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ID   PSM23      preliminary; circular DNA; SYN; 7689 BP.
XX
AC   ATCC68179;
XX
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE   Bacillus plasmid vector pSM23 - complete.
XX
KW   cloning vector.
XX
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
XX
RN   [1]
RC   pSM1 from pE194 & pBR322
RC   pSM3 from pSM1
RC   pSM6, pSM7, pSM8 from pSM3 & linker
RC   pSM100 from pUB110
RC   pSM17, pSM23 from pSM100 & pSM6 & pSM7
RC   pSM26 from pSM23 & linker
RC   pSM27 from pSM17
RC   pSM29 from pSM23
RC   pSM112 from pSM29
RC   pSM117 from pSM29
RC   pSM115 from pSM112 & pBR322
RC   pSM119 from pSM115 & pSM112
RC   pSM134 from pSM112 & pLV59
RC   pSM135 from pSM134 & pSM112
RA   Grandi G., Del Bue M., Palla E., Mele A., Colletti E., Toma S.;
RT   "New plasmid expression vectors for Bacillus subtilis";
RL   Plasmid 16:1-14(1986).
XX
CC   Created by Moore, July 1995, under contract with NCBI.
CC   Restriction digests of the clone give the following sizes (kb):
CC   XbaI--uncut; EcoRI--uncut; BamHI--7.6. (ATCC staff)
CC   Medium is -1 1065 plus kanamycin (5 ug/ml).
CC   NM (pSM23)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (Bacillus subtilus BD170)(Bacillus subtilis)
CC   CP ()
CC   FN (cloning)
CC   SE ()
CC   PA (pUB110)(pE194)
CC   BR ()
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pE194 PstI 3728bp 232..232
FT                   2. pBR322 PstI 4361bp 3612..3612
FT                   -> pSM1 8089bp [tet in E. coli, ery in B.subtilis]
FT                   1. pSM1 EcoRI 8089bp, pBR322 4360..4360
FT                   DNA polymerase I with AT
FT                   -> pSM3 8080bp [no EcoRI]
FT                   1. pSM3 large HpaI-SstI/SacI 7320bp,
FT                   \ remove pE194 2142..2901
FT                   EcoRI linker 6bp gaattc:
FT                   2. pE194 HaeIII 3728bp 2614..2614, 120bp 3' to ery RBS
FT                   exonuclease III
FT                   S1 nuclease, remove 120 bp from ends
FT                   SstI/SacI-blunt 160bp,
FT                   \ 156bp 5' to RBS/10bp 3' to ery RBS
FT                   -> pSM6 7480bp [EcoRI 4bp 3' to RBS]
FT                   1. pSM3 large HpaI-SstI/SacI 7320bp,
FT                   \ remove pE194 2142..2901
FT                   EcoRI linker 6bp gaattc:
FT                   2. pE194 HaeIII 3728bp 2614..2614, 120bp 3' to ery RBS
FT                   exonuclease III
FT                   S1 nuclease, remove 120 bp from ends
FT                   SstI/SacI-blunt 160bp,
FT                   \ 156bp 5' to RBS/10bp 3' to ery RBS
FT                   -> pSM7 7000bp [EcoRI starts in RBS]
FT                   1. pUB110 EcoRI 4548bp 2..2
FT                   DNA polymerase I with AT
FT                   -> pSM100 4500bp
FT                   1. pSM100 XbaI 4500bp, pUB110 465..465
FT                   2. pSM7 remove PstI-PstI 3400bp,
FT                   \ pE194 232..232/pBR322 3614..3614/3600bp
FT                   XbaI 3600bp, pE194 3480..3480
FT                   -> pSM23 7600bp"
FT   -               1..5
FT                   /note="pUB110 1..5 5bp
FT                   EcoRI = G^AATT C
FT                   EcoRI =      G^AATTC"
FT   -               6..472
FT                   /note="pUB110 2..468 467bp
FT                   XbaI = T^CTAG A
FT                   PstI =      C TGCA^G"
FT   -               473..2386
FT                   /note="pE194 228..2141 1914bp
FT                   HpaI = GTT^AAC
FT                   \              ttataaccctctt..."
