pUD1074 vector (V015815)

Basic Vector Information

Vector Name:
pUD1074
Antibiotic Resistance:
Ampicillin
Length:
7395 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Valk LC.

pUD1074 vector Map

pUD10747395 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300660069007200T7 promoterlac operatorRBS6xHisV5 tagTEV sitehybrid cluster proteinT7 terminatorAmpR promoterAmpRoribomropCAP binding sitelacIlacI promoter

pUD1074 vector Sequence

LOCUS       62056_22370        7395 bp DNA     circular SYN 28-JAN-2020
DEFINITION  Cloning vector pUD1074, complete sequence.
ACCESSION   MN498128
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7395)
  AUTHORS   Valk LC.
  TITLE     'Candidatus Galacturonibacter soehngenii' shows hetero-acetogenic 
            catabolism of galacturonic acid but lacks a canonical carbon 
            monoxide dehydrogenase/acetyl-CoA synthase complex
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7395)
  AUTHORS   Valk LC.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-SEP-2019) Biotechnology, Delft University of 
            Technology, Van der Maasweg 9, Delft, Zuid Holland 2629 HZ, 
            Netherlands
REFERENCE   3  (bases 1 to 7395)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-SEP-2019) Biotechnology, Delft University of Technology, Van der
            Maasweg 9, Delft, Zuid Holland 2629 HZ, Netherlands"
FEATURES             Location/Qualifiers
     source          1..7395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /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)."
     RBS             267..289
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             300..317
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             318..359
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
     CDS             360..380
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     CDS             396..2033
                     /codon_start=1
                     /transl_table=11
                     /product="hybrid cluster protein"
                     /label=hybrid cluster protein
                     /note="hydroxylamine reductase; putative novel carbon
                     monoxide dehydrogenase; codon optimized for Escherichia 
                     coli"
                     /protein_id="QHN70777.1"
                     /translation="MENKMFCYQCQETAGCTGCTQMGVCGKTPEVAGMQDLLVYVTKGL
                     AEVTTKLRKENVAIEKSTNHYITWNLFTTITNANFDEAVFVERINKSLDIKKELLAKLS
                     DKSGLSDAAIWEDYSVEGMRAKAAQVGVLATENEDVRSLRELITYGLKGLCAYMKHANA
                     LSKENEAVDIFVQETLAKLLDDSLSVNELIALTLETGKYGVDGMALLDEANTSSYGNPE
                     ITSVNIGVGKNPGILISGHDLRDMEMLLEQTKGTGVDVYTHSEMLPAHYYPAFKKYDHF
                     VGNYGNAWWKQKEEFESFNGPILMTTNCIVPPKASYKDRLFTTGSAGMEGCKHIEGNEG
                     EVKDFSEIIALAKTCQPPTEIETGSIVGGFAHNQVFALADKVVDAVKSGAIKKFVVMAG
                     CDGRAKSRNYYTDFAKALPNDSVILTAGCAKYKYNKLNLGDIGGIPRVLDAGQCNDSYS
                     LVLIALKLQEVFGLESVNDLPIAYNIAWYEQKAVIVLLSLLHLGVKNIHLGPTLPAFLS
                     PNVVNVLVENFGIAGITDVENDLKILLA"
     terminator      2110..2157
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2481..2585
                     /label=AmpR promoter
     CDS             2586..3443
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3617..4205
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4391..4530)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4635..4823)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(6101..6122)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(6138..7217)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(7218..7295)
                     /label=lacI promoter

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