1921 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
     1981 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     2041 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     2101 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     2161 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
     2221 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     2281 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     2341 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     2401 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     2461 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     2521 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     2581 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     2641 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     2701 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     2761 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     2821 tgccataccg cgaaaggttt tgcacctttc gatggtgtca acgtaaatgc atgccgcttc
     2881 gccttcccta gctagagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     2941 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     3001 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     3061 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
     3121 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     3181 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt ctgtcggctg cggcgagcgg
     3241 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3301 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3361 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3421 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3481 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3541 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     3601 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3661 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3721 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3781 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     3841 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3901 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3961 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4021 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4081 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4141 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagctgcc tgactccccg
     4201 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4261 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4321 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4381 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4441 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4501 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4561 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4621 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4681 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4741 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4801 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4861 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4921 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4981 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5041 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5101 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     5161 ggcgtatcac gaggcccttt cgtcttcac
//
