LOCUS       62056_16750        4434 bp DNA     circular SYN 07-JAN-2021
DEFINITION  Cloning vector pMESD22c7, complete sequence.
ACCESSION   MT338520
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4434)
  AUTHORS   Uchanski T, Masiulis S, Fischer B, Kalichuk V, Lopez-Sanchez U, 
            Zarkadas E, Weckener M, Sente A, Ward P, Wohlkoenig A, Zoegg T, 
            Remaut H, Naismith JH, Nury H, Vranken W, Aricescu AR, Pardon E, 
            Steyaert J.
  TITLE     Megabodies expand the nanobody toolkit for protein structure 
            determination by single-particle cryo-EM
  JOURNAL   Nat Methods 18 (1), 60-68 (2021)
REFERENCE   2  (bases 1 to 4434)
  AUTHORS   Steyaert J, Uchanski T, Pardon E.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-APR-2020) VIB-VUB Center for Structural Biology, VIB, 
            Pleinlaan 2, Brussels 1050, Belgium
REFERENCE   3  (bases 1 to 4434)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods"; date: "2021"; volume: "18"; issue: "1"; pages: "60-68"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (14-APR-2020) VIB-VUB Center for Structural Biology, VIB, Pleinlaan 
            2, Brussels 1050, Belgium"
FEATURES             Location/Qualifiers
     source          1..4434
                     /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)."
     misc_feature    225..281
                     /label=DsbA signal peptide
                     /note="DsbA signal peptide"
     misc_feature    282..320
                     /label=nanobody beta-strain A (residues 1-14)
                     /note="nanobody beta-strain A (residues 1-14)"
     misc_feature    321..1487
                     /label=c7HopQ scaffold protein
                     /note="c7HopQ scaffold protein"
     misc_feature    1488..1490
                     /label=nanobody residue 17
                     /note="nanobody residue 17"
     misc_feature    1491..1523
                     /label=multi cloning site
                     /note="multi cloning site"
     CDS             1524..1541
                     /label=6xHis
                     /note="6xHis affinity tag"
     misc_feature    1542..1553
                     /label=EPEA-tag
                     /note="EPEA-tag"
     primer_bind     complement(1563..1579)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1792..2247
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2529..2633
                     /label=AmpR promoter
     CDS             2634..3491
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3665..4253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agagcccaat 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 aagatttggc
      241 tggcgctggc tggtttagtt ttagcgttta gcgcatcggc gcaggtgcag ctggtggagt
      301 ctgggggagg actagtacag actaaaacaa caacttctgt tattgatact actaatgatg
      361 cccaaaatct tttgactcaa gcgcaaacta ttgtcaatac ccttaaagat tattgcccta
      421 tcttgatcgc aaaatctagt agtagcaatg gtggcacaaa caatgctaac accccttcat
      481 ggcaaacagc cggtggcggc aaaaattcat gcgctacttt tggtgcagag tttagtgccg
      541 cttcagacat gattaataat gcacaaaaaa tcgttcaaga aacccaacaa ctgagcgcca
      601 accaaccaaa aaatatcaca caacctcata atctgaacct taacagccct agcagtctta
      661 ccgctttagc tcaaaaaatg ctgaaaaacg cgcaatctca agcagaaatt ttaaaacttg
      721 ccaatcaagt ggagagcgat tttaacaaac tttcttcagg ccatcttaaa gactacattg
      781 ggaaatgtga cgcctctgct atcagtagtg ctaatatgac aatgcagaat cagaagaaca
      841 actgggggaa cgggtgcgct ggcgtggaag aaactcagtc actgttaaaa acaagcgccg
      901 ctgattttaa caaccaaact cctcaaatca atcaagctca aaacctggcc aacaccctta
      961 ttcaagaact tggcaacaac acttatgaac agctaagccg tcttttaacc aacgataatg
     1021 gcacaaactc aaagacaagc gcccaagcta tcaaccaagc ggttaataat ttgaacgaac
     1081 gtgcaaaaac tttagccggt gggacaacca attctccggc atatcaagcc acgcttttag
     1141 ccttacgttc tgtgttaggg ctgtggaata gcatgggtta tgctgtcatc tgcggaggtt
     1201 ataccaaaag tccaggcgaa aacaatcaaa aagatttcca ctacaccgat gagaatggca
     1261 acggcactac aatcaattgc ggtgggagca caaatagtaa tggcactcat agttataatg
     1321 gcacaaatac attaaaagca gacaaaaatg tttctctttc tattgagcaa tatgaaaaaa
     1381 tccatgaagc ctatcagatt ctttcaaaag ccttaaaaca agccgggctt gctcctttaa
     1441 atagcaaagg ggaaaagtta gaagcacatg taaccacatc aaagtatgga tccggaagag
     1501 cttagtcgac aagctcttca agtcaccacc atcaccatca cgaacctgaa gcctaagaat
     1561 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     1621 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     1681 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     1741 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     1801 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     1861 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     1921 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     1981 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     2041 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2101 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
     2161 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2221 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     2281 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     2341 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     2401 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     2461 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     2521 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2581 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     2641 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2701 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     2761 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2821 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     2881 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2941 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3001 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3061 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3121 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3181 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3241 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3301 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3361 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3421 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3481 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     3541 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3601 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3661 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3721 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3781 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     3841 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3901 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3961 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4021 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4081 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4141 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4201 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4261 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4321 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4381 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaag
//
