LOCUS       Exported                3655 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pR18GFP, complete sequence.
ACCESSION   KP701018
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3655)
  AUTHORS   Cheng M, Zhang H, Guo S, He J, Li S, Huang X.
  TITLE     Screening Transcription Factor Genes Involved in Bacterial 
            Xenobiotic Catabolism using Modified SIGEX Approach
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3655)
  AUTHORS   Cheng M, Zhang H, Guo S, He J, Li S, Huang X.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-JAN-2015) Department of Microbiology, College of Life 
            Sciences, Nanjing Agricultural University, Tongwei Road #6, Nanjing,
            Jiangsu 210095, China
REFERENCE   3  (bases 1 to 3655)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3655
                     /organism="Cloning vector pR18GFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1663166"
     regulatory      47..94
                     /regulatory_class="terminator"
     terminator      47..94
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      306..311
                     /regulatory_class="ribosome_binding_site"
     CDS             319..1038
                     /codon_start=1
                     /product="eGFP"
                     /label=eGFP
                     /note="green fluorescent protein"
                     /protein_id="AKL72305.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(1185..1508)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     1418..1434
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1438..1494
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1504..1520)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1528..1544
                     /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)."
     promoter        complement(1552..1582)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1597..1618
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1906..2494)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2665..3525)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3526..3630)
                     /gene="bla"
                     /label=AmpR promoter