pETcoco-2 vector (V010931)

LOCUS       pETcoco-2.       12417 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector with an ampicillin resistance marker that allows 
            single-copy replication, on-demand amplification, and tightly 
            regulated expression.
ACCESSION   .
VERSION     .
KEYWORDS    pETcoco-2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12417)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 12417)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     The copy number can be increased to 20-50 copies per cell by adding 
            arabinose to the growth medium.
FEATURES             Location/Qualifiers
     source          1..12417
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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             42..64
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             72..74
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             75..92
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             102..146
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             162..176
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             249..281
                     /label=HSV tag
                     /note="HSV (herpes simplex virus) epitope tag"
     CDS             326..343
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      410..457
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(834..1709)
                     /label=araC
                     /note="L-arabinose regulatory protein"
     promoter        1736..2020
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     CDS             2080..3225
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     CDS             complement(3449..4306)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4307..4398)
                     /label=AmpR promoter
     rep_origin      complement(4965..5677)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      5756..5975
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             6066..6818
                     /label=repE
                     /note="replication initiation protein for the bacterial F 
                     plasmid"
     misc_feature    6824..7074
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             7400..8572
                     /label=sopA
                     /note="partitioning protein for the bacterial F plasmid"
     CDS             8575..9543
                     /label=sopB
                     /note="partitioning protein for the bacterial F plasmid"
     misc_feature    9619..10092
                     /gene="sopC"
                     /label=sopC
                     /note="sopC"
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     misc_feature    complement(10352..10750)
                     /label=cos
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
     protein_bind    complement(10768..10801)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(10894..10915)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(10931..12010)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(12011..12088)
                     /label=lacI promoter