LOCUS       40924_38963        4575 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pSCCA5-E8d DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4575)
  AUTHORS   Arimitsu H, Sasaki K, Iba Y, Kurosawa Y, Shimizu T, Tsuji T.
  TITLE     Isolation of B subunit-specific monoclonal antibody clones that 
            strongly neutralize the toxicity of Shiga toxin 2
  JOURNAL   Microbiol. Immunol. 59 (2), 71-81 (2015)
  PUBMED    25521016
REFERENCE   2  (bases 1 to 4575)
  AUTHORS   Arimitsu H, Iba Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JUN-2014) Contact:Hideyuki Arimitsu Fujita Health 
            University, Department of Microbiology; 1-98, Dengakugakubo, 
            Kutsukake-cho, Toyoake, Aichi 470-1192, Japan URL 
            :https://fujita-hu.ac.jp/
REFERENCE   3  (bases 1 to 4575)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4575)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microbiol. 
            Immunol."; date: "2015"; volume: "59"; issue: "2"; pages: "71-81"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-JUN-2014) Contact:Hideyuki Arimitsu Fujita Health University, 
            Department of Microbiology"; volume: " 1-98, Dengakugakubo, 
            Kutsukake-cho, Toyoake, Aichi 470-1192, Japan URL :https"; pages: 
            "//fujita-hu.ac.jp"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    40..87
                     /label=PelB leader sequence
                     /note="PelB leader sequence"
     misc_feature    197..241
                     /label=Single chain linker
                     /note="Single chain linker"
     CDS             375..692
                     /codon_start=1
                     /label=mIg-kappa-CL
                     /note="Mouse immunoglobulin kappa light chain constant
                     region"
                     /translation="ADAAPTVSIFPPSSEQLTSGGASVVCFLNSFYPKDINVKWKIDGS
                     ERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRN
                     EC"
     misc_feature    711..1340
                     /label=cp3
                     /note="cp3"
     CDS             1393..1566
                     /codon_start=1
                     /label=ProtA
                     /note="IgG-binding unit of Staphylococcus aureus protein A"
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             1570..1740
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     primer_bind     complement(1770..1786)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(1928..2383)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2410..2514
                     /label=AmpR promoter
     CDS             2515..3372
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3546..4134
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4422..4443
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4458..4488
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4496..4512
                     /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)."
     primer_bind     4520..4536
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4555..4573
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aagcttgcat gcaaattcta tttcaaggag acagtcataa tgaaatacct attgcctacg
       61 gcagccgctg gattgttatt actcgcggcc cagccggccc ctgacatctg aggacactgc
      121 cgtctattac tgtgctggtt atgattacgg caactttgac tactggggcc aaggcaccac
      181 ggtcaccgtc tcgagcggcc gcggtggttc tggtggtggc ggttctggcg gtggtggttc
      241 catggccgac atcgagctca cccagtctcc agcctccctt tctgcgtctg tgggagaaac
      301 tgtcaccatc acatgtcgag caagtgggaa tattcacaat tatttagcat ggtaccaagc
      361 tcgagatcaa acgggctgat gctgcaccaa ctgtatccat cttcccacca tccagtgagc
      421 agttaacatc tggaggtgcc tcagtcgtgt gcttcttgaa cagcttctac cccaaagaca
      481 tcaatgtcaa gtggaagatt gatggcagtg aacgacaaaa tggcgtcctg aacagttgga
      541 ctgatcagga cagcaaagac agcacctaca gcatgagcag caccctcacg ttgaccaagg
      601 acgagtatga acgacataac agctatacct gtgaggccac tcacaagaca tcaacttcac
      661 ccattgtcaa gagcttcaac aggaatgagt gttcggcgcg ccagtcgact ccattcgttt
      721 gtgaatatca aggccaatcg tctgacctgc ctcaacctcc tgtcaatgct ggcggcggct
      781 ctggtggtgg ttctggtggc ggctctgagg gtggtggctc tgagggtggc ggttctgagg
      841 gtggcggctc tgagggaggc ggttccggtg gtggctctgg ttccggtgat tttgattatg
      901 aaaagatggc aaacgctaat aagggggcta tgaccgaaaa tgccgatgaa aacgcgctac
      961 agtcagacgc taaaggcaaa cttgattctg tcgctactga ttacggtgct gctatcgatg
     1021 gtttcattgg tgacgtttcc ggccttgcta atggtaatgg tgctactggt gattttgctg
     1081 gctctaattc ccaaatggct caagtcggtg acggtgataa ttcaccttta atgaataatt
     1141 tccgtcaata tttaccttcc ctccctcaat cggttgaatg tcgccctttt gtctttggcg
     1201 ctggtaaacc atatgaattt tctattgatt gtgacaaaat aaacttattc cgtggtgtct
     1261 ttgcgtttct tttatatgtt gccaccttta tgtatgtatt ttctacgttt gctaacatac
     1321 tgcgtaataa ggagtcttaa tcatgccagt tcttttgggt gctagctgtc gactgcgcaa
     1381 cacgatgaag ccgtagacaa caaattcaac aaagaacaac aaaacgcgtt ctatgagatc
     1441 ttacatttac ctaacttaaa cgaagaacaa cgaaacgcct tcatccaaag tttaaaagat
     1501 gacccaagcc aaagcgctaa ccttttagca gaagctaaaa agctaaatga tgctcaggcg
     1561 ccgaaagtag acaacaaatt caacaaagaa caacaaaacg cgttctatga gatcttacat
     1621 ttacctaact taaacgaaga acaacgaaac gccttcatcc aaagtttaaa agatgaccca
     1681 agccaaagcg ctaacctttt agcagaagct aaaaagctaa atgatgctca ggcgccgaaa
     1741 gtagacgcga attagctggg aattaattca ctggccgtcg ttttacaacg tcgtgactgg
     1801 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     1861 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     1921 gaatgggaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     1981 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     2041 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     2101 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     2161 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     2221 gggatgcccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     2281 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     2341 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     2401 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2461 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2521 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2581 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2641 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     2701 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     2761 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2821 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2881 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     2941 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3001 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3061 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3121 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3181 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3241 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3301 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3361 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     3421 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3481 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3541 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3601 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3661 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3721 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3781 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3841 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3901 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc
     3961 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4021 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4081 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4141 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     4201 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     4261 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     4321 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     4381 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     4441 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     4501 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctctaatac
     4561 gactcactat aggga
//
