LOCUS       40924_30640        4513 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMESy4, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4513)
  AUTHORS   Pardon E, Steyaert J.
  TITLE     A general protocol for the generation of Nanobodies for structural 
            biology
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4513)
  AUTHORS   Pardon E, Steyaert J.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-JUL-2013) Structural Biology Brussels, Vrije 
            Universiteit Brussel, Pleinlaan 2, Brussels 1050, Belgium
REFERENCE   3  (bases 1 to 4513)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4513)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-JUL-2013) Structural Biology Brussels, Vrije Universiteit 
            Brussel, Pleinlaan 2, Brussels 1050, Belgium"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     regulatory      211..215
                     /regulatory_class="ribosome_binding_site"
     sig_peptide     225..290
                     /label=pelB signal sequence
                     /note="leader peptide for secretion"
     CDS             340..357
                     /label=6xHis
                     /note="6xHis affinity tag"
     misc_feature    358..369
                     /label=CaptureSelect C-tag
                     /note="CaptureSelect C-tag"
     misc_feature    373..390
                     /label=HA-tag
                     /note="HA-tag"
     gene            403..1636
                     /gene="M13 geneIII"
                     /label=M13 geneIII
     primer_bind     complement(1642..1658)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1871..2326
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2608..2712
                     /label=AmpR promoter
     CDS             2713..3570
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3744..4332
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacaagctta aggagacagt acatatgaaa tacctattgc
      241 ctacggcagc cgctggattg ttattactcg cggcccagcc ggccatggcc caggtgcagc
      301 tgcaggagtc tagaggggac ccaggtcacc gtctcctcac accaccatca ccatcacgaa
      361 cctgaagcct agtacccgta cgacgttccg gactacggtt ccactgttga aagttgttta
      421 gcaaaacctc atacagaaaa ttcatttact aacgtctgga aagacgacaa aactttagat
      481 cgttacgcta actatgaggg ctgtctgtgg aatgctacag gcgttgtcgt ttgtactggt
      541 gacgaaactc agtgttacgg tacatgggtt cctattgggc ttgctatccc tgaaaatgag
      601 ggtggtggct ctgagggtgg cggttctgag ggtggcggtt ctgagggtgg cggtactaaa
      661 cctcctgagt acggtgatac acctattccg ggctatactt atatcaaccc tctcgacggc
      721 acttatccgc ctggtactga gcaaaacccc gctaatccta atccttctct tgaggagtct
      781 cagcctctta atactttcat gtttcagaat aataggttcc gaaataggca gggtgcatta
      841 actgtttata cgggcactgt tactcaaggc actgaccccg ttaaaactta ttaccagtac
      901 actcctgtat catcaaaagc catgtatgac gcttactgga acggtaaatt cagagactgc
      961 gctttccatt ctggctttaa tgaggatcca ttcgtttgtg aatatcaagg ccaatcgtct
     1021 gacctgcctc aacctcctgt caatgctggc ggcggctctg gtggtggttc tggtggcggc
     1081 tctgagggtg gcggctctga gggtggcggc tctgagggtg gcggttctga gggtggcggc
     1141 tctgagggtg gcggttccgg tggcggctcc ggttccggtg attttgatta tgaaaaaatg
     1201 gcaaacgcta ataagggggc tatgaccgaa aatgccgatg aaaacgcgct acagtctgac
     1261 gctaaaggca aacttgattc tgtcgctact gattacggtg ctgctatcga tggtttcatt
     1321 ggtgacgttt ccggccttgc taatggtaat ggtgctactg gtgattttgc tggctctaat
     1381 tcccaaatgg ctcaagtcgg tgacggtgat aattcacctt taatgaataa tttccgtcaa
     1441 tatttacctt ctttacctca gtcggttgaa tgtcgccctt atgtctttgg cgctggtaaa
     1501 ccatatgaat tttctattga ttgtgacaaa ataaacttat tccgtggtgt ctttgcgttt
     1561 cttttatatg ttgccacctt tatgtatgta ttttcgacgt ttgctaacat actgcgtaat
     1621 aaggagtctt aataagaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     1681 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     1741 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     1801 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     1861 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1921 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     1981 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2041 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     2101 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2161 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc gggctattct
     2221 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2281 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     2341 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     2401 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     2461 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     2521 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     2581 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2641 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2701 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     2761 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     2821 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2881 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2941 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3001 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3061 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3121 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3181 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3241 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3301 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3361 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3421 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3481 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3541 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3601 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3661 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3721 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3781 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3841 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     3901 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3961 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4021 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4081 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4141 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4201 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4261 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4321 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4381 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4441 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4501 gaggaagcgg aag
//
