pDS132

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  • Model: PVT10634
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pDS132

Catalog No. PVT10634
Packing 2ug
 

pDS132 Informaiton

Prokaryotic resistance: chloramphenicol Chl

Screening markers: SacB

Cloned strain: Escherichia coli S17-1 PIR

Culture conditions: 37 degrees centigrade

 

pDS132 Description

PDS132 is a SacB suicide plasmid of Escherichia coli

 

pDS132 Sequence

LOCUS       Exported                5286 bp ds-DNA     circular SYN 08-JAN-2018

DEFINITION  synthetic circular DNA

FEATURES             Location/Qualifiers

     source          1..5286

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     rep_origin      4..392

                     /label=R6K gamma ori

                     /note="gamma replication origin from E. coli plasmid R6K; 

                     requires the R6K initiator protein pi for replication"

     CDS             complement(1010..1381)

                     /codon_start=1

                     /gene="traJ"

                     /product="oriT-recognizing protein"

                     /label=traJ

                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV

                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE

                     KQDELGKVMMGVVRPRAEP"

     oriT            1414..1523

                     /note="incP origin of transfer"

     CDS             complement(2489..3148)

                     /codon_start=1

                     /gene="cat"

                     /product="chloramphenicol acetyltransferase"

                     /label=Chl

                     /note="confers resistance to chloramphenicol"

                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL

                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS

                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM

                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"

     CDS             complement(3246..4667)

                     /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(4668..5113)

                     /gene="sacR"

                     /label=sacB promoter

                     /note="sacB promoter and control region"

     protein_bind    5239..5255

                     /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)."

