PEX18GM, 2 ug

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    pEX18GM

    Catalog No. PVT10637
    Packing 2ug

     

    pEX18GM Informaiton

    Replicator: pBR322

    Competent cells: DH5a

    Culture temperature: 37℃

    Host: Escherichia coli

    Uses: Gene knockout

    Prokaryotic resistance: Gen

    Tags: SacB

     

    pEX18GM Description

    PEX18GM is a SacB suicide plasmid of Escherichia coli.
     

    pEX18GM Reference

    1.Biswas I., Mettlach J. (2019) Targeted Gene Replacement in Acinetobacter baumannii. In: Biswas I., Rather P. (eds) Acinetobacter baumannii. Methods in Molecular Biology, vol 1946. Humana Press, New York, NY

    DOI   https://doi.org/10.1007/978-1-4939-9118-1_10
    Publisher Name    Humana Press, New York, NY

     

    pEX18GM Sequence

    LOCUS       Exported                5831 bp ds-DNA     circular SYN 08-JAN-2018

    DEFINITION  synthetic circular DNA

    FEATURES             Location/Qualifiers

         source          1..5831

                         /organism="synthetic DNA construct"

                         /mol_type="other DNA"

         CDS             complement(209..1630)

                         /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

                         ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH

                         IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW

                         SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG

                         KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA

                         YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE

                         IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK

                         MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV

                         VKDSILEQGQLTVNK"

         promoter        complement(1631..2076)

                         /gene="sacR"

                         /label=sacB promoter

                         /note="sacB promoter and control region"

         oriT            2450..2559

                         /note="incP origin of transfer"

         terminator      2980..3007

                         /label=rrnB T2 terminator

                         /note="transcription terminator T2 from the E. coli rrnB 

                         gene"

         terminator      3099..3185

                         /gene="Escherichia coli rrnB"

                         /label=rrnB T1 terminator

                         /note="transcription terminator T1 from the E. coli rrnB 

                         gene"

         CDS             complement(3341..3625)

                         /codon_start=1

                         /gene="lacZ fragment"

                         /product="LacZ-alpha fragment of beta-galactosidase"

                         /label=lacZ-alpha

                         /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR

                         LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRAWLFWRMREDFQPDTD"

         misc_feature    3555..3611

                         /label=MCS

                         /note="pUC18/19 multiple cloning site"

         protein_bind    3645..3661

                         /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)."

         promoter        complement(3669..3699)

                         /label=lac promoter

                         /note="promoter for the E. coli lac operon"

         protein_bind    3714..3735

                         /label=CAP binding site

                         /bound_moiety="E. coli catabolite activator protein"

                         /note="CAP binding activates transcription in the presence 

                         of cAMP."

         rep_origin      complement(4023..4611)

                         /direction=LEFT

                         /label=ori

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

                         replication"

         CDS             complement(4864..5397)

                         /codon_start=1

                         /gene="aacC1"

                         /product="gentamycin acetyltransferase"

                         /label=Gen

                         /note="confers resistance to gentamycin"

                         /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD

                         LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS

                         EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR

                         EEVMHFDIDPSTAT"

         promoter        complement(5586..5614)

                         /gene="intI1 (promoter lies within the coding sequence)"

                         /label=Pc promoter

                         /note="class 1 integron promoter"

         promoter        complement(5632..5736)

                         /gene="bla"

