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.