LOCUS       Exported                4500 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Suicide vector pLP11, complete sequence.
ACCESSION   KT326152
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4500)
  AUTHORS   Luo P, He X, Liu Q, Hu C.
  TITLE     Developing Universal Genetic Tools for Rapid and Efficient Deletion 
            Mutation in Vibrio Species Based on Suicide T-Vectors Carrying a 
            Novel Counterselectable Marker, vmi480
  JOURNAL   PLoS ONE 10 (12), E0144465 (2015)
  PUBMED    26641275
REFERENCE   2  (bases 1 to 4500)
  AUTHORS   Luo P, He X, Hu C, Liu Q.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-JUL-2015) Key Laboratory of Marine Bio-resources 
            Sustainable Utilization, South China Sea Institute of Oceanology, 
            Xingang Xi Road 164, Guangzhou, Guangdong 510301, China
REFERENCE   3  (bases 1 to 4500)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4500
                     /organism="Suicide vector pLP11"
                     /mol_type="other DNA"
                     /db_xref="taxon:1769048"
     misc_feature    47..86
                     /label=multiple cloning sites
                     /note="multiple cloning sites"
     promoter        complement(92..110)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     rep_origin      125..513
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     rep_origin      138..564
                     /label=origin of replication
                     /note="origin of replication"
     CDS             complement(638..1297)
                     /codon_start=1
                     /product="chloramphenicol-resistant protein"
                     /label=chloramphenicol-resistant protein
                     /note="cat"
                     /protein_id="ALS55899.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(1298..1400)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     promoter        1527..1545
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(1559..1810)
                     /label=RP4 transfer origin
                     /note="RP4 transfer origin"
     oriT            1699..1808
                     /note="incP origin of transfer"
     promoter        2079..2156
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             2157..3239
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     CDS             2280..3239
                     /codon_start=1
                     /product="repressor protein"
                     /label=repressor protein
                     /note="lacIq"
                     /protein_id="ALS55900.1"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSR
                     ADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPA
                     LFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKY
                     LTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLR
                     VGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVS
                     LVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ"
     protein_bind    3252..3273
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      3441..3475
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     regulatory      3781..3824
                     /regulatory_class="promoter"
     promoter        3790..3818
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3826..3842
                     /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)."
     CDS             3901..4497
                     /codon_start=1
                     /product="toxin vmi480"
                     /label=toxin vmi480
                     /protein_id="ALS55901.1"
                     /translation="MTKKPEFYAPDLTEERLHILSENLLDVLDEAHHYSESPNATAWFK
                     GTANYGLPQGMLIRMHSDSAYPWLTLANQTMDYTARVGNTLVQFVVDDPHSPRKQHRLK
                     RNAVEKHQISLELEEDYVDTPLIWRFYLNPISNGVDYSPSISLLGFNGNGNVICSWEYD
                     TVVTGPVVTDKPESVEIDEPLLVRKKKVQKKVSDE"