pK18mobSacB Plasmid

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  • Model: PVT6008
  • 47 Units in Stock
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pK18mobSacB Plasmid

PVT6008  2ug

 

pK18mobSacB Information

Carrier name: pK18mobSacB

Plasmid type: Bacillus subtilis gene knockout carrier

Cloning methods: polyclonal sites, restrictive endonucleases

Carrier size: 5.6kb

5'sequencing primers and sequences: --

3'sequencing primers and sequences: --

Carrier label: -

Carrier resistance: Kanamycin

Use:Homologous recombination HR plasmid

 

pK18mobSacB Description

The Cloning vector allowing mobilization into a wide range of Gramand Gram+ bacteria. After mobilization, the plasmid can be maintained by integration into the host chromosome viahomologous recombination. Excision of the intervening plasmid sequence by a double crossoverevent can be facilitated by selection on medium containing 10% sucrose.The sacB gene has been modified to eliminate the HindIII and EcoRI sites in the coding region.Designation: pK18mobsacB

 

pK18mobSacB Sequence

LOCUS       Exported                5719 bp ds-DNA     circular SYN 13-FEB-2019

DEFINITION  synthetic circular DNA

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 5719)

  AUTHORS   Triple Threat

  TITLE     Direct Submission

FEATURES             Location/Qualifiers

     source          1..5719

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     CDS             346..1140

                     /codon_start=1

                     /gene="aph(3')-II (or nptII)"

                     /product="aminoglycoside phosphotransferase from Tn5"

                     /label=Kan

                     /note="confers resistance to neomycin, kanamycin, and G418 

                     (Geneticin(R))"

                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP

                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS

                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ

                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA

                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"

     promoter        1186..1631

                     /gene="sacR"

                     /label=sacB promoter

                     /note="sacB promoter and control region"

     CDS             1632..3053

                     /codon_start=1

                     /gene="Bacillus subtilis sacB"

                     /product="secreted levansucrase that renders bacterial 

                     growth sensitive to sucrose"

                     /label=SacB

                     /note="negative selection marker"

                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH

                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH

                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW

                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG

                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA

                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE

                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK

                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV

                     VKDSILEQGQLTVNK"

     oriT            3407..3516

                     /note="incP origin of transfer"

     rep_origin      4485..5073

                     /direction=RIGHT

                     /label=ori

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 

                     replication"

