LOCUS       Exported                6465 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLac-GFP, complete sequence.
ACCESSION   KP294375
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6465)
  AUTHORS   Swofford CA, Van Dessel N, Forbes NS.
  TITLE     Quorum-sensing Salmonella selectively trigger protein expression 
            within tumors
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 112 (11), 3457-3462 (2015)
  PUBMED    25737556
REFERENCE   2  (bases 1 to 6465)
  AUTHORS   Swofford CA, Van Dessel N, Forbes NS.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-DEC-2014) Chemical Engineering, University of 
            Massachusetts Amherst, 686 North Pleasant Street, Amherst, MA 
            01003-9303, USA
REFERENCE   3  (bases 1 to 6465)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6465
                     /organism="Cloning vector pLac-GFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1628426"
     protein_bind    133..154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        169..199
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     regulatory      169..198
                     /regulatory_class="promoter"
                     /note="lac"
     protein_bind    207..223
                     /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)."
     primer_bind     231..247
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             275..991
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=green fluorescent protein
                     /note="GFPmut3"
                     /protein_id="AJT39559.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     oriT            1615..1702
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      3168..3850
                     /label=ColE1
                     /note="ColE1"
     rep_origin      complement(3186..3774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3945..4805)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(3945..4607)
                     /codon_start=1
                     /product="ampicillin resistance protein"
                     /label=ampicillin resistance protein
                     /protein_id="AJT39558.1"
                     /translation="MSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     promoter        complement(4806..4910)
                     /gene="bla"
                     /label=AmpR promoter