FT   -               2387..3605
FT                   /note="pE194 2741..3728..231 1219bp
FT                   PstI = CTGCA^G
FT                   XbaI =     T^CTAGA"
FT   -               3606..7689
FT                   /note="pUB110 465..4548 4084bp"
FT   rep_origin      0..0
FT                   /note="ORI Bacillus pUB110"
FT   CDS             0..0
FT                   /note="ANT Bacillus kanamycin resistance gene (kan)"
XX
SQ   Sequence 7689 BP; 2487 A; 1379 C; 1272 G; 2551 T; 0 other;
     gaattaattc cttagtgctt tcatagatta aactcacatc acgctttaaa tcgcttattt
     tagactttaa agacttgttt tcttcaagca actcattata atcatttaca ttttcattaa
     atcgctctac aagaccacta tatttttctt taacttgccc atgttcttta cttaattttt
     tatattctct cgccatatca gtactcatga gatttctaac atgctgtttt aacctatcgt
     tatctctcgc agcagtcact aagtttttat aatcacgctc cgatataaca acatttttgg
     ttggtttctt ttctgttttc attatttctt ttcccaaacc aaacatggac ttttcacccg
     ttggcacttc aacacttttc atgtgtcgtt tcgctggtac ttctaaatct gatttaactt
     tatcgctata agcagtccat tcatcttttt taactgctaa atttttttct agtgcagaac
     ggattgttga tgattacgaa aatattaaga gcacagacta ttacacagaa aatcaagaat
     taaaaaaacg tagagagagt ttgaaagaag tagtgaatac atggaaagag gggtatcacg
     aaaaaagtaa agaggttaat aaattaaagc gagagaatga tagtttgaat gagcagttga
     atgtatcaga gaaatttcaa gctagtacag tgactttata tcgtgctgcg agggcgaatt
     tccctgggtt tgagaaaggg tttaataggc ttaaagagaa attctttaat gattccaaat
     ttgagcgtgt gggacagttt atggatgttg tacaggataa tgtccagaag gtcgatagaa
     agcgtgagaa acagcgtaca gacgatttag agatgtagag gtacttttat gccgagaaaa
     ctttttgcgt gtgacagtcc ttaaaatata cttagagcgt aagcgaaagt agtagcgaca
     gctattaact ttcggttgca aagctctagg atttttaatg gacgcagcgc atcacacgca
     aaaaggaaat tggaataaat gcgaaatttg agatgttaat taaagacctt tttgaggtct
     ttttttctta gatttttggg gttatttagg ggagaaaaca taggggggta ctacgacctc
     ccccctaggt gtccattgtc cattgtccaa acaaataaat aaatattggg tttttaatgt
     taaaaggttg ttttttatgt taaagtgaaa aaaacagatg ttgggaggta cagtgatggt
     tgtagataga aaagaagaga aaaaagttgc tgttacttta agacttacaa cagaagaaaa
     tgagatatta aatagaatca aagaaaaata taatattagc aaatcagatg caaccggtat
     tctaataaaa aaatatgcaa aggaggaata cggtgcattt taaacaaaaa aagatagaca
     gcactggcat gctgcctatc tatgactaaa ttttgttaag tgtattagca ccgttattat
     atcatgagcg aaaatgtaat aaaagaaact gaaaacaaga aaaattcaag aggacgtaat
     tggacatttg ttttatatcc agaatcagca aaagccgagt ggttagagta tttaaaagag
     ttacacattc aatttgtagt gtctccatta catgataggg atactgatac agaaggtagg
     atgaaaaaag agcattatca tattctagtg atgtatgagg gtaataaatc ttatgaacag
     ataaaaataa ttaacagaag aattgaatgc gactattccg cagattgcag gaagtgtgaa
     aggtcttgtg agatatatgc ttcacatgga cgatcctaat aaatttaaat atcaaaaaga
     agatatgata gtttatggcg gtgtagatgt tgatgaatta ttaaagaaaa caacaacaga
     tagatataaa ttaattaaag aaatgattga gtttattgat gaacaaggaa tcgtagaatt
     taagagttta atggattatg caatgaagtt taaatttgat gattggttcc cgcttttatg
     tgataactcg gcgtatgtta ttcaagaata tataaaatca aatcggtata aatctgaccg
     atagattttg aatttaggtg tcacaagaca ctcttttttc gcaccagcga aaactggttt
     aagccgactg cgcaaaagac ataatcgatt cacaaaaaat aggcacacga aaaacaagtt
     aagggatgca gtttatgcat cccttaactt acttattaaa taatttatag ctattgaaaa
