LOCUS       Exported                4381 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUCP24T, complete sequence.
ACCESSION   MF098686
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4381)
  AUTHORS   Lu Y, Zeng J, Wu B, E S, Wang L, Cai R, Zhang N, Li Y, Huang X, 
            Huang B, Chen C.
  TITLE     Quorum Sensing N-acyl Homoserine Lactones-SdiA Suppresses 
            Escherichia coli-Pseudomonas aeruginosa Conjugation through 
            Inhibiting traI Expression
  JOURNAL   Front Cell Infect Microbiol 7, 7 (2017)
  PUBMED    28164039
REFERENCE   2  (bases 1 to 4381)
  AUTHORS   Chen S, E S, Xiao Q, Zeng J, Lu Y, Ma X, He Y, Huang B, Chen C, 
            Zhang N, Cai R, Yuan H, Qu P, Lin D.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-MAY-2017) Department of Laboratory Medicine, The First
            Affiliated Hospital of Sun Yat-sen University, Zhongshan road, 
            Yuexiu District, Guangzhou, Guangdong 510000, China
REFERENCE   3  (bases 1 to 4381)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4381
                     /organism="Cloning vector pUCP24T"
                     /lab_host="Escherichia coli DH5 alpha"
                     /mol_type="other DNA"
                     /PCR_primers="fwd_name: orit-f, fwd_seq:
                     cccgggggaaggcagtacacctt, rev_name: orit-r, rev_seq: 
                     aagcttgggtagatattccacaaaacagcag"
                     /db_xref="taxon:1987384"
     rep_origin      46..397
                     /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)"
     gene            398..1244
                     /gene="rep"
                     /label=rep
     regulatory      398..401
                     /regulatory_class="ribosome_binding_site"
                     /gene="rep"
     CDS             411..1244
                     /codon_start=1
                     /gene="rep"
                     /product="replication protein"
                     /label=rep
                     /protein_id="ASN77034.1"
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     primer_bind     1405..1421
     oriT            1426..1811
                     /label=from RP4 plasmid
                     /note="from RP4 plasmid"
     oriT            1601..1710
                     /note="incP origin of transfer"
     primer_bind     1836..1852
     regulatory      1848..1851
                     /regulatory_class="ribosome_binding_site"
     protein_bind    1860..1876
                     /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)."
     regulatory      1884..1914
                     /regulatory_class="promoter"
     promoter        complement(1884..1914)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1929..1950
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2238..2826
     gene            complement(3627..4174)
                     /gene="aacC1"
                     /label=aacC1
     CDS             complement(3627..4160)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase-3-1"
                     /label=aacC1
                     /protein_id="ASN77035.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     regulatory      complement(4169..4174)
                     /regulatory_class="ribosome_binding_site"
                     /gene="aacC1"
     promoter        complement(4349..4377)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"