LOCUS       40924_28636        3871 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Suicide vector pLP12, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3871)
  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 3871)
  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 3871)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3871)
  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..3871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             21..617
                     /codon_start=1
                     /product="toxin vmi480"
                     /label=toxin vmi480
                     /protein_id="ALS55902.1"
                     /translation="MTKKPEFYAPDLTEERLHILSENLLDVLDEAHHYSESPNATAWFK
                     GTANYGLPQGMLIRMHSDSAYPWLTLANQTMDYTARVGNTLVQFVVDDPHSPRKQHRLK
                     RNAVEKHQISLELEEDYVDTPLIWRFYLNPISNGVDYSPSISLLGFNGNGNVICSWEYD
                     TVVTGPVVTDKPESVEIDEPLLVRKKKVQKKVSDE"
     misc_feature    667..706
                     /label=multiple cloning sites
                     /note="multiple cloning sites"
     promoter        complement(712..730)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     rep_origin      complement(745..1133)
                     /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(1261..1917)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(1918..2020)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     promoter        2147..2165
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     oriT            2319..2428
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(2671..3546)
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3573..3857
                     /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)"
ORIGIN
        1 aagaaggaga tatacatatg atgaccaaaa aacctgaatt ttatgctcca gacctaacgg
       61 aagaacgctt gcacatattg agtgaaaacc tactcgatgt gctcgatgag gcccatcatt
      121 actcagaaag tcccaatgcg acggcctggt ttaaaggcac tgcaaattat ggtttaccac
      181 agggaatgct aatccgtatg cactcagaca gtgcttatcc atggctcaca cttgccaacc
      241 aaacaatgga ttacactgct agagttggaa acactcttgt tcaatttgtt gtggatgacc
      301 ctcactcacc tcgcaagcaa catcgcctta aacgcaatgc tgttgaaaag catcaaatat
      361 ctcttgaatt agaggaagac tatgttgata ccccattgat ttggcgattt tacttaaatc
      421 ctatatctaa tggtgttgat tattccccat ctatttcttt gttgggtttc aacggcaatg
      481 gcaacgttat atgcagttgg gaatacgaca cagttgtaac tggtccagtt gttacagaca
      541 aaccagaatc ggttgaaatt gatgagccat tattggttcg taaaaagaaa gtacaaaaaa
      601 aggtctctga tgagtaaaag ctcgagggaa tataagtcga cggatccccc gggctgcagg
      661 aatctagacc ttgagtcgaa ttcgagctcg acatatcgtc gctagctgtt ccctttagtg
      721 agggttaatt gcgcgcaatt cccatgtcag ccgttaagtg ttcctgtgtc actcaaaatt
      781 gctttgagag gctctaaggg cttctcagtg cgttacatcc ctggcttgtt gtccacaacc
      841 gttaaacctt aaaagcttta aaagccttat atattctttt ttttcttata aaacttaaaa
      901 ccttagaggc tatttaagtt gctgatttat attaatttta ttgttcaaac atgagagctt
      961 agtacgtgaa acatgagagc ttagtacgtt agccatgaga gcttagtacg ttagccatga
     1021 gggtttagtt cgttaaacat gagagcttag tacgttaaac atgagagctt agtacgtgaa
     1081 acatgagagc ttagtacgta ctatcaacag gttgaactgc tgatcttcag atcctctacg
     1141 ccggacgcat cgtggccgga tcagatctga tatcgtcgca gaccaaaacg atctcaagaa
     1201 gatcatctta ttaatcagat aaaatatttc taggcaccaa taactgcctt aaaaaaatta
     1261 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     1321 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     1381 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
     1441 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc
     1501 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
     1561 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
     1621 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
     1681 gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa gaatgtgaat
     1741 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
     1801 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
     1861 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt
     1921 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     1981 ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa
     2041 gttggcccag ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg
     2101 atcttccgtc acaggtattt attcggcgca attgatatcg cgcgcgtaat acgactcact
     2161 atagggcgaa ttgggtacca gcgcttttcc gctgcataac cctgcttcgg ggtcattata
     2221 gcgatttttt cggtatatcc atcctttttc gcacgatata caggattttg ccaaagggtt
     2281 cgtgtagact ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc
     2341 ccacccgcga gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac
     2401 gggaatcctg ctctgcgagg ctggccggcg tcgacggtat cgataagctt ggcatgcctg
     2461 cagcatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     2521 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agatcgatgc
     2581 ataatgtgcc tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa
     2641 gccgtcaatt gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt
     2701 ttcttcacaa ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg
     2761 cgagaaatag agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg
     2821 ggtggtgctc aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac
     2881 gctaatccct aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg
     2941 ctgtgcgacg ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca
     3001 agcctcgcgt acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg
     3061 ccgcagtaac aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc
     3121 ttgcccggcg ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc
     3181 cgggcgaaag aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta
     3241 ggcgcgcgga cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc
     3301 gtagtgatga atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc
     3361 tcgtccctga tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt
     3421 cattcccagc ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa
     3481 acccgccacc agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc
     3541 agccatactt ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag
     3601 acattgccgt cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct
     3661 tattaaaagc attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag
     3721 tgtctataat cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta
     3781 tgccatagca tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact
     3841 ctctactgtt tctccatacc cgtttttttg g
//
