pACYC_lgtB_ApNGT vector (V009586)

LOCUS       V009586                 6476 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009586
VERSION     V009586
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6476)
  AUTHORS   Keys TG, Wetter M, Hang I, Rutschmann C, Russo S, Mally M, Steffen
            M, Zuppiger M, Muller F, Schneider J, Faridmoayer A, Lin CW, Aebi M.
  TITLE     A biosynthetic route for polysialylating proteins in Escherichia
            coli
  JOURNAL   Metab. Eng. (2017) In press
   PUBMED   29101090
REFERENCE   2  (bases 1 to 6476)
  AUTHORS   Keys TG.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-NOV-2016) Institute of Microbiology, ETH Zurich,
            Vladimir-Prelog-Weg 4, Zurich, ZH 8093, Switzerland
REFERENCE   3  (bases 1 to 6476)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6476)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Metab. Eng.
            (2017) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (21-NOV-2016) Institute of Microbiology, ETH Zurich,
            Vladimir-Prelog-Weg 4, Zurich, ZH 8093, Switzerland"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      28..75
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             complement(298..954)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(955..1057)
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(1583..2127)
                     /direction=LEFT
                     /label="p15A ori"
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells
                     that contain a second plasmid with the ColE1 origin."
     protein_bind    complement(2318..2339)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2355..3434)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(3435..3512)
                     /label="lacI promoter"
     promoter        3550..3580
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    3584..3608
                     /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)."
     CDS             3656..4486
                     /codon_start=1
                     /product="LgtB"
                     /label="LgtB"
                     /protein_id="ATN32183.1"
                     /translation="MSQNHVISLASAAERRAHIADTFGRHGIPFQFFDALMPSERLEQA
                     MAELVPGLSAHPYLSGVEKACFMSHAVLWKQALDEGLPYITVFEDDVLLGEGAEKFLAE
                     DAWLQERFDPDTAFIVRLETMFMHVLTSPSGVADYCGRAFPLLESEHWGTAGYIISRKA
                     MRFFLDRFAALPPEGLHPVDLMMFSDFFDREGMPVCQLNPALCAQELHYAKFHDQNSAL
                     GSLIEHDRLLNRKQQRRDSPANTFKHRLIRALTKISREREKRRQRREQFIVPFQ"
     promoter        4498..4528
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an ""up"" mutation"
     regulatory      4498..4503
                     /regulatory_class="minus_35_signal"
     regulatory      4522..4528
                     /note="-10 seq (TATAAT; Pribnow box)"
                     /regulatory_class="minus_10_signal"
     protein_bind    4532..4556
                     /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)."
     misc_feature    4534..4554
                     /label="lac operator"
                     /note="lac operator"
     CDS             4604..6463
                     /note="UDP-glucose:protein N-beta-glucosyltransferase from
                     Actinobacillus pleuropneumoniae serotype 7 (strain AP76).
                     Accession#: B3H2N2"
                     /label="UDP-glucose:protein N-beta-glucosyltransferase"