                         /label=AmpR promoter

    ORIGIN

            1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca

           61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg

          121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc

          181 accataatcg gcattttctt ttgcgttttt atttgttaac tgttaattgt ccttgttcaa

          241 ggatgctgtc tttgacaaca gatgttttct tgcctttgat gttcagcagg aagctaggcg

          301 caaacgttga ttgtttgtct gcgtagaatc ctctgtttgt catatagctt gtaatcacga

          361 cattgtttcc tttcgcttga ggtacagcga agtgtgagta agtaaaggtt acatcgttag

          421 gatcaagatc catttttaac acaaggccag ttttgttcag cggcttgtat gggccagtta

          481 aagaattaga aacataacca agcatgtaaa tatcgttaga cgtaatgccg tcaatcgtca

          541 tttttgatcc gcgggagtca gtgaacagat accatttgcc gttcatttta aagacgttcg

          601 cgcgttcaat ttcatctgtt actgtgttag atgcaatcag cggtttcatc acttttttca

          661 gtgtgtaatc atcgtttagc tcaatcatac cgagagcgcc gtttgctaac tcagccgtgc

          721 gttttttatc gctttgcaga agtttttgac tttcttgacg gaagaatgat gtgcttttgc

          781 catagtatgc tttgttaaat aaagattctt cgccttggta gccatcttca gttccagtgt

          841 ttgcttcaaa tactaagtat ttgtggcctt tatcttctac gtagtgagga tctctcagcg

          901 tatggttgtc gcctgagctg tagttgcctt catcgatgaa ctgctgtaca ttttgatacg

          961 tttttccgtc accgtcaaag attgatttat aatcctctac accgttgatg ttcaaagagc

         1021 tgtctgatgc tgatacgtta acttgtgcag ttgtcagtgt ttgtttgccg taatgtttac

         1081 cggagaaatc agtgtagaat aaacggattt ttccgtcaga tgtaaatgtg gctgaacctg

         1141 accattcttg tgtttggtct tttaggatag aatcatttgc atcgaatttg tcgctgtctt

         1201 taaagacgcg gccagcgttt ttccagctgt caatagaagt ttcgccgact ttttgataga

         1261 acatgtaaat cgatgtgtca tccgcatttt taggatctcc ggctaatgca aagacgatgt

         1321 ggtagccgtg atagtttgcg acagtgccgt cagcgttttg taatggccag ctgtcccaaa

         1381 cgtccaggcc ttttgcagaa gagatatttt taattgtgga cgaatcgaac tcaggaactt

         1441 gatatttttc atttttttgc tgttcaggga tttgcagcat atcatggcgt gtaatatggg

         1501 aaatgccgta tgtttcctta tatggctttt ggttcgtttc tttcgcaaac gcttgagttg

         1561 cgcctcctgc cagcagtgcg gtagtaaagg ttaatactgt tgcttgtttt gcaaactttt

         1621 tgatgttcat cgttcatgtc tcctttttta tgtactgtgt tagcggtctg cttcttccag

         1681 ccctcctgtt tgaagatggc aagttagtta cgcacaataa aaaaagacct aaaatatgta

         1741 aggggtgacg ccaaagtata cactttgccc tttacacatt ttaggtcttg cctgctttat

         1801 cagtaacaaa cccgcgcgat ttacttttcg acctcattct attagactct cgtttggatt

         1861 gcaactggtc tattttcctc ttttgtttga tagaaaatca taaaaggatt tgcagactac

         1921 gggcctaaag aactaaaaaa tctatctgtt tcttttcatt ctctgtattt tttatagttt

         1981 ctgttgcatg ggcataaagt tgccttttta atcacaattc agaaaatatc ataatatctc

         2041 atttcactaa ataatagtga acggcaggta tatgtgatgg gttaaaaagg atcgatcctc

         2101 tagctagagt cgatcttcgc cagcagggcg aggatcgtgg catcaccgaa ccgcgccgtg

         2161 cgcgggtcgt cggtgagcca gagtttcagc aggccgccca ggcggcccag gtcgccattg

         2221 atgcgggcca gctcgcggac gtgctcatag tccacgacgc ccgtgatttt gtagccctgg

         2281 ccgacggcca gcaggtaggc cgacaggctc atgccggccg ccgccgcctt ttcctcaatc

         2341 gctcttcgtt cgtctggaag gcagtacacc ttgataggtg ggctgccctt cctggttggc

         2401 ttggtttcat cagccatccg cttgccctca tctgttacgc cggcggtagc cggccagcct

         2461 cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa

         2521 cacccgctcg cgggtgggcc tacttcacct atcctgcccg gctgacgccg ttggatacac

         2581 caaggaaagt ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga

         2641 tataccgaaa aaatcgctat aatgaccccg aagcagggtt atgcagcgga aaagcgctgc

         2701 ttccctgctg ttttgtggaa tatctaccga ctggaaacag gcaaatgcag gaaattactg

         2761 aactgagggg acaggcgaga gacgatgcca aagagctaca ccgacgagct ggccgagtgg

         2821 gttgaatccc gcgcggccaa gaagcgccgg cgtgatgagg ctgcggttgc gttcctggcg

         2881 gtgagggcgg atgtcgatat gcgtaaggag aaaataccgc atcaggcgca tatttgaatg