ORIGIN

        1 ccatgtcagc cgttaagtgt tcctgtgtca ctcaaaattg ctttgagagg ctctaagggc

       61 ttctcagtgc gttacatccc tggcttgttg tccacaaccg ttaaacctta aaagctttaa

      121 aagccttata tattcttttt tttcttataa aacttaaaac cttagaggct atttaagttg

      181 ctgatttata ttaattttat tgttcaaaca tgagagctta gtacgtgaaa catgagagct

      241 tagtacgtta gccatgagag cttagtacgt tagccatgag ggtttagttc gttaaacatg

      301 agagcttagt acgttaaaca tgagagctta gtacgtgaaa catgagagct tagtacgtac

      361 tatcaacagg ttgaactgct gatcttcaga tcctctacgc cggacgcatc gtggccggat

      421 ccagccgacc aggctttcca cgcccgcgtg ccgctccatg tcgttcgcgc ggttctcgga

      481 aacgcgctgc cgcgtttcgt gattgtcacg ctcaagcccg tagtcccgtt cgagcgtcgc

      541 gcagaggtca gcgagggcgc ggtaggcccg atacggctca tggatggtgt ttcgggtcgg

      601 gtgaatcttg ttgatggcga tatggatgtg caggttgtcg gtgtcgtgat gcacggcact

      661 gacgcgctga tgctcggcga agccaagccc agcgcagatg cggtcctcaa tcgcgcgcaa

      721 cgtctccgcg tcgggcttct ctcccgcgcg gaagctaacc agcaggtgat aggtcttgtc

      781 ggcctcggaa cgggtgttgc cgtgctgggt cgccatcacc tcggccatga cagcgggcag

      841 ggtgtttgcc tcgcagttcg tgacgcgcac gtgacccagg cgctcggtct tgccttgctc

      901 gtcggtgatg tacttcacca gctccgcgaa gtcgctcttc ttgatggagc gcatggggac

      961 gtgcttggca atcacgcgca ccccccggcc gttttagcgg ctaaaaaagt catggctctg

     1021 ccctcgggcg gaccacgccc atcatgacct tgccaagctc gtcctgcttc tcttcgatct

     1081 tcgccagcag ggcgaggatc gtggcatcac cgaaccgcgc cgtgcgcggg tcgtcggtga

     1141 gccagagttt cagcaggccg cccaggcggc ccaggtcgcc attgatgcgg gccagctcgc

     1201 ggacgtgctc atagtccacg acgcccgtga ttttgtagcc ctggccgacg gccagcaggt

     1261 aggccgacag gctcatgccg gccgccgccg ccttttcctc aatcgctctt cgttcgtctg

     1321 gaaggcagta caccttgata ggtgggctgc ccttcctggt tggcttggtt tcatcagcca

     1381 tccgcttgcc ctcatctgtt acgccggcgg tagccggcca gcctcgcaga gcaggattcc

     1441 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg

     1501 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac

     1561 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg

     1621 ctataatgac cccgaagcag ggttatgcag cggaaaagcg ctgcttccct gctgttttgt

     1681 ggaatatcta ccgactggaa acaggcaaat gcaggaaatt actgaactga ggggacaggc

     1741 gagagacgat gccaaagagc tacaccgacg agctggccga gtgggttgaa tcccgcgcgg

     1801 ccaagaagcg ccggcgtgat gaggctgcgg ttgcgttcct ggcggtgagg gcggatgtcg

     1861 aggcggcgtt agcgtccggc tatgcgctcg tcaccatttg ggagcacatg cgggaaacgg

     1921 ggaaggtcaa gttctcctac gagacgttcc gctcgcacgc caggcggcac atcaaggcca

     1981 agcccgccga tgtgcccgca ccgcaggcca aggctgcgga acccgcgccg gcacccaaga

     2041 cgccggagcc acggcggccg aagcaggggg gcaaggctga aaagccggcc cccgctgcgg

     2101 ccccgaccgg cttcaccttc aacccaacac cggacaaaaa ggatcctcta cgccggacgc

     2161 atcgtggccg gcatcaccgg cgccacaagt gcggttgctg gcgcctatat cgccgacatc

     2221 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt

     2281 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgct gcctcgcgcg

     2341 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg

     2401 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg

     2461 gtgtcggggc gcagccatga cccgggaatt acgccccgcc ctgccactca tcgcagtact

     2521 gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct

     2581 gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa

     2641 cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc

     2701 agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt

     2761 tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt

     2821 ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag

     2881 ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgg aattccggat

     2941 gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt

     3001 tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt

     3061 gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg

     3121 tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgcc ctatgggatt

     3181 cacctttatg ttgataagaa ataaaagaaa atgccaatag gatatcggca ttttcttttg

     3241 cgtttttatt tgttaactgt taattgtcct tgttcaagga tgctgtcttt gacaacagat

     3301 gttttcttgc ctttgatgtt cagcaggaag cttggcgcaa acgttgattg tttgtctgcg

     3361 tagaatcctc tgtttgtcat atagcttgta atcacgacat tgtttccttt cgcttgaggt

     3421 acagcgaagt gtgagtaagt aaaggttaca tcgttaggat caagatccat ttttaacaca

     3481 aggccagttt tgttcagcgg cttgtatggg ccagttaaag aattagaaac ataaccaagc

     3541 atgtaaatat cgttagacgt aatgccgtca atcgtcattt ttgatccgcg ggagtcagtg

     3601 aacaggtacc atttgccgtt cattttaaag acgttcgcgc gttcaatttc atctgttact

     3661 gtgttagatg caatcagcgg tttcatcact tttttcagtg tgtaatcatc gtttagctca

     3721 atcataccga gagcgccgtt tgctaactca gccgtgcgtt ttttatcgct ttgcagaagt

     3781 ttttgacttt cttgacggaa gaatgatgtg cttttgccat agtatgcttt gttaaataaa

     3841 gattcttcgc cttggtagcc atcttcagtt ccagtgtttg cttcaaatac taagtatttg

     3901 tggcctttat cttctacgta gtgaggatct ctcagcgtat ggttgtcgcc tgagctgtag

     3961 ttgccttcat cgatgaactg ctgtacattt tgatacgttt ttccgtcacc gtcaaagatt

     4021 gatttataat cctctacacc gttgatgttc aaagagctgt ctgatgctga tacgttaact

     4081 tgtgcagttg tcagtgtttg tttgccgtaa tgtttaccgg agaaatcagt gtagaataaa

     4141 cggatttttc cgtcagatgt aaatgtggct gaacctgacc attcttgtgt ttggtctttt

     4201 aggatagaat catttgcatc gaatttgtcg ctgtctttaa agacgcggcc agcgtttttc

     4261 cagctgtcaa tagaagtttc gccgactttt tgatagaaca tgtaaatcga tgtgtcatcc

     4321 gcatttttag gatctccggc taatgcaaag acgatgtggt agccgtgata gtttgcgaca

     4381 gtgccgtcag cgttttgtaa tggccagctg tcccaaacgt ccaggccttt tgcagaagag

     4441 atatttttaa ttgtggacga atcgaattca ggaacttgat atttttcatt tttttgctgt

     4501 tcagggattt gcagcatatc atggcgtgta atatgggaaa tgccgtatgt ttccttatat

     4561 ggcttttggt tcgtttcttt cgcaaacgct tgagttgcgc ctcctgccag cagtgcggta

     4621 gtaaaggtta atactgttgc ttgttttgca aactttttga tgttcatcgt tcatgtctcc

     4681 ttttttatgt actgtgttag cggtctgctt cttccagccc tcctgtttga agatggcaag

     4741 ttagttacgc acaataaaaa aagacctaaa atatgtaagg ggtgacgcca aagtatacac

     4801 tttgcccttt acacatttta ggtcttgcct gctttatcag taacaaaccc gcgcgattta

     4861 cttttcgacc tcattctatt agactctcgt ttggattgca actggtctat tttcctcttt

     4921 tgtttgatag aaaatcataa aaggatttgc agactacggg cctaaagaac taaaaaatct

     4981 atctgtttct tttcattctc tgtatttttt atagtttctg ttgcatgggc ataaagttgc

     5041 ctttttaatc acaattcaga aaatatcata atatctcatt tcactaaata atagtgaacg

     5101 gcaggtatat gtgatgggtt aaaaaggatc gatcctctag agtcgacctg caggtcgacg

     5161 gatctgcagg tcgacggatc ccaagcttct tctagaggta ccgcatgcga tatcgagctc

     5221 tcccgggaat tccacaaatt gttatccgct cacaattcca catgtggaat tccacatgtg

     5281 gaattc

//

 

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