LOCUS       Exported                5045 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUCP20T-eYFP, complete sequence.
ACCESSION   KT878730
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5045)
  AUTHORS   Barbier M, Damron FH.
  TITLE     Rainbow Vectors for Broad-Range Bacterial Fluorescence Labeling
  JOURNAL   PLoS ONE 11 (3), E0146827 (2016)
  PUBMED    26937640
REFERENCE   2  (bases 1 to 5045)
  AUTHORS   Barbier M, Damron FH.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-OCT-2015) Microbiology, Immunology and Cell Biology, 
            West Virginia University, School of Medicine, One Medical Center 
            Drive, Morgantown, WV 26506, USA
REFERENCE   3  (bases 1 to 5045)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5045
                     /organism="Cloning vector pUCP20T-eYFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1811351"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             201..1061
                     /codon_start=1
                     /gene="bla"
                     /product="Bla"
                     /label=bla
                     /protein_id="AMO27226.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            201..1061
                     /gene="bla"
                     /label=bla
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     oriT            1892..2000
                     /note="incP origin of transfer"
     protein_bind    2384..2405
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2420..2450
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2458..2474
                     /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     2482..2498
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             2538..2549
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2556..3275
                     /codon_start=1
                     /gene="eyfp"
                     /product="eYFP"
                     /label=eyfp
                     /protein_id="AMO27227.1"
                     /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     gene            2556..3275
                     /gene="eyfp"
                     /label=eyfp
     terminator      3298..3392
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(3439..3455)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3616..4449)
                     /codon_start=1
                     /gene="rep"
                     /product="Rep"
                     /label=rep
                     /protein_id="AMO27225.1"
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     gene            complement(3616..4449)
                     /gene="rep"
                     /label=rep
     rep_origin      4463..4814
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from 
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"