BASE COUNT         1538 a         1409 c         1427 g         1344 t

ORIGIN

        1 tgccgcaagc actcagggcg caagggctgc taaaggaagc ggaacacgta gaaagccagt

       61 ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa

      121 aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac

      181 tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag

      241 gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc

      301 aggggatcaa gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat

      361 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca

      421 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg

      481 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactccaaga cgaggcagcg

      541 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact

      601 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct

      661 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg

      721 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt

      781 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc

      841 gcgccagccg aactgttcgc caggctcaag gcgcggatgc ccgacggcga ggatctcgtc

      901 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga

      961 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc

     1021 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt

     1081 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga

     1141 gcgggactct ggggttcgct agaggatcga tcctttttaa cccatcacat atacctgccg

     1201 ttcactatta tttagtgaaa tgagatatta tgatattttc tgaattgtga ttaaaaaggc

     1261 aactttatgc ccatgcaaca gaaactataa aaaatacaga gaatgaaaag aaacagatag

     1321 attttttagt tctttaggcc cgtagtctgc aaatcctttt atgattttct atcaaacaaa

     1381 agaggaaaat agaccagttg caatccaaac gagagtctaa tagaatgagg tcgaaaagta

     1441 aatcgcgcgg gtttgttact gataaagcag gcaagaccta aaatgtgtaa agggcaaagt

     1501 gtatactttg gcgtcacccc ttacatattt taggtctttt tttattgtgc gtaactaact

     1561 tgccatcttc aaacaggagg gctggaagaa gcagaccgct aacacagtac ataaaaaagg

     1621 agacatgaac gatgaacatc aaaaagtttg caaaacaagc aacagtatta acctttacta

     1681 ccgcactgct ggcaggaggc gcaactcaag cgtttgcgaa agaaacgaac caaaagccat

     1741 ataaggaaac atacggcatt tcccatatta cacgccatga tatgctgcaa atccctgaac

     1801 agcaaaaaaa tgaaaaatat caagtttctg aatttgattc gtccacaatt aaaaatatct

     1861 cttctgcaaa aggcctggac gtttgggaca gctggccatt acaaaacgct gacggcactg

     1921 tcgcaaacta tcacggctac cacatcgtct ttgcattagc cggagatcct aaaaatgcgg

     1981 atgacacatc gatttacatg ttctatcaaa aagtcggcga aacttctatt gacagctgga

     2041 aaaacgctgg ccgcgtcttt aaagacagcg acaaattcga tgcaaatgat tctatcctaa

     2101 aagaccaaac acaagaatgg tcaggttcag ccacatttac atctgacgga aaaatccgtt

     2161 tattctacac tgatttctcc ggtaaacatt acggcaaaca aacactgaca actgcacaag

     2221 ttaacgtatc agcatcagac agctctttga acatcaacgg tgtagaggat tataaatcaa

     2281 tctttgacgg tgacggaaaa acgtatcaaa atgtacagca gttcatcgat gaaggcaact

     2341 acagctcagg cgacaaccat acgctgagag atcctcacta cgtagaagat aaaggccaca

     2401 aatacttagt atttgaagca aacactggaa ctgaagatgg ctaccaaggc gaagaatctt

     2461 tatttaacaa agcatactat ggcaaaagca catcattctt ccgtcaagaa agtcaaaaac

     2521 ttctgcaaag cgataaaaaa cgcacggctg agttagcaaa cggcgctctc ggtatgattg

     2581 agctaaacga tgattacaca ctgaaaaaag tgatgaaacc gctgattgca tctaacacag

     2641 taacagatga aattgaacgc gcgaacgtct ttaaaatgaa cggcaaatgg tacctgttca

     2701 ctgactcccg cggatcaaaa atgacgattg acggcattac gtctaacgat atttacatgc

     2761 ttggttatgt ttctaattct ttaactggcc catacaagcc gctgaacaaa actggccttg

     2821 tgttaaaaat ggatcttgat cctaacgatg taacctttac ttactcacac ttcgctgtac

     2881 ctcaagcgaa aggaaacaat gtcgtgatta caagctatat gacaaacaga ggattctacg

     2941 cagacaaaca atcaacgttt gcgccgagct tcctgctgaa catcaaaggc aagaaaacat

     3001 ctgttgtcaa agacagcatc cttgaacaag gacaattaac agttaacaaa taaaaacgca

     3061 aaagaaaatg ccgatgggta ccgagcgaaa tgaccgacca agcgacgccc aacctgccat

     3121 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc

     3181 gggacgccct cgcggacgtg ctcatagtcc acgacgcccg tgattttgta gccctggccg

     3241 acggccagca ggtaggccga caggctcatg ccggccgccg ccgccttttc ctcaatcgct

     3301 cttcgttcgt ctggaaggca gtacaccttg ataggtgggc tgcccttcct ggttggcttg

     3361 gtttcatcag ccatccgctt gccctcatct gttacgccgg cggtagccgg ccagcctcgc

     3421 agagcaggat tcccgttgag caccgccagg tgcgaataag ggacagtgaa gaaggaacac

     3481 ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct gacgccgttg gatacaccaa

     3541 ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat

     3601 accgaaaaaa tcgctataat gaccccgaag cagggttatg cagcggaaaa gcgctgcttc

     3661 cctgctgttt tgtggaatat ctaccgactg gaaacaggca aatgcaggaa attactgaac

     3721 tgaggggaca ggcgagagac gatgccaaag agctcctgaa aatctcgata actcaaaaaa

     3781 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc

     3841 aacgtctcat tttcgccaaa agttggccca gggcttcccg gtatcaacag ggacaccagg

     3901 atttatttat tctgcgaagt gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc

     3961 gggtgatgct gccaacttac tgatttagtg tatgatggtg tttttgaggt gctccagtgg

     4021 cttctgtttc tatcagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat

     4081 cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc tcattttcgc

     4141 caaaagttgg cccagggctt cccggtatca acagggacac caggatttat ttattctgcg

     4201 aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac

     4261 ttactgattt agtgtatgat ggtgtttttg aggtgctcca gtggcttctg tttctatcag

     4321 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaaaagg

     4381 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg

     4441 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt

     4501 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg

     4561 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata

     4621 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca

     4681 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag

     4741 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc

     4801 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga

     4861 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg

     4921 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac

     4981 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg

     5041 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg

     5101 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct

     5161 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc

     5221 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc

     5281 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg

     5341 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta

     5401 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca

     5461 ggaaacagct atgacatgat tacgaattcg agctcggtac ccggggatcc tctagagtcg

     5521 acctgcaggc atgcaagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac

     5581 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat

     5641 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg

     5701 cgataagcta gcttcacgc

//

*Caution

Product is for research use only!
 

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