LOCUS       40924_28631        4500 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Suicide vector pLP11, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4500)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2015"; volume: "10"; issue: "12"; pages: "E0144465"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "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"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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      complement(125..513)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(641..1297)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /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"
     oriT            1699..1808
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        2079..2156
                     /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..3236
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    3252..3273
                     /label=CAP binding site
                     /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)"
     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
                     /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"
ORIGIN
        1 ctcgagggaa tataagtcga cggatccccc gggctgcagg aatctagacc ttgagtcgaa
       61 ttcgagctcg acatatcgtc gctagctgtt ccctttagtg agggttaatt gcgcgcaatt
      121 cccatgtcag ccgttaagtg ttcctgtgtc actcaaaatt gctttgagag gctctaaggg
      181 cttctcagtg cgttacatcc ctggcttgtt gtccacaacc gttaaacctt aaaagcttta
      241 aaagccttat atattctttt ttttcttata aaacttaaaa ccttagaggc tatttaagtt
      301 gctgatttat attaatttta ttgttcaaac atgagagctt agtacgtgaa acatgagagc
      361 ttagtacgtt agccatgaga gcttagtacg ttagccatga gggtttagtt cgttaaacat
      421 gagagcttag tacgttaaac atgagagctt agtacgtgaa acatgagagc ttagtacgta
      481 ctatcaacag gttgaactgc tgatcttcag atcctctacg ccggacgcat cgtggccgga
      541 tcagatctga tatcgtcgca gaccaaaacg atctcaagaa gatcatctta ttaatcagat
      601 aaaatatttc taggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat
      661 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
      721 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
      781 tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga
      841 aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat
      901 aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga
      961 aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg
     1021 tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa
     1081 attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt
     1141 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat
     1201 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata
     1261 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa
     1321 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     1381 aacctcttac gtgccgatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg
     1441 tatcaacagg gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtattt
     1501 attcggcgca attgatatcg cgcgcgtaat acgactcact atagggcgaa ttgggtacca
     1561 gcgcttttcc gctgcataac cctgcttcgg ggtcattata gcgatttttt cggtatatcc
     1621 atcctttttc gcacgatata caggattttg ccaaagggtt cgtgtagact ttccttggtg
     1681 tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc
     1741 cttcttcact gtcccttatt cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg
     1801 ctggccggcg tcgacggtat cgataagctt ggcatgcctg cagcatacac tccgctatcg
     1861 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     1921 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     1981 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagccgga tcaattctca
     2041 tgtttgacag cttatcatcg agcttactcc ccatccccga caccatcgaa tggtgcaaaa
     2101 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga
     2161 aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc
     2221 gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga
     2281 tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt
     2341 tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg
     2401 cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa
     2461 gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc
     2521 tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta
     2581 atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt attattttct
     2641 cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa
     2701 tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc
     2761 ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg
     2821 ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga
     2881 tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc
     2941 tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt
     3001 atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg
     3061 accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct
     3121 cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt
     3181 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     3241 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     3301 cttccgggtt ttctttgcct cacgatcgcc cccaaaacac ataaccaatt gtatttattg
     3361 aaaaataaat agatacaact cactaaacat agcaattcag atccggtcca gtaatgacct
     3421 cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg
     3481 agaatcgcag caacttgtcg cgccaatcga gccatgtcgt cgtcaacgac cccccattca
     3541 agaacagcaa gcagcattga gaactttgga atccagtccc tcttccacct gctgatccgg
     3601 atcgatcccg ttatcgactg cacggtgcac caatgcttct ggcgtcaggc agccatcgga
     3661 agctgtggta tggctgtgca ggtcgtaaat cactgcataa ttcgtgtcgc tcaaggcgca
     3721 ctcccgttct ggataatgtt ttttgcgccg acatcataac ggttctggca aatattctga
     3781 aatgagctgt tgacaattaa tcatcggctc gtataatgtg tggaattgtg agcggataac
     3841 aatttcacac aggaaacaga attgatccgg ccaagcttgt aagaaggaga tatacatatg
     3901 atgaccaaaa aacctgaatt ttatgctcca gacctaacgg aagaacgctt gcacatattg
     3961 agtgaaaacc tactcgatgt gctcgatgag gcccatcatt actcagaaag tcccaatgcg
     4021 acggcctggt ttaaaggcac tgcaaattat ggtttaccac agggaatgct aatccgtatg
     4081 cactcagaca gtgcttatcc atggctcaca cttgccaacc aaacaatgga ttacactgct
     4141 agagttggaa acactcttgt tcaatttgtt gtggatgacc ctcactcacc tcgcaagcaa
     4201 catcgcctta aacgcaatgc tgttgaaaag catcaaatat ctcttgaatt agaggaagac
     4261 tatgttgata ccccattgat ttggcgattt tacttaaatc ctatatctaa tggtgttgat
     4321 tattccccat ctatttcttt gttgggtttc aacggcaatg gcaacgttat atgcagttgg
     4381 gaatacgaca cagttgtaac tggtccagtt gttacagaca aaccagaatc ggttgaaatt
     4441 gatgagccat tattggttcg taaaaagaaa gtacaaaaaa aggtctctga tgagtaaaag
//