         2941 tatttagaaa aataaacaaa aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg

         3001 gccttctgct taatttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg

         3061 ggccgttgct tcgcaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg

         3121 ttcaccgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt

         3181 atttgatgcc tggcagttcc ctactctcgc atggggagac cccacactac catcggcgct

         3241 acggcgtttc acttctgagt tcggcatggg gtcaggtggg accaccgcgc tactgccgcc

         3301 aggcaaattc tgttttatca gaccgcttct gcgttctgat ttaatctgta tcaggctgaa

         3361 aatcttctct catccgccaa aacagccaag ctcgccattc gccattcagg ctgcgcaact

         3421 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat

         3481 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa

         3541 cgacggccag tgccaagctt gcatgcctgc aggtcgactc tagaggatcc ccgggtaccg

         3601 agctcgaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca

         3661 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg

         3721 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg

         3781 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc

         3841 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta

         3901 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag

         3961 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg

         4021 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg

         4081 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg

         4141 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga

         4201 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc

         4261 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt

         4321 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact

         4381 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg

         4441 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt

         4501 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt

         4561 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct

         4621 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg

         4681 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt

         4741 aaatcaatct aaagtatata tgagtaaact tggtctgaca atcgatgcga attggccgcg

         4801 gcgttgtgac aatttaccga acaactccgc ggccgggaag ccgatctcgg cttgaacgaa

         4861 ttgttaggtg gcggtacttg ggtcgatatc aaagtgcatc acttcttccc gtatgcccaa

         4921 ctttgtatag agagccactg cgggatcgtc accgtaatct gcttgcacgt agatcacata

         4981 agcaccaagc gcgttggcct catgcttgag gagattgatg agcgcggtgg caatgccctg

         5041 cctccggtgc tcgccggaga ctgcgagatc atagatatag atctcactac gcggctgctc

         5101 aaacctgggc agaacgtaag ccgcgagagc gccaacaacc gcttcttggt cgaaggcagc

         5161 aagcgcgatg aatgtcttac tacggagcaa gttcccgagg taatcggagt ccggctgatg

         5221 ttgggagtag gtggctacgt ctccgaactc acgaccgaaa agatcaagag cagcccgcat

         5281 ggatttgact tggtcagggc cgagcctaca tgtgcgaatg atgcccatac ttgagccacc

         5341 taactttgtt ttagggcgac tgccctgctg cgtaacatcg ttgctgctgc gtaacatcgt

         5401 tgctgctcca taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc

         5461 gaggcataga ctgtacaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt

         5521 taccaccgct gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc

         5581 attacagttt acgaaccgaa caggcttatg tcaattcggt tgaatactca tactcttcct

         5641 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga

         5701 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc

         5761 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag

         5821 gccctttcgt c

    //

    Caution:
    1.  This product is FOR RESEARCH USE ONLY!
    2.  The item is lyophilized form, Please take the powder plasmid by centrifugation at 5000rpm/min for 1min. Add 20μl ddH2O in to the tube of plasmid.