LOCUS       Exported                5087 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Escherichia-Pseudomonas shuttle vector pHERD20T, complete sequence.
ACCESSION   EU603324
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5087)
  AUTHORS   Qiu D, Damron FH, Mima T, Schweizer HP, Yu HD.
  TITLE     PBAD-based shuttle vectors for functional analysis of toxic and 
            highly regulated genes in Pseudomonas and Burkholderia spp. and 
            other bacteria
  JOURNAL   Appl. Environ. Microbiol. 74 (23), 7422-7426 (2008)
  PUBMED    18849445
REFERENCE   2  (bases 1 to 5087)
  AUTHORS   Qiu D, Yu HD.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-MAR-2008) Biochemistry and Microbiology, Marshall 
            University Joan C. Edwards School of Medicine, 1 John Marshall 
            Drive, Huntington, WV 25755, USA
REFERENCE   3  (bases 1 to 5087)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5087
                     /organism="Escherichia-Pseudomonas shuttle vector
                     pHERD20T"
                     /mol_type="other DNA"
                     /note="conjugation-based shuttle vector with carbenicillin 
                     resistance marker"
                     /db_xref="taxon:544738"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             201..1061
                     /codon_start=1
                     /gene="bla"
                     /product="Bla"
                     /label=bla
                     /note="beta-lactamase"
                     /protein_id="ACF17853.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            201..1061
                     /gene="bla"
                     /label=bla
     rep_origin      1216..1835
                     /label=ori_pBR322
                     /note="ori_pBR322"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     oriT            1885..2152
     oriT            1892..2000
                     /note="incP origin of transfer"
     CDS             complement(2157..3035)
                     /codon_start=1
                     /gene="araC"
                     /product="AraC"
                     /label=araC
                     /note="arabinose operon regulator"
                     /protein_id="ACF17856.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            complement(2157..3035)
                     /gene="araC"
                     /label=araC
     promoter        3062..3346
                     /gene="araBAD"
                     /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)"
     RBS             3373..3395
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3407..3661
                     /codon_start=1
                     /gene="lacZalpha"
                     /product="LacZalpha"
                     /label=lacZalpha
                     /note="beta galactosidase alpha peptide"
                     /protein_id="ACF17854.1"
                     /translation="MGSDKNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLKGRPGRGGHGL"
     gene            3407..3661
                     /gene="lacZalpha"
                     /label=lacZalpha
     misc_feature    3421..3477
                     /label=multiple cloning site
                     /note="multiple cloning site"
     primer_bind     complement(3481..3497)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3658..4491)
                     /codon_start=1
                     /gene="rep"
                     /product="Rep"
                     /label=rep
                     /note="replication protein"
                     /protein_id="ACF17855.1"
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     gene            complement(3658..4491)
                     /gene="rep"
                     /label=rep
     rep_origin      4492..4901
                     /label=ori_pRO1600
                     /note="ori_pRO1600"
     rep_origin      4505..4856
                     /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)"