     gagataagaa ttgttcaaag ctaatattgt ttaaatcgtc aattcctgca tgttttaagg
     aattgttaaa ttgatttttt gtaaatattt tcttgtattc tttgtttaac cctctttaat
     ttggttatat gaattttgct tattaacgat tcattataac cacttatttt ttgtttggtt
     gataatgaac tgtgctgatt acaaaaatac taaaaatgcc catatttttt cctccttata
     aaattagtat aattatagca cgagctctga taaatatgaa catgatgagt gatcgttaaa
     tttatactgc aatcggatgc gattattgaa taaaagatat gagagattta tctaatttct
     tttttcttgt aaaaaaagaa agttcttaaa ggttttatag ttttggtcgt agagcacacg
     gtttaacgac ttaattacga agtaaataag tctagtgtgt tagactttat gaaatctata
     tacgtttata tatatttatt atccggaggt gtagcatgtc tcattcaatt ttgagggttg
     ccagagttaa aggatcaagt aatacaaacg ggatacaaag acataatcaa agagagaata
     aaaactataa taataaagac ataaatcatg aggaaacata taaaaattat gatttgatta
     acgcacaaaa tataaagtat aaagataaaa ttgatgaaac gattgatgag aattattcag
     ggaaacgtaa aattcggtca gatgcaattc gacatgtgga cggactggtt acaagtgata
     aagatttctt tgatgattta agcggagaag aaatagaacg attttttaaa gatagcttgg
     agtttctaga aaatgaatac ggtaaggaaa atatgctgta tgcgactgtc catctggatg
     aaagagtccc acatatgcac tttggttttg tccctttaac agaggacggg agattgtctg
     caaaagaaca gttaggcaac aagaaagact ttactcaatt acaagataga tttaatgagt
     atgtgaatga gaaaggttat gaacttgaaa gaggcacgtc caaagaggtt acagaacgag
     aacataaagc gatggatcag tacaagaaag atactgtatt tcataaacag gaactgcaag
     aagttaagga tgagttacag aaggcaaata agcagttaca gagtggaata gagcatatga
     ggtctacgaa accctttgat tatgaaaatg agcgtacagg tttgttctct ggacgtgaag
     agactggtag aaagatatta actgctgatg aatttgaacg cctgcaagaa acaatctctt
     ctgcactaga aaatcaatct ctttttccaa agtttgtttt ttaaatttag ctgtctcaat
     atgtttacgg tcagagccac gttcaccacg cttcaactca aaaccctgtt ttttcatatg
     ctcggggaat ttatcttgta gccataacag ttcttgacga ttaaacacat tttttccttg
     cagttttcca tcacgcatag gcacaacacc taaatgcatg tgaggggttt gctcatcatt
     atgaactgtt gcataagcaa tattttgctt gccatatcgt tcggaaaata atttataact
     ttcctcaaaa aatcgttttt gttctcctgg atccagttgc tcaaaaaaat ctcggtcaga
     tgttactagc aactcattta caagaacagc atctttcctc cgtttttctt gtacctgttt
     tttgtgattc aataatttct ttgacacgtt cgttgtaatc aatattttta tcatttttca
     aatcataatt ttcacgtgtt cgctcatggt caatatcatc attcgttcta ctttttcgct
     ctctttgatt atgaaattgc atgcctttta gtccagctga tttcactttt tgcattctac
     aaactgcata actcatatgt aaatcgctcc tttttaggtg gcacaaatgt gaggcatttt
     cgctctttcc ggcaaccact tccaagtaaa gtataacaca ctatacttta tattcataaa
     gtgtgtgctc tgcgaggctg tcggcagtgc cgaccaaaac cataaaacct ttaagacctt
     tctttttttt acgagaaaaa agaaacaaaa aaacctgccc tctgccacct cagcaaaggg
     gggttttgct ctcgtgctcg tttaaaaatc agcaagggac aggtagtatt ttttgagaag
     atcactcaaa aaatctccac ctttaaaccc ttgccaattt ttattttgtc cgttttgtct
     agcttaccga aagccagact cagcaagaat aaaattttta ttgtctttcg gttttctagt
     gtaacggaca aaaccactca aaataaaaaa gatacaagag aggtctctcg tatcttttat
     tcagcaatcg cgcccgattg ctgaacagat taataataga ttttagcttt ttatttgttg
     aaaaaagcta atcaaattgt tgtcgggatc aattactgca aagtctcgtt catcccacca
     ctgatctttt aatgatgtat tggggtgcaa aatgcccaaa ggcttaatat gttgatataa
     ttcatcaatt ccctctactt caatgcggca actagcagta ccagcaataa acgactccgc
     acctgtacaa accggtgaat cattactacg agagcgccag cttcatcact tgcctcccat
