Basic Vector Information
- Vector Name:
- pACYC_synB_cstII_lgtB_ApNGT
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 8180 bp
- Type:
- Expression vector
- Replication origin:
- p15A ori
- Source/Author:
- 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.
pACYC_synB_cstII_lgtB_ApNGT vector Vector Map
pACYC_synB_cstII_lgtB_ApNGT vector Sequence
LOCUS V009585 8180 bp DNA circular SYN 17-DEC-2018
DEFINITION Exported.
ACCESSION V009585
VERSION V009585
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8180)
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 8180)
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 8180)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8180)
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..8180
/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..4342
/codon_start=1
/product="SynB"
/label="SynB"
/protein_id="ATN32179.1"
/translation="MEKQNIAVILARQNSKGLPLKNLRKMNGISLLGHTINAAISSKCF
DRIIVSTDGGLIAEEAKNFGVEVVLRPAELASDTASSISGVIHALETIGSNSGTVTLLQ
PTSPLRTGAHIREAFSLFDEKIKGSVVSACPMEHHPLKTLLQINNGEYAPMRHLSDLEQ
PRQQLPQAFRPNGAIYINDTASLIANNCFFIAPTKLYIMSHQDSIDIDTELDLQQAENI
LNHKES"
promoter 4357..4387
/label="lac UV5 promoter"
/note="E. coli lac promoter with an ""up"" mutation"
regulatory 4357..4362
/regulatory_class="minus_35_signal"
regulatory 4381..4387
/note="-10 seq (TATAAT; Pribnow box)"
/regulatory_class="minus_10_signal"
protein_bind 4391..4415
/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 4393..4413
/label="lac operator"
/note="lac operator"
CDS 4463..5242
/codon_start=1
/product="CstII"
/label="CstII"
/note="delta-C32 and I53S mutation"
/protein_id="ATN32180.1"
/translation="MKKVIIAGNGPSLKEIDYSRLPNDFDVFRCNQFYFEDKYYLGKKC
KAVFYNPSLFFEQYYTLKHLIQNQEYETELIMCSNYNQAHLENENFVKTFYDYFPDAHL
GYDFFKQLKDFNAYFKFHEIYFNQRITSGVYMCAVAIALGYKEIYLSGIDFYQNGSSYA
FDTKQKNLLKLAPNFKNDNSHYIGHSKNTDIKALEFLEKTYKIKLYCLCPNSLLANFIG
LAPNLNSNFIIQEKNNYTKDILIPSSEAYGKFSKNIN"
promoter 5254..5284
/label="lac UV5 promoter"
/note="E. coli lac promoter with an ""up"" mutation"
regulatory 5254..5259
/regulatory_class="minus_35_signal"
regulatory 5278..5284
/note="-10 seq (TATAAT; Pribnow box)"
/regulatory_class="minus_10_signal"
protein_bind 5288..5312
/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 5290..5310
/label="lac operator"
/note="lac operator"
CDS 5360..6190
/codon_start=1
/product="LgtB"
/label="LgtB"
/protein_id="ATN32181.1"
/translation="MSQNHVISLASAAERRAHIADTFGRHGIPFQFFDALMPSERLEQA
MAELVPGLSAHPYLSGVEKACFMSHAVLWKQALDEGLPYITVFEDDVLLGEGAEKFLAE
DAWLQERFDPDTAFIVRLETMFMHVLTSPSGVADYCGRAFPLLESEHWGTAGYIISRKA
MRFFLDRFAALPPEGLHPVDLMMFSDFFDREGMPVCQLNPALCAQELHYAKFHDQNSAL
GSLIEHDRLLNRKQQRRDSPANTFKHRLIRALTKISREREKRRQRREQFIVPFQ"
promoter 6202..6232
/label="lac UV5 promoter"
/note="E. coli lac promoter with an ""up"" mutation"
regulatory 6202..6207
/regulatory_class="minus_35_signal"
regulatory 6226..6232
/note="-10 seq (TATAAT; Pribnow box)"
/regulatory_class="minus_10_signal"
protein_bind 6236..6260
/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 6238..6258
/label="lac operator"
/note="lac operator"
CDS 6308..8167
/note="UDP-glucose:protein N-beta-glucosyltransferase from
Actinobacillus pleuropneumoniae serotype 7 (strain AP76).
Accession#: B3H2N2"
/label="UDP-glucose:protein N-beta-glucosyltransferase"
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