     agatgaatcc gaacctcatt acacattaga actgcgaatc catcttcatg gtgaaccaaa
     gtgaaaccta gtttatcgca ataaaaacct atactctttt taatatcccc gactggcaat
     gccgggatag actgtaacat tctcacgcat aaaatcccct ttcattttct aatgtaaatc
     tattacctta ttattaattc aattcgctca taattaatcc tttttcttat tacgcaaaat
     ggcccgattt aagcacaccc tttattccgt taatgcgcca tgacagccat gataattact
     aatactagga gaagttaata aatacgtaac caacatgatt aacaattatt agaggtcatc
     gttcaaaatg gtatgcgttt tgacacatcc actatatatc cgtgtcgttc tgtccactcc
     tgaatcccat tccagaaatt ctctagcgat tccagaagtt tctcagagtc ggaaagttga
     ccagacatta cgaactggca cagatggtca taacctgaag gaagatctga ttgcttaact
     gcttcagtta agaccgaagc gctcgtcgta taacagatgc gatgatgcag accaatcaac
     atggcacctg ccattgctac ctgtacagtc aaggatggta gaaatgttgt cggtccttgc
     acacgaatat tacgccattt gcctgcatat tcaaacagct cttctacgat aagggcacaa
     atcgcatcgt ggaacgtttg ggcttctacc gatttagcag tttgatacac tttctctaag
     tatccacctg aatcataaat cggcaaaata gagaaaaatt gaccatgtgt aagcggccaa
     tctgattcca cctgagatgc ataatctagt agaatctctt cgctatcaaa attcacttcc
     accttccact caccggttgt ccattcatgg ctgaactctg cttcctctgt tgacatgaca
     cacatcatct caatatccga atagggccca tcagtctgac gaccaagaga gccataaaca
     ccaatagcct taacatcatc cccatattta tccaatattc gttccttaat ttcatgaaca
     atcttcattc tttcttctct agtcattatt attggtccat tcactattct cattcccttt
     tcagataatt ttagatttgc ttttctaaat aagaatattt ggagagcacc gttcttattc
     agctattaat aactcgtctt cctaagcatc cttcaatcct tttaataaca attatagcat
     ctaatcttca acaaactggc ccgtttgttg aactactctt taataaaata atttttccgt
     tcccaattcc acattgcaat aatagaaaat ccatcttcat cggctttttc gtcatcatct
     gtatgaatca aatcgccttc ttctgtgtca tcaaggttta attttttatg tatttctttt
     aacaaaccac cataggagat taacctttta cggtgtaaac cttcctccaa atcagacaaa
     cgtttcaaat tcttttcttc atcatcggtc ataaaatccg tatcctttac aggatatttt
     gcagtttcgt caattgccga ttgtatatcc gatttatatt tatttttcgg tcgaatcatt
     tgaactttta catttggatc atagtctaat ttcattgcct ttttccaaaa ttgaatccat
     tgtttttgat tcacgtagtt ttctgtattc ttaaaataag ttggttccac acataccaat
     acatgcatgt gctgattata agaattatct ttattattta ttgtcacttc cgttgcacgc
     ataaaaccaa caagattttt attaattttt ttatattgca tcattcggcg aaatccttga
     gccatatctg acaaactctt atttaattct tcgccatcat aaacattttt aactgttaat
     gtgagaaaca accaacgaac tgttggcttt tgtttaataa cttcagcaac aaccttttgt
     gactgaatgc catgtttcat tgctctcctc cagttgcaca ttggacaaag cctggattta
     caaaaccaca ctcgatacaa ctttctttcg cctgtttcac gattttgttt atactctaat
     atttcagcac aatcttttac tctttcagcc tttttaaatt caagaatatg cagaagttca
     aagtaatcaa cattagcgat tttcttttct ctccatggtc tcacttttcc actttttgtc
     ttgtccacta aaacccttga tttttcatct gaataaatgc tactattagg acacataata
     ttaaaagaaa cccccatcta tttagttatt tgtttagtca cttataactt taacagatgg
     ggtttttctg tgcaaccaat tttaagggtt ttcaatactt taaaacacat acataccaac
     acttcaacgc acctttcagc aactaaaata aaaatgacgt tatttctata tgtatcaaga
     taagaaagaa caagttcaaa accatcaaaa aaagacacct tttcaggtgc tttttttatt
     ttataaactc attccctgat ctcgacttcg ttcttttttt acctctcggt tatgagttag
     ttcaaattcg ttctttttag gttctaaatc gtgtttttct tggaattgtg ctgttttatc
     ctttaccttg tctacaaacc ccttaaaaac gtttttaaag gcttttaagc cgtctgtacg
     ttccttaag
//